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Planta Med 2018; 84(16): 1183-1190
DOI: 10.1055/a-0626-7426
Biological and Pharmacological Activity
Original Papers
Georg Thieme Verlag KG Stuttgart · New York

Anti-inflammatory Effects of Cardamonin in Ovarian Cancer Cells Are Mediated via mTOR Suppression

Huajiao Chen
Department of Pharmacy, Fujian Provincial Maternity and Childrenʼs Hospital, Affiliated Hospital of Fujian Medical University, Fuzhou, Fujian, China
,
Daohua Shi
Department of Pharmacy, Fujian Provincial Maternity and Childrenʼs Hospital, Affiliated Hospital of Fujian Medical University, Fuzhou, Fujian, China
,
Peiguang Niu
Department of Pharmacy, Fujian Provincial Maternity and Childrenʼs Hospital, Affiliated Hospital of Fujian Medical University, Fuzhou, Fujian, China
,
Yanting Zhu
Department of Pharmacy, Fujian Provincial Maternity and Childrenʼs Hospital, Affiliated Hospital of Fujian Medical University, Fuzhou, Fujian, China
,
Jintuo Zhou
Department of Pharmacy, Fujian Provincial Maternity and Childrenʼs Hospital, Affiliated Hospital of Fujian Medical University, Fuzhou, Fujian, China
› Author Affiliations
Further Information

Correspondence

Prof. Dr. Daohua Shi
Department of Pharmacy
Fujian Provincial Maternity and Childrenʼs Hospital
18 Daoshan Road, Fuzhou
Fujian, 350001
China   
Phone: + 86 5 91 88 31 20 79   
Fax: + 86 5 91 88 31 20 79   

Publication History

received 27 December 2017
revised 19 April 2018

accepted 30 April 2018

Publication Date:
17 May 2018 (online)

Also available at
 
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Abstract

Cardamonin exhibits a variety of pharmacological activities including anti-inflammatory and antitumor, which are correlated with the inhibition of nuclear factor-kappaB and the mammalian target of rapamycin, respectively. However, whether the anti-inflammatory effects of cardamonin are mediated by the mammalian target of rapamycin remains unknown. In this study, ovarian cancer SKOV3 cells were cultured with lipopolysaccharide to induce inflammation, and the inhibitory effects and underlying molecular mechanisms of cardamonin were investigated using specific inhibitors of the mammalian target of rapamycin and the nuclear factor-kappaB pathway (rapamycin and pyrrolidine dithiocarbamate, respectively). Our results indicated that cardamonin inhibited the viability of normal and lipopolysaccharide-pretreated SKOV3 cells in a concentration-dependent manner. In accordance with rapamycin, the activation of the mammalian target of rapamycin and its downstream target, ribosomal protein S6 kinase 1, was inhibited by cardamonin, while pyrrolidine dithiocarbamate substantially blocked nuclear factor-kappaB activation and mildly inhibited the phosphorylation of the mammalian target of rapamycin and ribosomal protein S6 kinase 1. Pretreated with pyrrolidine dithiocarbamate, the effect of cardamonin on the mammalian target of rapamycin signalling was not affected, but the expression of inflammatory factors was further reduced. In cells pretreated with rapamycin, the inhibitory effects of cardamonin were completely suppressed with regards to the phosphorylation of the mammalian target of rapamycin, ribosomal protein S6 kinase 1, TNF-α, and interleukin-6, and nuclear factor-kappaB p65 protein expression was decreased. In conclusion, our findings indicate that the anti-inflammatory effects of cardamonin are correlated with mammalian target of rapamycin inhibition.


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Key words

cardamonin - inflammation - lipopolysaccharide - mTOR - NF-κB - ovarian cancer

Abbreviations

Akt: protein kinase B
IKK: inhibitor of NF-κB kinase
IL: interleukin
LPS: lipopolysaccharide
mTOR: mammalian target of rapamycin
mTORC1: mammalian target of rapamycin complex 1
NF-κB: nuclear factor-kappaB
PDTC: pyrrolidine dithiocarbamate
PI3K: phosphatidylinositol 3 kinase
PTEN: phosphate and tension homology
Raptor: regulatory associated protein of mTOR
S6K1: ribosomal protein S6 kinase 1
TNF-α : tumor necrosis factor α
 

Introduction

Ovarian cancer is a highly aggressive neoplasm with a low 5-year survival rate that seriously threatens womenʼs health worldwide [1], [2]. Extensive studies have shown that inflammation is closely associated with the growth, invasion, and angiogenesis of ovarian cancer, thus anti-inflammatory treatments are becoming a new strategy for ovarian cancer therapy [3].

The mTOR, a highly conserved serine/threonine kinase, is critical for cancer initiation and progression. It represents a promising target for cancer treatment [4]. Previous studies have revealed that mTOR regulates the expression of inflammatory factors, such as IL-6, and transcription factors, such as NF-κB, which lead to inflammation [5], [6]. Moreover, recent studies have shown that mTORC1 is an important signalling intermediary in inflammation-mediated tumor progression [7], [8]. On the other hand, the downregulation of mTOR with a specific mTOR inhibitor leads to the inhibition of inflammatory cytokine expression and NF-κB phosphorylation [9], [10]. This appears to be an important mechanism by which mTOR inhibitors regulate the proliferation of cancer cells [11], [12], [13]. Collectively, these findings demonstrate that the activity of mTOR is closely related to tumor inflammation, and therefore regulating mTOR activity may be an effective target in cancer treatment.

Cardamonin is a natural chalcone derived from the seeds of Alpinia katsumadai Hayat. It has anti-inflammatory, antioxidation, antitumor, vasodilator, and other pharmacological activities [14] ([Fig. 1]). Our previous studies have revealed that cardamonin inhibits hypoxia-induced tumor angiogenesis, and cellular proliferation of Lewis lung carcinoma cells, vascular smooth muscle cells, and ovarian cancer SKOV3 cells [15], [16]. In addition, we also found that the regulation of cardamonin in glucose metabolism was related to mTOR inhibition [17], [18]. Other studies have shown that cardamonin exhibits anti-inflammatory activity via NF-κB inhibition [19], [20]. However, whether the anti-inflammatory effects of cardamonin are mediated by mTOR remains unknown.

Zoom Image
Fig. 1 The chemical structure of cardamonin.

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Results

Our results indicated that cardamonin inhibited the cell viability of LPS-treated and non-treated cells in a concentration-dependent manner. As expected, rapamycin (0.1 µM) also decreased the cell viability of both treated and non-treated cells ([Fig. 2]).

Zoom Image
Fig. 2 Cardamonin effects on the cell viability of normal and LPS-pretreated SKOV3 cells. Cells were pretreated with or without LPS (1 µg/mL) for 30 min, and then treated with cardamonin (1, 3, 10, 30, and 100 µM) and rapamycin (0.1 µM) for 48 h, respectively. Cell viability was then measured by the MTT assay (n = 5). Data are presented as the mean ± SEM of five experiments. *P < 0.05, **p < 0.01 compared with the control group; #p < 0.05, ##p < 0.01 compared with the LPS-pretreated group.

Cardamonin inhibited the expression of inflammatory factors in SKOV3 cells. To investigate the effects of cardamonin on inflammation, we analyzed the cellular secretion of inflammatory factors. As shown in [Fig. 3], cardamonin (3 and 30 µM) and rapamycin (0.1 µM) inhibited the protein expression of TNF-α and IL-6 under both conditions. Neither cardamonin nor rapamycin had a significant effect on the expression of IL-8.

