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Ponicidin inhibits pro-inflammatory cytokine TNF-α-induced epithelial–mesenchymal transition and metastasis of colorectal cancer cells via suppressing the AKT/GSK-3β/Snail pathway

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Abstract

Ponicidin (PON), a natural diterpenoid compound, has been shown to exhibit potent anticancer activities in a wide variety of cancers, including colorectal cancer (CRC). Nevertheless, the precise mechanisms underlying the anti-metastasis effect of PON have not yet been completely defined. The present study was designed to uncover the inhibitory effect of PON on epithelial–mesenchymal transition (EMT), migration and invasion of HCT116 cells induced by pro-inflammatory cytokine tumor necrosis factor-α (TNF-α) in vitro, and liver metastasis in vivo. Briefly, cell proliferation was assessed by Cell Counting Kit-8 assay, followed by wound healing and transwell assays to evaluate cell migration and invasion. The EMT-related molecular markers were determined through quantitative real-time polymerase chain reaction (qPCR), immunofluorescence (IF), western blot (WB), and immunohistochemistry (IHC). Additionally, WB was used to assess the expression of AKT, phosphorylated AKT (p-AKT), GSK-3β, and phosphorylated GSK-3β (p-GSK-3β). As a result, PON could effectively suppress EMT, migration, and invasion in HCT116 cells in vitro, and liver metastasis of HCT116 cells in vivo. Additionally, PON administration also dramatically altered the expression of EMT-associated markers such as E-cadherin, N-cadherin, and Vimentin, and suppressed the expression of p-AKT, p-GSK-3β and transcription factor, Snail in a dose-dependent manner. Moreover, the incidence of liver metastasis in the control group was 100% and although the incidence of liver metastasis did not decrease, the number of metastatic nodules in the livers of each PON dose group decreased by (34 ± 4.2)%, (64 ± 3.6)%, and (76 ± 5.3)%, respectively, compared to the control group. Collectively, these findings indicated that targeting the AKT/GSK-3β/Snail pathway by PON might be a promising treatment for TNF-α-induced EMT and metastasis of CRC.

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References

  • Balkwill F (2009) Tumour necrosis factor and cancer. Nat Rev Cancer 9:361–371

    Article  CAS  PubMed  Google Scholar 

  • Bertazza L, Mocellin S (2010) The dual role of tumor necrosis factor (TNF) in cancer biology. Curr Med Chem 17:3337–3352

    Article  CAS  PubMed  Google Scholar 

  • Broster SA, Kyprianou N (2015) Epithelial–mesenchymal transition in prostatic disease. Future Oncol 11:3197–3206

    Article  CAS  PubMed  Google Scholar 

  • Cao H, Xu E, Liu H, Wan L, Lai M (2015) Epithelial–mesenchymal transition in colorectal cancer metastasis: a system review. Pathol Res Pract 211:557–569

    Article  CAS  PubMed  Google Scholar 

  • Cheng Z, Guo Y, Yang Y, Kan J, Dai S, Helian M, Li B, Xu J, Liu C (2016) Nitidine chloride suppresses epithelial-to-mesenchymal transition in osteosarcoma cell migration and invasion through Akt/GSK-3β/Snail signaling pathway. Oncol Rep 36:1023–1029

    Article  CAS  PubMed  Google Scholar 

  • Du J, Chen C, Sun Y, Zheng L, Wang W (2015) Ponicidin suppresses HT29 cell growth via the induction of G1 cell cycle arrest and apoptosis. Mol Med Rep 12:5816–5820

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Fan F, Samuel S, Evans KW, Lu J, Xia L, Zhou Y, Sceusi E, Tozzi F, Ye XC, Mani SA, Ellis LM (2012) Overexpression of snail induces epithelial–mesenchymal transition and a cancer stem cell-like phenotype in human colorectal cancer cells. Cancer Med 1:5–16

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ginnebaugh KR, Ahmad A, Sarkar FH (2014) The therapeutic potential of targeting the epithelial–mesenchymal transition in cancer. Expert Opin Ther Targets 18:731–745

    Article  CAS  PubMed  Google Scholar 

  • Guo H, Luo H, Yuan H, Xia Y, Shu P, Huang X, Lu Y, Liu X, Keller ET, Sun D, Deng J, Zhang J (2017a) Litchi seed extracts diminish prostate cancer progression via induction of apoptosis and attenuation of EMT through Akt/GSK-3β signaling. Sci Rep 7:41656

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Guo S, Cui X, Jiang M, Bai L, Tian X, Guo T, Liu Q, Zhang L, Ho CT, Bai N (2017b) Simultaneous characterization and quantification of 17 main compounds in Rabdosia rubescens by high performance liquid chromatography. J Food Drug Anal 25:417–424

    Article  CAS  PubMed  Google Scholar 

  • Hayashi K, Hayashi T, Sun HD, Takeda Y (2002) Contribution of a combination of ponicidin and acyclovir/ganciclovir to the antitumor efficacy of the herpes simplex virus thymidine kinase gene therapy system. Hum Gene Ther 13:415–423

