Triptolide inhibits viability and induces apoptosis in liver cancer cells through activation of the tumor suppressor gene p53

Sun,Yan-Yan; Xiao,Lei; Wang,Dong; Ji,Yan-Chao; Yang,Yu-Peng; Ma,Rong; Chen,Xi-Hai

doi:10.3892/ijo.2017.3850

Triptolide inhibits viability and induces apoptosis in liver cancer cells through activation of the tumor suppressor gene p53

Authors:
- Yan-Yan Sun
- Lei Xiao
- Dong Wang
- Yan-Chao Ji
- Yu-Peng Yang
- Rong Ma
- Xi-Hai Chen
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Affiliations: Department of General Surgery, The Fourth Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150001, P.R. China, Department of Gynecological Oncology, The Third Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150040, P.R. China
Published online on: January 16, 2017 https://doi.org/10.3892/ijo.2017.3850
Pages: 847-852

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Abstract

The present study investigated the effect of triptolide on viability and apoptosis along with underlying mechanism in liver cancer cells. CCK-8 assay showed that triptolide treatment for 48 h significantly reduced the viability of HepG2 and QSG7701 cells at 50 µM concentration. Annexin V-FITC and propidium iodide staining showed that triptolide treatment of HepG2 cells at 50 µM concentrations induced apoptosis in 56.45% cells compared to only 2.36% cells in the control cultures. Western blot assay showed that treatment of HepG2 cells with 50 µM concentration of triptolide significantly induced phosphorylation of p53 in a 2 h-treatment. Phosphorylation of histone H2A.X indicator of DNA damage was induced by triptolide treatment after 12 h in HepG2 cells. The level of nuclear p53 in a 6 h-treatment with 0, 10, 20, 30, 40 and 50 µM concentration of triptolide was found to be 15.3, 19.6, 28.5, 43.7, 63.8 and 91.5%, respectively. Treatment of HepG2 cells with triptolide at 50 µM concentration caused a significant increase in the binding potential of p53 to DNA. Triptolide treatment of HepG2 cells caused a significant increase in the expression of p21, Bax and DR5 genes in HepG2 cells. It also increased the expression of miR-34b and miR-34c in HepG2 cells markedly. Treatment of HepG2 cells with p53 inhibitor, pifithrin-α prior to incubation with triptolide significantly prevented induction of cell apoptosis. Suppression of p53 expression by siRNA inhibited the expression of p53 as well as its target genes along with the prevention of apoptosis induction. In conclusion, triptolide inhibits viability and induces apoptosis in liver cancer cells through activation of the tumor suppressor gene p53. Thus, triptolide can be used for the treatment of liver cancer.