Zoom Image
Fig. 3 Cardamonin inhibited the expression of TNF-α and IL-6. SKOV3 cells were pretreated with or without LPS (1 µg/mL) for 30 min, and then treated with cardamonin (3 or 30 µM) and rapamycin (0.1 µM) for 24 h, respectively. Secreted concentrations of TNF-α, IL-6, and IL-8 in the media were determined by ELISA (n = 5). A Concentration of TNF-α. B Concentration of IL-6. C Concentration of IL-8. Data are presented by the mean ± SEM of five experiments. **P < 0.01 compared with the control group; ##p < 0.01 compared with the LPS-pretreated group.

Cardamonin also inhibited nuclear NF-κB protein expression in SKOV3 cells. NF-κB is closely related to the initiation, progression, and metastasis of inflammation-associated tumors. Cardamonin inhibited NF-κB activity in a concentration-dependent manner by decreasing the level of nuclear NF-κB ([Fig. 4]).

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Fig. 4 Cardamonin decreased the activation of NF-κB p65. SKOV3 cells were pretreated with or without LPS (1 µg/mL) for 30 min, and then treated with cardamonin (3 or 30 µM) and rapamycin (0.1 µM) for 24 h, respectively. Total protein and nuclear proteins were extracted for Western blot analysis (n = 3). A The protein bands of total NF-κB p65, nuclear NF-κB p65, and histone H2A. B The relative density ratio of nuclear NF-κB p65 was normalized to histone H2A. Data are presented as the mean ± SEM of three experiments. **P < 0.01 compared with the control group; ##p < 0.01 compared with the LPS-pretreated group.

The anti-inflammatory effects of cardamonin were found to be associated with NF-κB and mTOR. mTOR plays an important role in the regulation of cancer and inflammation. In this study, we attempted to investigate the effects of cardamonin on mTOR under mimicked inflammatory conditions. LPS induced the expression of inflammatory factors and NF-κB, and, at the same time, we detected a significant increase in the expression of p-mTOR and p-S6K1. Treated with cardamonin (3 and 30 µM) and rapamycin (0.1 µM), the phosphorylation of mTOR and S6K1 were sharply reduced, while a negligible effect on the total protein expression of mTOR and S6K1 was observed ([Fig. 5]).

Zoom Image
Fig. 5 Cardamonin decreased the activation of mTOR and S6K1. SKOV3 cells were pretreated with or without LPS (1 µg/mL) for 30 min, and then treated with cardamonin (3 or 30 µM) and rapamycin (0.1 µM) for 24 h, respectively. Total protein was extracted for Western blot analysis (n = 3). A The protein bands of mTOR, p-mTOR, S6K1, p-S6K1, Akt, p-Akt, and actin. B The relative density ratios of p-mTOR, p-S6K1, and p-Akt protein were normalized to actin. Data are presented as the mean ± SEM of three experiments. *P < 0.05, **p < 0.01 compared with the control group; #p < 0.05, ##p < 0.01 compared with the LPS-pretreated group.

We then examined the effects of cardamonin on Akt, which is upstream of NF-κB and mTOR. As expected, LPS stimulated the phosphorylation of Akt. Rapamycin and a low concentration of cardamonin (3 µM) had no significant effect on the expression of p-Akt under both conditions. However, the expression of p-Akt was increased with the higher concentration of cardamonin (30 µM) in normal SKOV3 cells, but no effect was observed with LPS treatment ([Fig. 5]). These findings suggested that the anti-inflammatory effects of cardamonin may be associated with NF-κB and mTOR, but not with Akt.

To furtherly investigate the anti-inflammatory mechanisms of cardamonin, mTOR and NF-κB were blocked by rapamycin and PDTC, respectively. In the PDTC (100 µM) group, the expression of inflammatory factors, including TNF-α and IL-6, was remarkably reduced; no significant effect on IL-8 expression was observed. In cells pretreated with PDTC and LPS, the expression of TNF-α and IL-6 was reduced further after cardamonin treatment ([Fig. 6]). In addition, PDTC mildly suppressed the phosphorylation of mTOR and S6K1 that was induced by LPS. In cells pretreated with PDTC, the inhibitory effects of cardamonin on NF-κB were blocked, but no effect on mTOR signalling was observed. In cells pretreated with rapamycin, the suppressive activity of cardamonin on mTOR and inflammatory factors disappeared, but the expression of nuclear NF-κB p65 was slightly weakened ([Figs. 7] and [8]). These results indicated that the anti-inflammatory effects of cardamonin were correlated with mTOR inhibition.

Zoom Image
Fig. 6 Effects of cardamonin on the expression of TNF-α, IL-6, and IL-8 in mTOR and NF-κB inhibitor pretreated cells. Cells were cultured with rapamycin (0.1 µM) and PDTC (100 µM) for 1 h, respectively, and then treated with LPS (1 µg/mL) for 30 min; lastly, cells were treated with cardamonin (30 µM) for 24 h. Secreted concentrations of TNF-α, IL-6, and IL-8 in the media were determined by ELISA (n = 5). A Concentration of TNF-α. B Concentration of IL-6. C Concentration of IL-8. Data are presented as the mean ± SEM of five experiments. **P < 0.01 compared with the control group; ##p < 0.01 compared with the LPS-pretreated group; &&p < 0.05 compared with the LPS + cardamonin group.
Zoom Image
Fig. 7 Effects of cardamonin on the activation of NF-κB p65 in mTOR and NF-κB inhibitor pretreated cells. Cells were cultured with rapamycin (0.1 µM) and PDTC (100 µM) for 1 h, respectively, and then treated with LPS (1 µg/mL) for 30 min; lastly, cells were treated with cardamonin (30 µM) for 24 h. Total protein and nuclear proteins were extracted for Western blot analysis (n = 3). A The protein bands of total NF-κB p65, nuclear NF-κB p65, and histone H2A. B The relative density ratio of nuclear NF-κB p65 was normalized to histone H2A. Data are presented as the mean ± SEM of three experiments. **P < 0.01 compared with the control group; ##p < 0.01 compared with the LPS-pretreated group; &&p < 0.05 compared with the LPS + cardamonin group.
Zoom Image
Fig. 8 Effects of cardamonin on the activation of mTOR and S6K1 in mTOR and NF-κB inhibitor pretreated cells. Cells were cultured with rapamycin (0.1 µM) and PDTC (100 µM) for 1 h, respectively, and then treated with LPS (1 µg/mL) for 30 min; lastly, cells were treated with cardamonin (30 µM) for 24 h. Total protein was extracted for Western blot analysis (n = 3). A The protein bands of mTOR, p-mTOR, S6K1, p-S6K1, Akt, p-Akt, and actin. B The relative density ratios of p-mTOR, p-S6K1, and p-Akt proteins were normalized to actin. Data are presented as the mean ± SEM of three experiments. **P < 0.01 compared with the control group; #p < 0.05, ##p < 0.01 compared with the LPS-pretreated group; &&p < 0.05 compared with the LPS + cardamonin group.

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Discussion

Increasing evidence has shown that inflammation plays a pivotal role in the initiation and development of various cancers [21], [22]. Therefore, studies focused on inflammation in ovarian cancer aim to reveal the cancer pathogenesis and provide a new strategy for treatment.

LPS, the endotoxic part of the gram-negative bacteria, is often used to mimic inflammatory conditions in laboratory experiments. LPS combines with Toll-like receptor 4, which then activates the PI3K/Akt pathway [23], [24], promoting the production of inflammatory cytokines and inducing unremitting inflammatory activity. In this study, our results showed higher levels of inflammatory cytokines, including TNF-α, IL-6, and IL-8, and nuclear proteins, such as NF-κB p65, when cells were incubated with LPS, indicating that inflammatory conditions were successfully created.