    Article  CAS  PubMed  Google Scholar 

  • Ho MY, Tang SJ, Chuang MJ, Cha TL, Li JY, Sun GH, Sun KH (2012) TNF-α induces epithelial–mesenchymal transition of renal cell carcinoma cells via a GSK3β-dependent mechanism. Mol Cancer Res 10:1109–1119

    Article  CAS  PubMed  Google Scholar 

  • Hsieh TC, Wijeratne EK, Liang JY, Gunatilaka AL, Wu JM (2005) Differential control of growth, cell cycle progression, and expression of NF-kappaB in human breast cancer cells MCF-7, MCF-10A, and MDA-MB-231 by ponicidin and oridonin, diterpenoids from the Chinese herb Rabdosia rubescens. Biochem Biophys Res Commun 337:224–231

    Article  CAS  PubMed  Google Scholar 

  • Jin H, Yu Y, Zhang T, Zhou X, Zhou J, Jia L, Wu Y, Zhou BP, Feng Y (2010) Snail is critical for tumor growth and metastasis of ovarian carcinoma. Int J Cancer 126:2102–2111

    CAS  PubMed  Google Scholar 

  • Kim YH, Kim G, Kwon CI, Kim JW, Park PW, Hahm KB, Upur H (2014) TWIST1 and SNAI1 as markers of poor prognosis in human colorectal cancer are associated with the expression of ALDH1 and TGF-β1. Oncol Rep 31:1380–1388

    Article  CAS  PubMed  Google Scholar 

  • Kwon CH, Park HJ, Choi JH, Lee JR, Kim HK, Jo HJ, Kim HS, Oh N, Song GA, Park DY (2015) Snail and serpinA1 promote tumor progression and predict prognosis in colorectal cancer. Oncotarget 6:20312–20326

    PubMed  PubMed Central  Google Scholar 

  • Lee JG, Ko MK, Kay EP (2012) Endothelial mesenchymal transformation mediated by IL-1β-induced FGF-2 in corneal endothelial cells. Exp Eye Res 95:35–39

    Article  CAS  PubMed  Google Scholar 

  • Liu JJ, Zhang Y, Guang WB, Yang HZ, Lin DJ, Xiao RZ (2008) Ponicidin inhibits monocytic leukemia cell growth by induction of apoptosis. Int J Mol Sci 9:2265–2277

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Liu ZC, Wang HS, Zhang G, Liu H, Chen XH, Zhang F, Chen DY, Cai SH, Du J (2014) AKT/GSK-3β regulates stability and transcription of snail which is crucial for bFGF-induced epithelial–mesenchymal transition of prostate cancer cells. Biochim Biophys Acta 1840:3096–3105

    Article  CAS  PubMed  Google Scholar 

  • Liu YF, Lu YM, Qu GQ, Liu Y, Chen WX, Liao XH, Kong WM (2015) Ponicidin induces apoptosis via JAK2 and STAT3 signaling pathways in gastric carcinoma. Int J Mol Sci 16:1576–1589

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Liu H, Wang N, Zhang Z, Wang H, Du J, Tang J (2017) Effects of tumor necrosis factor-α on morphology and mechanical properties of HCT116 human colon cancer cells investigated by atomic force microscopy. Scanning 2017:2027079

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Lu H, Chen B, Hong W, Liang Y, Bai Y (2016) Transforming growth factor-β1 stimulates hedgehog signaling to promote epithelial–mesenchymal transition after kidney injury. FEBS J 283:3771–3790

    Article  CAS  PubMed  Google Scholar 

  • Miao JW, Liu LJ, Huang J (2014) Interleukin-6-induced epithelial–mesenchymal transition through signal transducer and activator of transcription 3 in human cervical carcinoma. Int J Oncol 45:165–176

    Article  CAS  PubMed  Google Scholar 

  • Nieto MA (2002) The snail superfamily of zinc-finger transcription factors. Nat Rev Mol Cell Biol 3:155–166

    Article  CAS  PubMed  Google Scholar 

  • Nishioka R, Itoh S, Gui T, Gai Z, Oikawa K, Kawai M, Tani M, Yamaue H, Muragaki Y (2010) SNAIL induces epithelial-to-mesenchymal transition in a human pancreatic cancer cell line (BxPC3) and promotes distant metastasis and invasiveness in vivo. Exp Mol Pathol 89:149–157

    Article  CAS  PubMed  Google Scholar 

  • Peng Z, Wang CX, Fang EH, Wang GB, Tong Q (2014) Role of epithelial–mesenchymal transition in gastric cancer initiation and progression. World J Gastroenterol 20:5403–5410

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Roy HK, Smyrk TC, Koetsier J, Victor TA, Wali RK (2005) The transcriptional repressor SNAIL is overexpressed in human colon cancer. Dig Dis Sci 50:42–46

    Article  CAS  PubMed  Google Scholar 

  • Shin NR, Jeong EH, Choi CI, Moon HJ, Kwon CH, Chu IS, Kim GH, Jeon TY, Kim DH, Lee JH, Park DY (2012) Overexpression of Snail is associated with lymph node metastasis and poor prognosis in patients with gastric cancer. BMC Cancer 12:521–535

