As2O3联合γ分泌酶抑制剂对HepG2细胞凋亡的诱导作用及其机制

doi:10.13481/j.1671-587x.20150307

摘要/Abstract

摘要：

目的:探讨As₂O₃通过Notch信号通路对HepG₂细胞凋亡的影响,阐明其作用机制。方法:体外培养的HepG₂细胞分为空白组、As₂O₃组、MW167组和联合组;采用不同浓度As₂O₃和MW167处理HepG₂细胞24、48和72 h,MTT法检测细胞增殖抑制率。根据MTT结果选取无细胞毒剂量As₂O₃和MW167(As₂O₃ 0.25 mg·L^-1、MW167 10 μmol·L^-1)分组干预HepG₂细胞48 h;流式细胞术检测细胞凋亡率;RT-PCR和Western blotting法检测Notch-1、Bcl-2和Caspase-3 mRNA和蛋白表达水平。结果:不同浓度As₂O₃和MW167处理HepG₂细胞均可抑制细胞增殖,呈剂量效应关系;无细胞毒剂量As₂O₃和MW167处理不同分组HepG₂细胞48 h后,与空白组比较,As₂O₃组、MW167组和联合组细胞凋亡率升高(P<0.05),其中以联合组升高最为明显;与空白组比较,As₂O₃组、MW167组和联合组细胞中Bcl-2 、Notch-1 mRNA和蛋白表达水平降低(P<0.05),其中以联合组降低最为明显;与空白组比较,As₂O₃组、MW167组和联合组细胞中Caspase-3 mRNA和蛋白表达水平升高(P<0.05),其中以联合组升高最为明显。结论:As₂O₃可诱导肝癌HepG₂细胞凋亡,其作用机制可能与通过Notch信号通路下调细胞中Bcl-2 、Notch-1基因和蛋白表达水平以及上调细胞中Caspase-3基因和蛋白表达水平有关联。

关键词: HepG₂细胞, &gamma, 分泌酶抑制剂, 三氧化二砷, 细胞凋亡

Abstract:

Objective To discuss the effect of As₂O₃ on the apoptosis of HepG₂ cells via Notch signal pathway, and to clarify its mechanism. Methods The HepG₂ cells cultured in vitro were divided into blank group, As₂O₃ group, MW167 group and combination group.The HepG₂ cells were treated with As₂O₃ and MW167 at different concentrations for 24, 48 and 72 h, and the inhibitory rates of proliferations of the HepG₂ cells were detected by MTT method.The HepG₂ cells in various groups were intervened by MW167 and As₂O₃ at non-cytotoxic doses (As₂O₃ 0.25 mg·L^-1, MW167 10 μmol·L^-1) based on The Results of MTT for 48 h, then the apoptotic rate was tested by flow cytometry;RT-PCR and Western blotting Methods were employed to detect the expressions of Notch-1, Bcl-2 and Caspase-3 mRNA and protein. Results The HepG₂ cells treated with both As₂O₃ and MW167 at different concentrations could inhibit the cell proliferation and showed a dose-effect relationship.For the HepG₂ cells in different groups treated with MW167 group and As₂O₃ group at non-cytotoxic doses for 48 h, compared with blank group, the apoptotic rates in As₂O₃ group, MW167 group and combination group were increased(P<0.05), especially in combination group; the expression levels of Bcl-2, Notch-1 mRNA and protein in As₂O₃ group, MW167 group and combination group were lower than those in blank group(P<0.05), especially in combination group;compared with blank group, the expressions levels of Caspase-3 mRNA and protein in As₂O₃ group, MW167 group and combination group were increased(P<0.05), especially in combination group. Conclusion As₂O₃ can induce the apoptosis of hepatocelular carcinoma HepG₂ cells and the mechanism may be related to the down-regulation of Bcl-2 and Notch-1 and up-regulation of Caspase-3 mRNA and protein through Notch signal pathway.

Key words: HepG₂ cells, γ-secretase inhibitor, arsenic trioxide, apoptosis

中图分类号:

R735.7

周明, 王雪雯, 杨莉, 宋海林, 段晶晶. As₂O₃联合γ分泌酶抑制剂对HepG₂细胞凋亡的诱导作用及其机制[J]. 吉林大学学报(医学版), 2015, 41(03): 470-475.