Recent progress has shown that mTOR partakes in the progression of inflammation-mediated tumors as a key modulator [8], [25]. Indeed, mTOR-dependent hyperproliferation of the colonic epithelium leads to inflammation-associated tumorigenesis in mice with inflammatory bowel disease [26]. Accordingly, rapamycin and its analogues suppress mTOR activity and have been shown to hinder the course of inflammation-related tumors [11], [25]. Therefore, these previous studies have confirmed an association between inflammation-mediated tumors and mTOR activation. In our present study, the activation of mTOR and its substrate, S6K1, which was induced by LPS, was inhibited by cardamonin, indicating cardamonin may be a potential mTOR inhibitor. In addition, the inhibitory effects of cardamonin on the expression of TNF-α and IL-6 suggest an underlying relation between cardamonin and inflammation.

Extensive studies have demonstrated that mTOR regulates NF-κB activation [27], [28], which promotes the expression of inflammatory cytokines [5]. In PTEN inactive prostate cancer cells, the Raptor regulates NF-κB activity by activating the endogenous IKK, which is upstream of NF-κB [29]. Rapamycin suppresses TLR2-induced inflammatory responses through the downregulation of NF-κB signalling [30]. Moreover, it has been reported that rapamycin suppresses not only NF-κB phosphorylation but also NF-κB nuclear transcription in colon cancer cells [11]. This inhibitory mechanism may be explained by the fact that rapamycin dissociates Raptor from mTORC1 and thereby inhibits Raptor from interacting with IKK, which ultimately results in the decreased expression of IKK and NF-κB [29]. In this study, we found that cardamonin decreased the phosphorylation of mTOR as well as the expression of NF-κB p65 nuclear protein. In addition, our previous study discovered that cardamonin decreased both the expression and phosphorylation of Raptor, indicating cardamonin may be a specific mTORC1 inhibitor [31]. Therefore, we speculate that the anti-inflammatory effects of cardamonin involve the downregulation of NF-κB activity via the mTOR pathway.

To further investigate the anti-inflammatory effects of cardamonin, we blocked mTOR and NF-κB with rapamycin and PDTC, respectively, in LPS pretreated SKOV3 cells. It has been reported that PDTC impedes the transfer of NF-κB into the cell nucleus [32]. As expected, PDTC remarkably downregulated the expression of nuclear NF-κB p65 as well as the inflammatory factors TNF-α and IL-6. In PDTC pretreated cells, the inhibitory effects of cardamonin on NF-κB were blocked, while the expression of mTOR signalling, including that of TNF-α and IL-6, was slightly decreased. However, when cells were pretreated with rapamycin, the suppressive activity of cardamonin on mTOR and the inflammatory factors were almost completely blocked, while the expression of nuclear NF-κB p65 was slightly decreased. These results suggested that the anti-inflammatory effects of cardamonin were correlated with mTOR inhibition. In addition, the discordant suppression of NF-κB and the inflammatory cytokines may be explained by the fact that not only NF-κB, but also mTOR, can directly regulate inflammation [33]. Surprisingly, we also found that PDTC mildly reduced the phosphorylation of mTOR and S6K1, suggesting that there may be a feedback loop between mTOR and NF-κB. Coincidently, previous studies have shown that activated NF-κB leads to elevated mTOR expression, and mTOR may be suppressed by NF-κB inhibition [34].

Akt is downstream of PI3K and upstream of NF-κB and mTOR. When sensing mitogens and cytokines, Akt phosphorylates downstream targets and facilitates various biological effects [35]. Because our research has identified that the anti-inflammatory effects of cardamonin are likely associated with NF-κB and mTOR, we also wanted to explore the effect of cardamonin on the expression of Akt. As expected, LPS phosphorylated Akt, leading to its activation. Low doses (3 µM) of cardamonin and rapamycin had no significant effect on the expression of p-Akt in non-treated cells or those treated with LPS, while a high dose (30 µM) of cardamonin only increased the expression of p-Akt in normal SKOV3 cells. This phenomenon may be due to a negative feedback loop [36]. We speculate that the intensive inhibition of 30 µM cardamonin on mTOR leads to the activation of Akt, while LPS mimicked inflammation and increased mTOR activation, thus depressing the negative feedback loop. Consistent with our results, previous studies have demonstrated that a high activation of mTORC1 negatively regulates the PI3K/Akt signalling pathway, while inhibiting mTORC1 disrupts this feedback, resulting in the mTORC2-dependent activation of Akt [37].

Inflammatory factors play important roles in the development of ovarian cancer [38], [39]. In this study, cardamonin and rapamycin inhibited the expression of TNF-α and IL-6 under both conditions. This finding is consistent with those of previous studies wherein cardamonin has been reported to exhibit anti-inflammatory effects [40]. Further studies have demonstrated that cardamonin inhibits inflammation through p65NF-kappaB inhibition in LPS-stimulated RAW 264.7 macrophages and BV2 microglia [20], [41]. The anti-inflammatory effect of cardamonin had been demonstrated in a concentration range from 2 to 50 µM [41], so 3 and 30 µM of cardamonin were used in the present study. Interestingly, both TNF-α and IL-6 appear to be sensitive to the low dose of cardamonin under normal conditions, as opposed to the high dose of cardamonin in LPS-mimicked inflammatory conditions. This may be explained by the fact that a high concentration of cardamonin leads to negative feedback in order to activate Akt, which upregulates the expression of TNF-α and IL-6. However, similar to the NF-κB inhibition by PDTC, neither cardamonin nor rapamycin had a significant effect on the expression of IL-8. Similarly, studies have demonstrated that cardamonin inhibits the nuclear transduction of NF-κB and further reduces the secretion of IL-6 and CCL5, but not IL-8 [42]. Other studies have shown that NF-κB inhibition is ineffective at blocking cytokine-induced IL-8 production, while P38 and other signal transducers as well as activators of transcription (STAT1) inhibitors were effective [43]. In contrast to the studies above, Lin et al. found that mTOR inhibition downregulated the expression of chemokines in monocytes, including IL-8 [44]. In consideration of cell type and drug treatment time, we hypothesized that there may be other signalling pathways, aside from mTOR/NF-κB, that could regulate the secretion of IL-8 in SKOV3 cells.

These findings suggest that cardamonin inhibits LPS-induced inflammation, and this effect is correlated with mTOR inhibition. Thus, cardamonin was considered as a potential agent for the treatment of inflammatory-related tumors.


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Materials and Methods

Materials

Solutions and supplements for cell cultures were purchased from Gibco. Cardamonin (NO. 110763, purity > 99%) was purchased from the National Institutes for Food and Drug Control of China. Rapamycin, LPS, PDTC, and MTT were from Sigma. Antibodies against Akt, p-Akt (Ser473), mTOR, p-mTOR (Ser2448), S6K1, and p-S6K1 (Thr389) were obtained from Cell Signalling Technology. Antibodies against actin, NF-κBp65, and histone H2A as well as the secondary antibody (anti-rabbit IgG, mouse radish peroxidase-linked antibody) were from Santa Cruz Biotechnology. The primers were designed and purchased from the Shanghai Shenggong Bioengineering Institute.


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Cell culture

SKOV3 cells were purchased from Boster Biological Technology Co., Ltd. and cultured in McCoyʼs 5 A medium with 10% FBS, 100 U/mL penicillin, and 100 µg/mL streptomycin at 37 °C in an atmosphere containing 5% CO2. SKOV3 cells were cultured with LPS (1 µg/mL) to mimic inflammatory conditions.


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Cell viability assays

Cell viability was determined by the MTT assay. Cells were grown in 96-well plates and then treated with different drugs for 48 h (n = 5). MTT (0.5 mg/mL, 20 µL) was added to each well, and cells were incubated at 37 °C for 4 h. After discarding the supernatant carefully, 150 µL DMSO were added to each well. Each 96-well plate was shaken for 15 min, and the absorbance was determined at 490 nm by a microplate reader (Model 1680, Bio-Rad).


