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

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.

Key words

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

• 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
• 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
• 3 Xie X, Yang M, Ding Y, Chen J. Microbial infection, inflammation and epithelial ovarian cancer. Oncol Lett 2017; 14: 1911-1919
• 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
• 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
• 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
• 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
• 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
• 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
• 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
• 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
• 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
• 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
• 14 Goncalves LM, Valente IM, Rodrigues JA. An overview on cardamonin. J Med Food 2014; 17: 633-640
• 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
• 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
• 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
• 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
• 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
• 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
• 21 Shan W, Liu J. Inflammation: a hidden path to breaking the spell of ovarian cancer. Cell Cycle 2009; 8: 3107-3111
• 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
• 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
• 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
• 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
• 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
• 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
• 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
• 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
• 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
• 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
• 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
• 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
• 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
• 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
• 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
• 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
• 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
• 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
• 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
• 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
• 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
• 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
• 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