蛋白酶體抑制劑MG132對膽囊癌細胞殺傷作用的實驗研究

朱衛平，詹棣華，趙一鳴，王魯，解宏偉，王會鵬，程志儉（.復旦大學附屬上海市第五人民醫院　普外科，上海　0040；.復旦大學附屬腫瘤醫院　肝臟外科，上海0003）

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蛋白酶體抑制劑MG132對膽囊癌細胞殺傷作用的實驗研究

朱衛平1，2，詹棣華2，趙一鳴2，王魯2，解宏偉1，王會鵬1，程志儉1
（1.復旦大學附屬上海市第五人民醫院普外科，上海200240；2.復旦大學附屬腫瘤醫院肝臟外科，上海200032）

[摘 要]目的 研究蛋白酶體抑制劑MG132抑制膽囊癌細胞增殖及誘導凋亡的機制。方法 采用CCK8和流式細胞術檢測膽囊癌細胞增殖及凋亡情況；采用Western blotting和RT-PCR檢測凋亡相關基因Caspase-8、Caspase-3和DR5的mRNA和蛋白質表達；建立荷瘤裸鼠模型，觀察MG132干預下裸鼠瘤重及瘤體積的變化。結果 在本研究中發現MG132能有效抑制體外和體內膽囊癌細胞的增殖，其作用呈劑量依賴性（P＜0.01）。MG132能促使膽囊癌細胞發生G2/M期阻滯及誘導細胞凋亡。MG132誘導膽囊癌細胞凋亡主要通過激活外源性凋亡通路中DR5、Caspase-8和Caspase-3過表達。結論 MG132能對膽囊癌細胞起殺傷作用，其機制可能通過影響細胞周期阻滯及誘導細胞凋亡起作用。

[關鍵詞]膽囊癌；蛋白酶體抑制劑；增殖；凋亡

Experimental study of the killing effect of proteasome inhibitor MG132 on gallbladder cancer cells

ZHU Wei-ping1,2,ZHAN Di-hua2,ZHAO Yi-ming2,Wang Lu2,Xie Hong-wei1,Wang Hui-peng1,Cheng Zhi-jian1.1Department of General Sugery,the Fifth People’s Hospital of Shanghai,Fudan University,Shanghai 200240,China;2Department of Hepatic Surgery,Fudan University Shanghai Cancer Center,Shanghai,200032,China

Abstract objectiveTo investigate the mechanism of proteasome inhibitor MG132 in inhibiting proliferation and inducing apoptosis of gallbladder cancer cells.MethodsThe anti-tumor activity of MG132 on gallbladder cancer cells was assessed by the CCK8 assay.Apoptosis changes were detected by flow cytometric analysis.The protein and mRNA expression of Caspase-8,Caspase-3 and DR5 were examined using Western blotting and RT-PCR.Gallbladder cancer xenografts in athymic nude mice were established and the tumor weight and volume were measured after the intervention of MG132.ResultsThe results showed that MG132 inhibited the proliferation of gallbladder cancer cells in a dose-dependent manner both in vivo and in vitro(P＜0.01).Exposure of tumor cells to MG132 induced cell cycle arrest at G2/M phase and apoptosis in a dose- dependent manner(P＜0.01).In addition,analysis of apoptotic pathways indicated that MG132 significantly enhanced the activation of Caspase-8,Caspase-3 and DR5 in extrinsic signaling pathway(P＜0.05).ConclusionProteasome inhibitor MG132 can effectively kill gallbladder cancer cells and its mechanism involves the cell cycle arrest and apoptosis induction.

Key words gallbladder carcinoma; proteasome inhibitor; proliferation; apoptosis

膽囊癌是高度惡性的膽道系統腫瘤，目前雖然隨著診斷技術和治療手段的提高使得患者預后有所改善，但5年生存率仍然低于5%[1]。因此，積極探索膽囊癌治療的新策略成為目前研究的重點。泛素-蛋白酶體通路（ubiquitin-proteasome pathway，UPP）是調控真核細胞中內源性蛋白質選擇性降解的關鍵途徑，與腫瘤細胞的周期運轉、增殖、凋亡等生物學行為密切相關[2]。……