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Inflammatory factors measurement

SKOV3 cells were treated with indicated drugs for 24 h. Culture supernatants were collected and clarified by centrifugation before the evaluation of cytokine production using ELISA. Concentrations of TNF-α, IL-6, and IL-8 were measured using a commercial ELISA kit from R&D Systems following the manufacturerʼs instructions. Results were normalized to the cell count.


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Western blotting

Protein expression assays were performed by Western blot. After drug exposure, cells were washed twice with ice-cold PBS and resuspended in lysis buffer. Lysates were centrifuged at 14 000 × g at 4 °C for 20 min, and the resultant supernatant was used for experiments. The solubilized protein content was determined by the BCA method, and 30 µg of proteins were resolved by SDS-PAGE and subsequently electrotransferred to nitrocellulose membranes for conventional immunoblotting. After blocking in 5% bovine serum albumin, they were incubated overnight with the primary antibody. Antibodies against actin, Akt, p-Akt (Ser473), mTOR, p-mTOR (Ser2448), S6K1, and p-S6K1 (Thr389) were diluted to 1 : 1000, and antibodies against NF-κBp65 and histone H2A were diluted to 1 : 500. Finally, membranes were incubated with horseradish peroxidase-coupled goat anti-rabbit antibody (1 : 2000) reacted with the HRP-ECL chemiluminescence reagent for 1 – 3 min, followed by exposure to X-ray film in order to produce bands within the linear range.


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Statistical analysis

Statistical analysis was performed using SPSS 16.0 software. All data are expressed as the mean ± SEM. Differences between groups were evaluated by the Studentʼs t-test or one-way ANOVA followed by the Dunnettʼs post hoc test. A p value less than 0.05 was considered statistically significant.


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Conflict of Interest

The authors declare no conflicts of interest.

Acknowledgements

This work was supported by the Youth Scientific Foundation and Innovative Medical Foundation of Fujian Provincial Health and Family Planning Commission [2014 – 2 – 6 and 2016-CX-13] and the Natural Science Foundation of Fujian Province [2016J01492 and 2017J01234], China.