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1	Zhen QS, Ye X and Wei ZJ: Recent progress in research on Tripterygium: A male antifertility plant. Contraception. 51:121–129. 1995. View Article : Google Scholar : PubMed/NCBI
2	Tengchaisri T, Chawengkirttikul R, Rachaphaew N, Reutrakul V, Sangsuwan R and Sirisinha S: Antitumor activity of triptolide against cholangiocarcinoma growth in vitro and in hamsters. Cancer Lett. 133:169–175. 1998. View Article : Google Scholar
3	Hachida M, Lu H, Zhang X, Saito S, Furutani Y, Matsuoka R, Hoshi H and Koyanagi H: Inhibitory effect of triptolide on platelet derived growth factor-A and coronary arteriosclerosis after heart transplantation. Transplant Proc. 31:2719–2723. 1999. View Article : Google Scholar : PubMed/NCBI
4	Carter BZ, Mak DH, Schober WD, McQueen T, Harris D, Estrov Z, Evans RL and Andreeff M: Triptolide induces caspase-dependent cell death mediated via the mitochondrial pathway in leukemic cells. Blood. 108:630–637. 2006. View Article : Google Scholar : PubMed/NCBI
5	Cohen JJ: Apoptosis. Immunol Today. 14:126–130. 1993. View Article : Google Scholar : PubMed/NCBI
6	White E: Life, death, and the pursuit of apoptosis. Genes Dev. 10:1–15. 1996. View Article : Google Scholar : PubMed/NCBI
7	Williams GT and Smith CA: Molecular regulation of apoptosis: Genetic controls on cell death. Cell. 74:777–779. 1993. View Article : Google Scholar : PubMed/NCBI
8	Wyllie AH, Kerr JF and Currie AR: Cell death: The significance of apoptosis. Int Rev Cytol. 68:251–306. 1980. View Article : Google Scholar : PubMed/NCBI
9	Ma S, Chan KW and Guan XY: In search of liver cancer stem cells. Stem Cell Rev. 4:179–192. 2008. View Article : Google Scholar : PubMed/NCBI
10	Tomuleasa C, Soritau O, Rus-Ciuca D, Pop T, Todea D, Mosteanu O, Pintea B, Foris V, Susman S, Kacso G, et al: Isolation and characterization of hepatic cancer cells with stem-like properties from hepatocellular carcinoma. J Gastrointestin Liver Dis. 19:61–67. 2010.PubMed/NCBI
11	Ricci-Vitiani L, Lombardi DG, Pilozzi E, Biffoni M, Todaro M, Peschle C and De Maria R: Identification and expansion of human colon-cancer-initiating cells. Nature. 445:111–115. 2007. View Article : Google Scholar
12	Lee TK, Castilho A, Ma S and Ng IO: Liver cancer stem cells: Implications for a new therapeutic target. Liver Int. 29:955–965. 2009. View Article : Google Scholar : PubMed/NCBI
13	Vogelstein B, Lane D and Levine AJ: Surfing the p53 network. Nature. 408:307–310. 2000. View Article : Google Scholar : PubMed/NCBI
14	Ma L, Wagner J, Rice JJ, Hu W, Levine AJ and Stolovitzky GA: A plausible model for the digital response of p53 to DNA damage. Proc Natl Acad Sci USA. 102:14266–14271. 2005. View Article : Google Scholar : PubMed/NCBI
15	Harris SL and Levine AJ: The p53 pathway: Positive and negative feedback loops. Oncogene. 24:2899–2908. 2005. View Article : Google Scholar : PubMed/NCBI
16	Levine AJ: p53, the cellular gatekeeper for growth and division. Cell. 88:323–331. 1997. View Article : Google Scholar : PubMed/NCBI
17	Bean LJ and Stark GR: Phosphorylation of serines 15 and 37 is necessary for efficient accumulation of p53 following irradiation with UV. Oncogene. 20:1076–1084. 2001. View Article : Google Scholar : PubMed/NCBI
18	She QB, Chen N and Dong Z: ERKs and p38 kinase phosphorylate p53 protein at serine 15 in response to UV radiation. J Biol Chem. 275:20444–20449. 2000. View Article : Google Scholar : PubMed/NCBI
19	Vazquez A, Bond EE, Levine AJ and Bond GL: The genetics of the p53 pathway, apoptosis and cancer therapy. Nat Rev Drug Discov. 7:979–987. 2008. View Article : Google Scholar : PubMed/NCBI
20	Yin Y, Terauchi Y, Solomon GG, Aizawa S, Rangarajan PN, Yazaki Y, Kadowaki T and Barrett JC: Involvement of p85 in p53-dependent apoptotic response to oxidative stress. Nature. 391:707–710. 1998. View Article : Google Scholar : PubMed/NCBI
21	Lowe SW, Ruley HE, Jacks T and Housman DE: p53-dependent apoptosis modulates the cytotoxicity of anticancer agents. Cell. 74:957–967. 1993. View Article : Google Scholar : PubMed/NCBI
22	Brugarolas J, Chandrasekaran C, Gordon JI, Beach D, Jacks T and Hannon GJ: Radiation-induced cell cycle arrest compromised by p21 deficiency. Nature. 377:552–557. 1995. View Article : Google Scholar : PubMed/NCBI
23	Miyashita T and Reed JC: Tumor suppressor p53 is a direct transcriptional activator of the human bax gene. Cell. 80:293–299. 1995. View Article : Google Scholar : PubMed/NCBI
24	Oda E, Ohki R, Murasawa H, Nemoto J, Shibue T, Yamashita T, Tokino T, Taniguchi T and Tanaka N: Noxa, a BH3-only member of the Bcl-2 family and candidate mediator of p53-induced apoptosis. Science. 288:1053–1058. 2000. View Article : Google Scholar : PubMed/NCBI
25	Yu J, Zhang L, Hwang PM, Kinzler KW and Vogelstein B: PUMA induces the rapid apoptosis of colorectal cancer cells. Mol Cell. 7:673–682. 2001. View Article : Google Scholar : PubMed/NCBI
26	Shieh SY, Ikeda M, Taya Y and Prives C: DNA damage-induced phosphorylation of p53 alleviates inhibition by MDM2. Cell. 91:325–334. 1997. View Article : Google Scholar : PubMed/NCBI
27	Prives C: Signaling to p53: Breaking the MDM2-p53 circuit. Cell. 95:5–8. 1998. View Article : Google Scholar : PubMed/NCBI
28	Brew CT, Aronchik I, Hsu JC, Sheen JH, Dickson RB, Bjeldanes LF and Firestone GL: Indole-3-carbinol activates the ATM signaling pathway independent of DNA damage to stabilize p53 and induce G1 arrest of human mammary epithelial cells. Int J Cancer. 118:857–868. 2006. View Article : Google Scholar
29	Loehberg CR, Thompson T, Kastan MB, Maclean KH, Edwards DG, Kittrell FS, Medina D, Conneely OM and O'Malley BW: Ataxia telangiectasia-mutated and p53 are potential mediators of chloroquine-induced resistance to mammary carcinogenesis. Cancer Res. 67:12026–12033. 2007. View Article : Google Scholar : PubMed/NCBI
30	Jiang C, Hu H, Malewicz B, Wang Z and Lü J: Selenite-induced p53 Ser-15 phosphorylation and caspase-mediated apoptosis in LNCaP human prostate cancer cells. Mol Cancer Ther. 3:877–884. 2004.PubMed/NCBI
31	Zhao R, Xiang N, Domann FE and Zhong W: Expression of p53 enhances selenite-induced superoxide production and apoptosis in human prostate cancer cells. Cancer Res. 66:2296–2304. 2006. View Article : Google Scholar : PubMed/NCBI
32	Chang TC, Wentzel EA, Kent OA, Ramachandran K, Mullendore M, Lee KH, Feldmann G, Yamakuchi M, Ferlito M, Lowenstein CJ, et al: Transactivation of miR-34a by p53 broadly influences gene expression and promotes apoptosis. Mol Cell. 26:745–752. 2007. View Article : Google Scholar : PubMed/NCBI