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Siegel RL, Miller KD, Fedewa SA, Ahnen DJ, Meester RGS, Barzi A, Jemal A (2017) Colorectal cancer statistics, 2017. CA Cancer J Clin 67:177–193

    Article  PubMed  Google Scholar 

  • Techasen A, Namwat N, Loilome W, Bungkanjana P, Khuntikeo N, Puapairoj A, Jearanaikoon P, Saya H, Yongvanit P (2012) Tumor necrosis factor-α (TNF-α) stimulates the epithelial–mesenchymal transition regulator Snail in cholangiocarcinoma. Med Oncol 29:3083–3091

    Article  CAS  PubMed  Google Scholar 

  • Techasen A, Namwat N, Loilome W, Duangkumpha K, Puapairoj A, Saya H, Yongvanit P (2014) Tumor necrosis factor-α modulates epithelial mesenchymal transition mediators ZEB2 and S100A4 to promote cholangiocarcinoma progression. J Hepatobiliary Pancreat Sci 21:703–711

    Article  PubMed  Google Scholar 

  • Valderrama-Treviño AI, Barrera-Mera B, Ceballos-Villalva JC, Montalvo-Javé EE (2017) Hepatic metastasis from colorectal cancer. Euroasian J Hepatogastroenterol 7:166–175

    Article  PubMed  PubMed Central  Google Scholar 

  • Wang Y, Zhou BP (2011) Epithelial–mesenchymal transition in breast cancer progression and metastasis. Chin J Cancer 30:603–611

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wang H, Fang R, Wang XF, Zhang F, Chen DY, Zhou B, Wang HS, Cai SH, Du J (2013a) Stabilization of Snail through AKT/GSK-3β signaling pathway is required for TNF-α-induced epithelial–mesenchymal transition in prostate cancer PC3 cells. Eur J Pharmacol 714:48–55

    Article  CAS  PubMed  Google Scholar 

  • Wang H, Wang HS, Zhou BH, Li CL, Zhang F, Wang XF, Zhang G, Bu XZ, Cai SH, Du J (2013b) Epithelial–mesenchymal transition (EMT) induced by TNF-α requires AKT/GSK-3β-mediated stabilization of Snail in colorectal cancer. PLoS One 8(2):e56664

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wang Y, Shi J, Chai K, Ying X, Zhou BP (2013c) The role of Snail in EMT and tumorigenesis. Curr Cancer Drug Targets 13:963–972

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Xie W, Jin Y, Hou L, Ma Y, Xu H, Zhang K, Zhang L, Du Y (2017) A practical strategy for the characterization of ponicidin metabolites in vivo and in vitro by UHPLC-Q-TOF-MS based on nontargeted SWATH data acquisition. J Pharm Biomed Anal 145:865–878

    Article  CAS  PubMed  Google Scholar 

  • Yin T, Wang C, Liu T, Zhao G, Zha Y, Yang M (2007) Expression of snail in pancreatic cancer promotes metastasis and chemoresistance. J Surg Res 141:196–203

    Article  CAS  PubMed  Google Scholar 

  • Zhang JF, Liu PQ, Chen GH, Lu MQ, Cai CJ, Yang Y, Li H (2007) Ponicidin inhibits cell growth on hepatocellular carcinoma cells by induction of apoptosis. Dig Liver Dis 39:160–166

    Article  CAS  PubMed  Google Scholar 

  • Zhou ZJ, Dai Z, Zhou SL, Hu ZQ, Chen Q, Zhao YM, Shi YH, Gao Q, Wu WZ, Qiu SJ, Zhou J, Fan J (2014) HNRNPAB induces epithelial–mesenchymal transition and promotes metastasis of hepatocellular carcinoma by transcriptionally activating SNAIL. Cancer Res 74:2750–2762

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

The authors are grateful to Jiapeng Zhu and Cunsi Shen for the financial support. The specific support funds are National key R & D Plan for Precision Medicine Research (2016YFC0905900) (Jiapeng Zhu), Research Funds for Special Professors of Jiangsu Province of China (No. 013038009001) (Jiapeng Zhu) and Natural Science Foundation of Jiangsu Province of China (No. BK20151004) (Cunsi Shen).

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Authors and Affiliations

  1. School of Medicine and Life Sciences, Nanjing University of Chinese Medicine, Nanjing, 210023, China

    Zhengguang Zhang, Jingjing Xu, Bing Liu, Feiyan Chen, Jiao Li, Yuchen Liu & Jiapeng Zhu

  2. Jiangsu Key Laboratory of Pediatric Respiratory Disease, Institute of Pediatics, Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing, 210023, China

    Zhengguang Zhang & Cunsi Shen

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  1. Zhengguang Zhang
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Zhang, Z., Xu, J., Liu, B. et al. Ponicidin inhibits pro-inflammatory cytokine TNF-α-induced epithelial–mesenchymal transition and metastasis of colorectal cancer cells via suppressing the AKT/GSK-3β/Snail pathway. Inflammopharmacol 27, 627–638 (2019). https://doi.org/10.1007/s10787-018-0534-5

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