  • References

  • 1 Chen T, Jansen L, Gondos A, Emrich K, Holleczek B, Katalinic A, Luttmann S, Meyer M, Brenner H. GEKID Cancer Survival Working Group. Survival of ovarian cancer patients in Germany in the early 21st century: a period analysis by age, histology, laterality, and stage. Eur J Cancer Prev 2013; 22: 59-67
    CrossrefPubMedSearch in Google Scholar
  • 2 Edwards HM, Noer MC, Sperling CD, Nguyen-Nielsen M, Lundvall L, Christensen IJ, Hogdall C. Survival of ovarian cancer patients in Denmark: Results from the Danish gynaecological cancer group (DGCG) database, 1995–2012. Acta Oncol 2016; 55 (Suppl. 02) 36-43
    CrossrefPubMedSearch in Google Scholar
  • 3 Xie X, Yang M, Ding Y, Chen J. Microbial infection, inflammation and epithelial ovarian cancer. Oncol Lett 2017; 14: 1911-1919
    CrossrefPubMedSearch in Google Scholar
  • 4 Villar VH, Nguyen TL, Teres S, Bodineau C, Duran RV. Escaping mTOR inhibition for cancer therapy: Tumor suppressor functions of mTOR. Mol Cell Oncol 2017; 4: e1297284
    PubMedSearch in Google Scholar
  • 5 Vangan N, Cao Y, Jia X, Bao W, Wang Y, He Q, Binderiya U, Feng X, Li T, Hao H, Wang Z. mTORC1 mediates peptidoglycan induced inflammatory cytokines expression and NF-κB activation in macrophages. Microb Pathog 2016; 99: 111-118
    CrossrefPubMedSearch in Google Scholar
  • 6 Temiz-Resitoglu M, Kucukkavruk SP, Guden DS, Cecen P, Sari AN, Tunctan B, Gorur A, Tamer-Gumus L, Buharalioglu CK, Malik KU, Sahan-Firat S. Activation of mTOR/IκB-α/NF-κB pathway contributes to LPS-induced hypotension and inflammation in rats. Eur J Pharmacol 2017; 802: 7-19
    CrossrefPubMedSearch in Google Scholar
  • 7 Lee DF, Kuo HP, Chen CT, Hsu JM, Chou CK, Wei Y, Sun HL, Li LY, Ping B, Huang WC, He X, Hung JY, Lai CC, Ding Q, Su JL, Yang JY, Sahin AA, Hortobagyi GN, Tsai FJ, Tsai CH, Hung MC. IKK beta suppression of TSC1 links inflammation and tumor angiogenesis via the mTOR pathway. Cell 2007; 130: 440-455
    CrossrefPubMedSearch in Google Scholar
  • 8 Dufour M, Faes S, Dormond-Meuwly A, Demartines N, Dormond O. PGE2-induced colon cancer growth is mediated by mTORC1. Biochem Biophys Res Comm 2014; 451: 587-591
    CrossrefPubMedSearch in Google Scholar
  • 9 Pan H, Xu LH, Ouyang DY, Wang Y, Zha QB, Hou XF, He XH. The second-generation mTOR kinase inhibitor INK128 exhibits anti-inflammatory activity in lipopolysaccharide-activated RAW 264.7 cells. Inflammation 2014; 37: 756-765
    CrossrefPubMedSearch in Google Scholar
  • 10 Bao W, Wang Y, Fu Y, Jia X, Li J, Vangan N, Bao L, Hao H, Wang Z. mTORC1 regulates flagellin-induced inflammatory response in macrophages. PLoS One 2015; 10: e0125910
    CrossrefPubMedSearch in Google Scholar
  • 11 Sun Q, Liu Q, Zheng Y, Cao X. Rapamycin suppresses TLR4-triggered IL-6 and PGE(2) production of colon cancer cells by inhibiting TLR4 expression and NF-kappaB activation. Mol Immunol 2008; 45: 2929-2936
    CrossrefPubMedSearch in Google Scholar
  • 12 Chandrika G, Natesh K, Ranade D, Chugh A, Shastry P. Suppression of the invasive potential of Glioblastoma cells by mTOR inhibitors involves modulation of NFkappaB and PKC-alpha signaling. Sci Rep 2016; 6: 22455
    CrossrefPubMedSearch in Google Scholar
  • 13 Ekshyyan O, Khandelwal AR, Rong X, Moore-Medlin T, Ma X, Alexander JS, Nathan CO. Rapamycin targets interleukin 6 (IL-6) expression and suppresses endothelial cell invasion stimulated by tumor cells. Am J Transl Res 2016; 8: 4822-4830
    PubMedSearch in Google Scholar
  • 14 Goncalves LM, Valente IM, Rodrigues JA. An overview on cardamonin. J Med Food 2014; 17: 633-640
    CrossrefPubMedSearch in Google Scholar
  • 15 Niu PG, Zhang YX, Shi DH, Liu Y, Chen YY, Deng J. Cardamonin inhibits metastasis of Lewis lung carcinoma cells by decreasing mTOR activity. PLoS One 2015; 10: e0127778
    CrossrefPubMedSearch in Google Scholar
  • 16 Xue ZG, Niu PG, Shi DH, Liu Y, Deng J, Chen YY. Cardamonin inhibits angiogenesis by mTOR downregulation in SKOV3 cells. Planta Med 2016; 82: 70-75
    Thieme ConnectPubMedSearch in Google Scholar
  • 17 Liao Q, Shi DH, Zheng W, Xu XJ, Yu YH. Antiproliferation of cardamonin is involved in mTOR on aortic smooth muscle cells in high fructose-induced insulin resistance rats. Eur J Pharmacol 2010; 641: 179-186
    CrossrefPubMedSearch in Google Scholar
  • 18 Niu P, Zhang Y, Shi D, Chen Y, Deng J. Cardamonin ameliorates insulin resistance induced by high insulin and high glucose through the mTOR and signal pathway. Planta Med 2013; 79: 452-458
    Thieme ConnectPubMedSearch in Google Scholar
  • 19 Lee JH, Jung HS, Giang PM, Jin X, Lee S, Son PT, Lee D, Hong YS, Lee K, Lee JJ. Blockade of nuclear factor-kappaB signaling pathway and anti-inflammatory activity of cardamomin, a chalcone analog from Alpinia conchigera . J Pharmacol Exp Ther 2006; 316: 271-278
    CrossrefPubMedSearch in Google Scholar
  • 20 Chow YL, Lee KH, Vidyadaran S, Lajis NH, Akhtar MN, Israf DA, Syahida A. Cardamonin from Alpinia rafflesiana inhibits inflammatory responses in IFN-gamma/LPS-stimulated BV2 microglia via NF-kappaB signalling pathway. Int Immunopharmacol 2012; 12: 657-665
    CrossrefPubMedSearch in Google Scholar
  • 21 Shan W, Liu J. Inflammation: a hidden path to breaking the spell of ovarian cancer. Cell Cycle 2009; 8: 3107-3111
    CrossrefPubMedSearch in Google Scholar
  • 22 White KL, Schildkraut JM, Palmieri RT, Iversen jr. ES, Berchuck A, Vierkant RA, Rider DN, Charbonneau B, Cicek MS, Sutphen R, Birrer MJ, Pharoah PP, Song H, Tyrer J, Gayther SA, Ramus SJ, Wentzensen N, Yang HP, Garcia-Closas M, Phelan CM, Cunningham JM, Fridley BL, Sellers TA, Goode EL. Ovarian Cancer Association Consortium. Ovarian cancer risk associated with inherited inflammation-related variants. Cancer Res 2012; 72: 1064-1069
    CrossrefPubMedSearch in Google Scholar
  • 23 Cho JS, Kang JH, Um JY, Han IH, Park IH, Lee HM. Lipopolysaccharide induces pro-inflammatory cytokines and MMP production via TLR4 in nasal polyp-derived fibroblast and organ culture. PLoS One 2014; 9: e90683
    CrossrefPubMedSearch in Google Scholar
  • 24 Laird MH, Rhee SH, Perkins DJ, Medvedev AE, Piao W, Fenton MJ, Vogel SN. TLR4/MyD88/PI3K interactions regulate TLR4 signaling. J Leukoc Biol 2009; 85: 966-977
    CrossrefPubMedSearch in Google Scholar
  • 25 Thiem S, Pierce TP, Palmieri M, Putoczki TL, Buchert M, Preaudet A, Farid RO, Love C, Catimel B, Lei Z, Rozen S, Gopalakrishnan V, Schaper F, Hallek M, Boussioutas A, Tan P, Jarnicki A, Ernst M. mTORC1 inhibition restricts inflammation-associated gastrointestinal tumorigenesis in mice. J Clin Invest 2013; 123: 767-781
    PubMedSearch in Google Scholar
  • 26 Deng L, Zhou JF, Sellers RS, Li JF, Nguyen AV, Wang Y, Orlofsky A, Liu Q, Hume DA, Pollard JW, Augenlicht L, Lin EY. A novel mouse model of inflammatory bowel disease links mammalian target of rapamycin-dependent hyperproliferation of colonic epithelium to inflammation-associated tumorigenesis. Am J Pathol 2010; 176: 952-967
    CrossrefPubMedSearch in Google Scholar
  • 27 Li Z, Zhang J, Mulholland M, Zhang W. mTOR activation protects liver from ischemia/reperfusion-induced injury through NF-kappaB pathway. FASEB J 2017; 31: 3018-3026
    CrossrefPubMedSearch in Google Scholar
  • 28 Wang Y, Zhang X, Tang W, Lin Z, Xu L, Dong R, Li Y, Li J, Zhang Z, Li X, Zhao L, Wei JJ, Shao C, Kong B, Liu Z. miR-130a upregulates mTOR pathway by targeting TSC1 and is transactivated by NF-kappaB in high-grade serous ovarian carcinoma. Cell Death Differ 2017; 24: 2089-2100
    CrossrefPubMedSearch in Google Scholar
  • 29 Dan HC, Cooper MJ, Cogswell PC, Duncan JA, Ting JP, Baldwin AS. Akt-dependent regulation of NF-{kappa}B is controlled by mTOR and Raptor in association with IKK. Genes Dev 2008; 22: 1490-1500
    CrossrefPubMedSearch in Google Scholar
  • 30 Sun R, Zhang Y, Ma S, Qi H, Wang M, Duan J, Ma S, Zhu X, Li G, Wang H. Down-regulation of mitogen-activated protein kinases and nuclear factor-kappaB signaling is involved in rapamycin suppression of TLR2-induced inflammatory response in monocytic THP-1 cells. Microbiol Immunol 2015; 59: 614-622
    CrossrefPubMedSearch in Google Scholar
  • 31 Shi D, Zhu Y, Niu P, Zhou J, Chen H. Raptor mediates the antiproliferation of cardamonin by mTORC1 inhibition in SKOV3 cells. Onco Targets Ther 2018; 11: 757-767
    CrossrefPubMedSearch in Google Scholar
  • 32 Morais C, Pat B, Gobe G, Johnson DW, Healy H. Pyrrolidine dithiocarbamate exerts anti-proliferative and pro-apoptotic effects in renal cell carcinoma cell lines. Nephrol Dial Transplant 2006; 21: 3377-3388
    CrossrefPubMedSearch in Google Scholar
  • 33 Wu D, Cheng J, Sun G, Wu S, Li M, Gao Z, Zhai S, Li P, Su D, Wang X. p70S6K promotes IL-6-induced epithelial-mesenchymal transition and metastasis of head and neck squamous cell carcinoma. Oncotarget 2016; 7: 36539-36550
    CrossrefPubMedSearch in Google Scholar
  • 34 Okamoto T, Ozawa Y, Kamoshita M, Osada H, Toda E, Kurihara T, Nagai N, Umezawa K, Tsubota K. The neuroprotective effect of rapamycin as a modulator of the mTOR-NF-kappaB axis during retinal inflammation. PLoS One 2016; 11: e0146517
    CrossrefPubMedSearch in Google Scholar
  • 35 Wendel HG, De Stanchina E, Fridman JS, Malina A, Ray S, Kogan S, Cordon-Cardo C, Pelletier J, Lowe SW. Survival signalling by Akt and eIF4E in oncogenesis and cancer therapy. Nature 2004; 428: 332-337
    PubMedSearch in Google Scholar
  • 36 Park S, Zhao D, Hatanpaa KJ, Mickey BE, Saha D, Boothman DA, Story MD, Wong ET, Burma S, Georgescu MM, Rangnekar VM, Chauncey SS, Habib AA. RIP1 activates PI3K-Akt via a dual mechanism involving NF-kappaB-mediated inhibition of the mTOR-S6K-IRS1 negative feedback loop and down-regulation of PTEN. Cancer Res 2009; 69: 4107-4111
    CrossrefPubMedSearch in Google Scholar
  • 37 Carracedo A, Ma L, Teruya-Feldstein J, Rojo F, Salmena L, Alimonti A, Egia A, Sasaki AT, Thomas G, Kozma SC, Papa A, Nardella C, Cantley LC, Baselga J, Pandolfi PP. Inhibition of mTORC1 leads to MAPK pathway activation through a PI3K-dependent feedback loop in human cancer. J Clin Invest 2008; 118: 3065-3074
    PubMedSearch in Google Scholar
  • 38 Lane D, Matte I, Garde-Granger P, Laplante C, Carignan A, Rancourt C, Piche A. Inflammation-regulating factors in ascites as predictive biomarkers of drug resistance and progression-free survival in serous epithelial ovarian cancers. BMC Cancer 2015; 15: 492
    CrossrefPubMedSearch in Google Scholar
  • 39 Jammal MP, Martins-Filho A, Silveira TP, Murta EF, Nomelini RS. Cytokines and prognostic factors in epithelial ovarian cancer. Clin Med Insights Oncol 2016; 10: 71-76
    PubMedSearch in Google Scholar
  • 40 Ren G, Sun A, Deng C, Zhang J, Wu X, Wei X, Mani S, Dou W, Wang Z. The anti-inflammatory effect and potential mechanism of cardamonin in DSS-induced colitis. Am J Physiol Gastrointest Liver Physiol 2015; 309: G517-G527
    CrossrefPubMedSearch in Google Scholar
  • 41 Israf DA, Khaizurin TA, Syahida A, Lajis NH, Khozirah S. Cardamonin inhibits COX and iNOS expression via inhibition of p65NF-kappaB nuclear translocation and Ikappa-B phosphorylation in RAW 264.7 macrophage cells. Mol Immunol 2007; 44: 673-679
    CrossrefPubMedSearch in Google Scholar
  • 42 Yu H, Jiang W, Du H, Xing Y, Bai G, Zhang Y, Li Y, Jiang H, Zhang Y, Wang J, Wang P, Bai X. Involvement of the Akt/NF-kappaB pathways in the HTNV-mediated increase of IL-6, CCL5, ICAM-1, and VCAM-1 in HUVECs. PLoS One 2014; 9: e93810
    CrossrefPubMedSearch in Google Scholar
  • 43 Wang Q, Huber N, Noel G, Haar L, Shan Y, Pritts TA, Ogle CK. NF-kappaBeta inhibition is ineffective in blocking cytokine-induced IL-8 production but P38 and STAT1 inhibitors are effective. Inflamm Res 2012; 61: 977-985
    CrossrefPubMedSearch in Google Scholar
  • 44 Lin HY, Chang KT, Hung CC, Kuo CH, Hwang SJ, Chen HC, Hung CH, Lin SF. Effects of the mTOR inhibitor rapamycin on monocyte-secreted chemokines. BMC Immunol 2014; 15: 37
    CrossrefPubMedSearch in Google Scholar