ZHOU Ming, WANG Xuewen, YANG Li, SONG Hailin, DUAN Jingjing. Induction of As₂O₃ in combination with γ-secretase inhibitor on apoptosis of HepG₂ and its mechanism[J]. Journal of Jilin University Medicine Edition, 2015, 41(03): 470-475.

参考文献

[1] Stewart BW.Wild CP[R].World Cancer Report, 2014.

[2] Li W, Wang M, Wang L, et al.Icariin synergizes with arsenic trioxide to suppress human hepatocellular carcinoma [J].Cell Biochem Biophys, 2014, 68 (2):427-436.

[3] Liu XM, Xiong XF, Song Y, et al.Possible roles of a tumor suppressor gene PIG11 in hepatocarcinogenesis and As₂O₃-induced apoptosis in liver cancer cells [J].J Gastroenterol, 2009, 44(5):460-469.

[4] Huang X, Ma J, Xu J, et al.Simvastatin induces growth inhibition and apoptosis in HepG₂ and Huh7 hepatocellular carcinoma cells via upregulation of Notch1 expression [J].Mol Med Rep, 2014, 11(3): 2334-2340.

[5] Li D, Masiero M, Banham AH, et al.The notch ligand JAGGED1 as a target for anti-tumor therapy [J].Front Oncol, 2014, 25(4):254-263.

[6] Yabuuchi S, Pai SG, Campbell NR, et al.Notch signaling pathway targeted therapy suppresses tumor progression and metastatic spread in pancreatic cancer [J].Cancer Lett, 2013, 335(1): 41-51.

[7] Rasul S, Balasubramanian R, Filipovic A, et al.Inhibition of gamma-secretase induces G₂/M arrest and triggers apoptosis in breast cancer cells [J].Br J Cancer.2009, 100(12):1879-1888.

[8] Timme CR, Gruidl M, Yeatman TJ, et al.Gamma-secretase inhibition attenuates oxaliplatin-induced apoptosis through increased Mcl-1 and/or Bcl-xL in human colon cancer cells [J].Apoptosis, 2013, 18(10):1163-1174.

[9] Gao J, Dong Y, Zhang B, et al.Notch1 activation contributes to tumor cell growth and proliferation in human hepatocellular carcinoma HepG₂ and SMMC7721 cells [J].International J Oncol, 2012, 41(5): 1773-1781.

[10] Wang Z, Banerjee S, Li Y, et al.Down-regulation of Notch-1 inhibits invasion by inactivation of nuclear factor-κB, vascular endothelial growth factor, and matrix metalloproteinase-9 in pancreatic cancer cells[J].Cancer Res, 2006, 66(5): 2778-2784.

[11] Qi R, An H, Yu Y, et al.Notch1 signaling inhibits growth of human hepatocellular carcinoma through induction of cell cycle arrest and apoptosis[J].Cancer Res, 2003, 63(23): 8323-8329.

[12] Wang C, Qi R, Li N, et al.Notch1 signaling sensitizes tumor necrosis factor-related apoptosis-inducing ligand-induced apoptosis in human hepatocellular carcinoma cells by inhibiting Akt/Hdm2-mediated p53 degradation and up-regulating p53-dependent DR5 expression[J].Biol Chem, 2009, 284(24): 16183-16190.

[13] Wang WZ, Li L, Liu MY, et al.Curcumin induces FasL-related apoptosis through p38 activation in human hepatocellular carcinoma Huh7 cells [J].Life Sci, 2013, 92(6):352-358.

[13] Zhang YQ, Xiao CX, Lin BY, et al.Silencing of Pokemon enhances caspase-dependent apoptosis via fas- and mitochondria-mediated pathways in hepatocellular carcinoma cells [J].PLoS One, 2013, 8(7):681-689.

[14] Chen GI, Wang K, Yang BY, et al.Synergistic antitumor activity of oridonin and arsenic trioxide on hepatocellular carcinoma cells [J].Int Oncol, 2012, 40(1): 139-147.

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As₂O₃联合γ分泌酶抑制剂对HepG₂细胞凋亡的诱导作用及其机制

Induction of As₂O₃ in combination with γ-secretase inhibitor on apoptosis of HepG₂ and its mechanism

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