Correspondence

Prof. Dr. Daohua Shi
Department of Pharmacy
Fujian Provincial Maternity and Childrenʼs Hospital
18 Daoshan Road, Fuzhou
Fujian, 350001
China   
Phone: + 86 5 91 88 31 20 79   
Fax: + 86 5 91 88 31 20 79   

  • References

  • 1 Chen T, Jansen L, Gondos A, Emrich K, Holleczek B, Katalinic A, Luttmann S, Meyer M, Brenner H. GEKID Cancer Survival Working Group. Survival of ovarian cancer patients in Germany in the early 21st century: a period analysis by age, histology, laterality, and stage. Eur J Cancer Prev 2013; 22: 59-67
    CrossrefPubMedSearch in Google Scholar
  • 2 Edwards HM, Noer MC, Sperling CD, Nguyen-Nielsen M, Lundvall L, Christensen IJ, Hogdall C. Survival of ovarian cancer patients in Denmark: Results from the Danish gynaecological cancer group (DGCG) database, 1995–2012. Acta Oncol 2016; 55 (Suppl. 02) 36-43
    CrossrefPubMedSearch in Google Scholar
  • 3 Xie X, Yang M, Ding Y, Chen J. Microbial infection, inflammation and epithelial ovarian cancer. Oncol Lett 2017; 14: 1911-1919
    CrossrefPubMedSearch in Google Scholar
  • 4 Villar VH, Nguyen TL, Teres S, Bodineau C, Duran RV. Escaping mTOR inhibition for cancer therapy: Tumor suppressor functions of mTOR. Mol Cell Oncol 2017; 4: e1297284
    PubMedSearch in Google Scholar
  • 5 Vangan N, Cao Y, Jia X, Bao W, Wang Y, He Q, Binderiya U, Feng X, Li T, Hao H, Wang Z. mTORC1 mediates peptidoglycan induced inflammatory cytokines expression and NF-κB activation in macrophages. Microb Pathog 2016; 99: 111-118
    CrossrefPubMedSearch in Google Scholar
  • 6 Temiz-Resitoglu M, Kucukkavruk SP, Guden DS, Cecen P, Sari AN, Tunctan B, Gorur A, Tamer-Gumus L, Buharalioglu CK, Malik KU, Sahan-Firat S. Activation of mTOR/IκB-α/NF-κB pathway contributes to LPS-induced hypotension and inflammation in rats. Eur J Pharmacol 2017; 802: 7-19
    CrossrefPubMedSearch in Google Scholar
  • 7 Lee DF, Kuo HP, Chen CT, Hsu JM, Chou CK, Wei Y, Sun HL, Li LY, Ping B, Huang WC, He X, Hung JY, Lai CC, Ding Q, Su JL, Yang JY, Sahin AA, Hortobagyi GN, Tsai FJ, Tsai CH, Hung MC. IKK beta suppression of TSC1 links inflammation and tumor angiogenesis via the mTOR pathway. Cell 2007; 130: 440-455
    CrossrefPubMedSearch in Google Scholar
  • 8 Dufour M, Faes S, Dormond-Meuwly A, Demartines N, Dormond O. PGE2-induced colon cancer growth is mediated by mTORC1. Biochem Biophys Res Comm 2014; 451: 587-591
    CrossrefPubMedSearch in Google Scholar
  • 9 Pan H, Xu LH, Ouyang DY, Wang Y, Zha QB, Hou XF, He XH. The second-generation mTOR kinase inhibitor INK128 exhibits anti-inflammatory activity in lipopolysaccharide-activated RAW 264.7 cells. Inflammation 2014; 37: 756-765
    CrossrefPubMedSearch in Google Scholar
  • 10 Bao W, Wang Y, Fu Y, Jia X, Li J, Vangan N, Bao L, Hao H, Wang Z. mTORC1 regulates flagellin-induced inflammatory response in macrophages. PLoS One 2015; 10: e0125910
    CrossrefPubMedSearch in Google Scholar
  • 11 Sun Q, Liu Q, Zheng Y, Cao X. Rapamycin suppresses TLR4-triggered IL-6 and PGE(2) production of colon cancer cells by inhibiting TLR4 expression and NF-kappaB activation. Mol Immunol 2008; 45: 2929-2936
    CrossrefPubMedSearch in Google Scholar
  • 12 Chandrika G, Natesh K, Ranade D, Chugh A, Shastry P. Suppression of the invasive potential of Glioblastoma cells by mTOR inhibitors involves modulation of NFkappaB and PKC-alpha signaling. Sci Rep 2016; 6: 22455
    CrossrefPubMedSearch in Google Scholar
  • 13 Ekshyyan O, Khandelwal AR, Rong X, Moore-Medlin T, Ma X, Alexander JS, Nathan CO. Rapamycin targets interleukin 6 (IL-6) expression and suppresses endothelial cell invasion stimulated by tumor cells. Am J Transl Res 2016; 8: 4822-4830
    PubMedSearch in Google Scholar
  • 14 Goncalves LM, Valente IM, Rodrigues JA. An overview on cardamonin. J Med Food 2014; 17: 633-640
    CrossrefPubMedSearch in Google Scholar
  • 15 Niu PG, Zhang YX, Shi DH, Liu Y, Chen YY, Deng J. Cardamonin inhibits metastasis of Lewis lung carcinoma cells by decreasing mTOR activity. PLoS One 2015; 10: e0127778
    CrossrefPubMedSearch in Google Scholar
  • 16 Xue ZG, Niu PG, Shi DH, Liu Y, Deng J, Chen YY. Cardamonin inhibits angiogenesis by mTOR downregulation in SKOV3 cells. Planta Med 2016; 82: 70-75
    Thieme ConnectPubMedSearch in Google Scholar
  • 17 Liao Q, Shi DH, Zheng W, Xu XJ, Yu YH. Antiproliferation of cardamonin is involved in mTOR on aortic smooth muscle cells in high fructose-induced insulin resistance rats. Eur J Pharmacol 2010; 641: 179-186
    CrossrefPubMedSearch in Google Scholar
  • 18 Niu P, Zhang Y, Shi D, Chen Y, Deng J. Cardamonin ameliorates insulin resistance induced by high insulin and high glucose through the mTOR and signal pathway. Planta Med 2013; 79: 452-458
    Thieme ConnectPubMedSearch in Google Scholar
  • 19 Lee JH, Jung HS, Giang PM, Jin X, Lee S, Son PT, Lee D, Hong YS, Lee K, Lee JJ. Blockade of nuclear factor-kappaB signaling pathway and anti-inflammatory activity of cardamomin, a chalcone analog from Alpinia conchigera . J Pharmacol Exp Ther 2006; 316: 271-278
    CrossrefPubMedSearch in Google Scholar
  • 20 Chow YL, Lee KH, Vidyadaran S, Lajis NH, Akhtar MN, Israf DA, Syahida A. Cardamonin from Alpinia rafflesiana inhibits inflammatory responses in IFN-gamma/LPS-stimulated BV2 microglia via NF-kappaB signalling pathway. Int Immunopharmacol 2012; 12: 657-665
    CrossrefPubMedSearch in Google Scholar
  • 21 Shan W, Liu J. Inflammation: a hidden path to breaking the spell of ovarian cancer. Cell Cycle 2009; 8: 3107-3111
    CrossrefPubMedSearch in Google Scholar
  • 22 White KL, Schildkraut JM, Palmieri RT, Iversen jr. ES, Berchuck A, Vierkant RA, Rider DN, Charbonneau B, Cicek MS, Sutphen R, Birrer MJ, Pharoah PP, Song H, Tyrer J, Gayther SA, Ramus SJ, Wentzensen N, Yang HP, Garcia-Closas M, Phelan CM, Cunningham JM, Fridley BL, Sellers TA, Goode EL. Ovarian Cancer Association Consortium. Ovarian cancer risk associated with inherited inflammation-related variants. Cancer Res 2012; 72: 1064-1069
    CrossrefPubMedSearch in Google Scholar
  • 23 Cho JS, Kang JH, Um JY, Han IH, Park IH, Lee HM. Lipopolysaccharide induces pro-inflammatory cytokines and MMP production via TLR4 in nasal polyp-derived fibroblast and organ culture. PLoS One 2014; 9: e90683
    CrossrefPubMedSearch in Google Scholar
  • 24 Laird MH, Rhee SH, Perkins DJ, Medvedev AE, Piao W, Fenton MJ, Vogel SN. TLR4/MyD88/PI3K interactions regulate TLR4 signaling. J Leukoc Biol 2009; 85: 966-977
    CrossrefPubMedSearch in Google Scholar
  • 25 Thiem S, Pierce TP, Palmieri M, Putoczki TL, Buchert M, Preaudet A, Farid RO, Love C, Catimel B, Lei Z, Rozen S, Gopalakrishnan V, Schaper F, Hallek M, Boussioutas A, Tan P, Jarnicki A, Ernst M. mTORC1 inhibition restricts inflammation-associated gastrointestinal tumorigenesis in mice. J Clin Invest 2013; 123: 767-781
    PubMedSearch in Google Scholar
  • 26 Deng L, Zhou JF, Sellers RS, Li JF, Nguyen AV, Wang Y, Orlofsky A, Liu Q, Hume DA, Pollard JW, Augenlicht L, Lin EY. A novel mouse model of inflammatory bowel disease links mammalian target of rapamycin-dependent hyperproliferation of colonic epithelium to inflammation-associated tumorigenesis. Am J Pathol 2010; 176: 952-967
    CrossrefPubMedSearch in Google Scholar
  • 27 Li Z, Zhang J, Mulholland M, Zhang W. mTOR activation protects liver from ischemia/reperfusion-induced injury through NF-kappaB pathway. FASEB J 2017; 31: 3018-3026
    CrossrefPubMedSearch in Google Scholar
  • 28 Wang Y, Zhang X, Tang W, Lin Z, Xu L, Dong R, Li Y, Li J, Zhang Z, Li X, Zhao L, Wei JJ, Shao C, Kong B, Liu Z. miR-130a upregulates mTOR pathway by targeting TSC1 and is transactivated by NF-kappaB in high-grade serous ovarian carcinoma. Cell Death Differ 2017; 24: 2089-2100
    CrossrefPubMedSearch in Google Scholar
  • 29 Dan HC, Cooper MJ, Cogswell PC, Duncan JA, Ting JP, Baldwin AS. Akt-dependent regulation of NF-{kappa}B is controlled by mTOR and Raptor in association with IKK. Genes Dev 2008; 22: 1490-1500
    CrossrefPubMedSearch in Google Scholar
  • 30 Sun R, Zhang Y, Ma S, Qi H, Wang M, Duan J, Ma S, Zhu X, Li G, Wang H. Down-regulation of mitogen-activated protein kinases and nuclear factor-kappaB signaling is involved in rapamycin suppression of TLR2-induced inflammatory response in monocytic THP-1 cells. Microbiol Immunol 2015; 59: 614-622
    CrossrefPubMedSearch in Google Scholar
  • 31 Shi D, Zhu Y, Niu P, Zhou J, Chen H. Raptor mediates the antiproliferation of cardamonin by mTORC1 inhibition in SKOV3 cells. Onco Targets Ther 2018; 11: 757-767
    CrossrefPubMedSearch in Google Scholar
  • 32 Morais C, Pat B, Gobe G, Johnson DW, Healy H. Pyrrolidine dithiocarbamate exerts anti-proliferative and pro-apoptotic effects in renal cell carcinoma cell lines. Nephrol Dial Transplant 2006; 21: 3377-3388
    CrossrefPubMedSearch in Google Scholar
  • 33 Wu D, Cheng J, Sun G, Wu S, Li M, Gao Z, Zhai S, Li P, Su D, Wang X. p70S6K promotes IL-6-induced epithelial-mesenchymal transition and metastasis of head and neck squamous cell carcinoma. Oncotarget 2016; 7: 36539-36550
    CrossrefPubMedSearch in Google Scholar
  • 34 Okamoto T, Ozawa Y, Kamoshita M, Osada H, Toda E, Kurihara T, Nagai N, Umezawa K, Tsubota K. The neuroprotective effect of rapamycin as a modulator of the mTOR-NF-kappaB axis during retinal inflammation. PLoS One 2016; 11: e0146517
    CrossrefPubMedSearch in Google Scholar
  • 35 Wendel HG, De Stanchina E, Fridman JS, Malina A, Ray S, Kogan S, Cordon-Cardo C, Pelletier J, Lowe SW. Survival signalling by Akt and eIF4E in oncogenesis and cancer therapy. Nature 2004; 428: 332-337
    PubMedSearch in Google Scholar
  • 36 Park S, Zhao D, Hatanpaa KJ, Mickey BE, Saha D, Boothman DA, Story MD, Wong ET, Burma S, Georgescu MM, Rangnekar VM, Chauncey SS, Habib AA. RIP1 activates PI3K-Akt via a dual mechanism involving NF-kappaB-mediated inhibition of the mTOR-S6K-IRS1 negative feedback loop and down-regulation of PTEN. Cancer Res 2009; 69: 4107-4111
    CrossrefPubMedSearch in Google Scholar
  • 37 Carracedo A, Ma L, Teruya-Feldstein J, Rojo F, Salmena L, Alimonti A, Egia A, Sasaki AT, Thomas G, Kozma SC, Papa A, Nardella C, Cantley LC, Baselga J, Pandolfi PP. Inhibition of mTORC1 leads to MAPK pathway activation through a PI3K-dependent feedback loop in human cancer. J Clin Invest 2008; 118: 3065-3074
    PubMedSearch in Google Scholar
  • 38 Lane D, Matte I, Garde-Granger P, Laplante C, Carignan A, Rancourt C, Piche A. Inflammation-regulating factors in ascites as predictive biomarkers of drug resistance and progression-free survival in serous epithelial ovarian cancers. BMC Cancer 2015; 15: 492
    CrossrefPubMedSearch in Google Scholar
  • 39 Jammal MP, Martins-Filho A, Silveira TP, Murta EF, Nomelini RS. Cytokines and prognostic factors in epithelial ovarian cancer. Clin Med Insights Oncol 2016; 10: 71-76
    PubMedSearch in Google Scholar
  • 40 Ren G, Sun A, Deng C, Zhang J, Wu X, Wei X, Mani S, Dou W, Wang Z. The anti-inflammatory effect and potential mechanism of cardamonin in DSS-induced colitis. Am J Physiol Gastrointest Liver Physiol 2015; 309: G517-G527
    CrossrefPubMedSearch in Google Scholar
  • 41 Israf DA, Khaizurin TA, Syahida A, Lajis NH, Khozirah S. Cardamonin inhibits COX and iNOS expression via inhibition of p65NF-kappaB nuclear translocation and Ikappa-B phosphorylation in RAW 264.7 macrophage cells. Mol Immunol 2007; 44: 673-679
    CrossrefPubMedSearch in Google Scholar
  • 42 Yu H, Jiang W, Du H, Xing Y, Bai G, Zhang Y, Li Y, Jiang H, Zhang Y, Wang J, Wang P, Bai X. Involvement of the Akt/NF-kappaB pathways in the HTNV-mediated increase of IL-6, CCL5, ICAM-1, and VCAM-1 in HUVECs. PLoS One 2014; 9: e93810
    CrossrefPubMedSearch in Google Scholar
  • 43 Wang Q, Huber N, Noel G, Haar L, Shan Y, Pritts TA, Ogle CK. NF-kappaBeta inhibition is ineffective in blocking cytokine-induced IL-8 production but P38 and STAT1 inhibitors are effective. Inflamm Res 2012; 61: 977-985
    CrossrefPubMedSearch in Google Scholar
  • 44 Lin HY, Chang KT, Hung CC, Kuo CH, Hwang SJ, Chen HC, Hung CH, Lin SF. Effects of the mTOR inhibitor rapamycin on monocyte-secreted chemokines. BMC Immunol 2014; 15: 37
    CrossrefPubMedSearch in Google Scholar

   Permissions and Reprints
Zoom Image
Fig. 1 The chemical structure of cardamonin.
Zoom Image
Fig. 2 Cardamonin effects on the cell viability of normal and LPS-pretreated SKOV3 cells. Cells were pretreated with or without LPS (1 µg/mL) for 30 min, and then treated with cardamonin (1, 3, 10, 30, and 100 µM) and rapamycin (0.1 µM) for 48 h, respectively. Cell viability was then measured by the MTT assay (n = 5). Data are presented as the mean ± SEM of five experiments. *P < 0.05, **p < 0.01 compared with the control group; #p < 0.05, ##p < 0.01 compared with the LPS-pretreated group.
Zoom Image
Fig. 3 Cardamonin inhibited the expression of TNF-α and IL-6. SKOV3 cells were pretreated with or without LPS (1 µg/mL) for 30 min, and then treated with cardamonin (3 or 30 µM) and rapamycin (0.1 µM) for 24 h, respectively. Secreted concentrations of TNF-α, IL-6, and IL-8 in the media were determined by ELISA (n = 5). A Concentration of TNF-α. B Concentration of IL-6. C Concentration of IL-8. Data are presented by the mean ± SEM of five experiments. **P < 0.01 compared with the control group; ##p < 0.01 compared with the LPS-pretreated group.
Zoom Image
Fig. 4 Cardamonin decreased the activation of NF-κB p65. SKOV3 cells were pretreated with or without LPS (1 µg/mL) for 30 min, and then treated with cardamonin (3 or 30 µM) and rapamycin (0.1 µM) for 24 h, respectively. Total protein and nuclear proteins were extracted for Western blot analysis (n = 3). A The protein bands of total NF-κB p65, nuclear NF-κB p65, and histone H2A. B The relative density ratio of nuclear NF-κB p65 was normalized to histone H2A. Data are presented as the mean ± SEM of three experiments. **P < 0.01 compared with the control group; ##p < 0.01 compared with the LPS-pretreated group.
Zoom Image
Fig. 5 Cardamonin decreased the activation of mTOR and S6K1. SKOV3 cells were pretreated with or without LPS (1 µg/mL) for 30 min, and then treated with cardamonin (3 or 30 µM) and rapamycin (0.1 µM) for 24 h, respectively. Total protein was extracted for Western blot analysis (n = 3). A The protein bands of mTOR, p-mTOR, S6K1, p-S6K1, Akt, p-Akt, and actin. B The relative density ratios of p-mTOR, p-S6K1, and p-Akt protein were normalized to actin. Data are presented as the mean ± SEM of three experiments. *P < 0.05, **p < 0.01 compared with the control group; #p < 0.05, ##p < 0.01 compared with the LPS-pretreated group.
Zoom Image
Fig. 6 Effects of cardamonin on the expression of TNF-α, IL-6, and IL-8 in mTOR and NF-κB inhibitor pretreated cells. Cells were cultured with rapamycin (0.1 µM) and PDTC (100 µM) for 1 h, respectively, and then treated with LPS (1 µg/mL) for 30 min; lastly, cells were treated with cardamonin (30 µM) for 24 h. Secreted concentrations of TNF-α, IL-6, and IL-8 in the media were determined by ELISA (n = 5). A Concentration of TNF-α. B Concentration of IL-6. C Concentration of IL-8. Data are presented as the mean ± SEM of five experiments. **P < 0.01 compared with the control group; ##p < 0.01 compared with the LPS-pretreated group; &&p < 0.05 compared with the LPS + cardamonin group.
Zoom Image
Fig. 7 Effects of cardamonin on the activation of NF-κB p65 in mTOR and NF-κB inhibitor pretreated cells. Cells were cultured with rapamycin (0.1 µM) and PDTC (100 µM) for 1 h, respectively, and then treated with LPS (1 µg/mL) for 30 min; lastly, cells were treated with cardamonin (30 µM) for 24 h. Total protein and nuclear proteins were extracted for Western blot analysis (n = 3). A The protein bands of total NF-κB p65, nuclear NF-κB p65, and histone H2A. B The relative density ratio of nuclear NF-κB p65 was normalized to histone H2A. Data are presented as the mean ± SEM of three experiments. **P < 0.01 compared with the control group; ##p < 0.01 compared with the LPS-pretreated group; &&p < 0.05 compared with the LPS + cardamonin group.
Zoom Image
Fig. 8 Effects of cardamonin on the activation of mTOR and S6K1 in mTOR and NF-κB inhibitor pretreated cells. Cells were cultured with rapamycin (0.1 µM) and PDTC (100 µM) for 1 h, respectively, and then treated with LPS (1 µg/mL) for 30 min; lastly, cells were treated with cardamonin (30 µM) for 24 h. Total protein was extracted for Western blot analysis (n = 3). A The protein bands of mTOR, p-mTOR, S6K1, p-S6K1, Akt, p-Akt, and actin. B The relative density ratios of p-mTOR, p-S6K1, and p-Akt proteins were normalized to actin. Data are presented as the mean ± SEM of three experiments. **P < 0.01 compared with the control group; #p < 0.05, ##p < 0.01 compared with the LPS-pretreated group; &&p < 0.05 compared with the LPS + cardamonin group.