摘要
目的:观察依托泊苷(etoposide,VP-16)对结肠癌细胞衰老的影响。方法:0～8μg/mL VP-16作用结肠癌LS-174T细胞后,采用CCK-8(cell counting kit-8,CCK-8)法检测细胞增殖情况,FCM检测细胞凋亡及细胞周期改变,衰老相关β-半乳糖苷酶(senescence-associated β-galactosidase,SA-β-gal)染色检测细胞衰老情况,Western印迹法检测p53、p16、RB、p21WAF-1/CIP1和E2F1蛋白的表达。结果:VP-16可以抑制结肠癌LS-174T细胞的增殖,且呈浓度依赖性;大剂量(2和4μg/mL)VP-16可以促进结肠癌细胞凋亡;小剂量(1μg/mL)VP-16作用后,结肠癌细胞呈现典型的大而扁平的衰老形态,SA-β-gal染色阳性,细胞周期阻滞于G2/M期。Western印迹法检测结果显示,小剂量VP-16可以促进p21WAF-1/CIP1蛋白表达,抑制E2F1蛋白表达,而对p53、p16和RB蛋白表达无影响。结论:大剂量VP-16可促进结肠癌细胞凋亡;小剂量VP-16可促进结肠癌细胞衰老,其作用机制可能与VP-16促进细胞p21WAF-1/CIP1蛋白表达及抑制E2F1蛋白表达有关。
Objective：To observe the effects of etoposide on cell senescence in colon carcinoma.Methods： Colon carcinoma LS-174T cells were exposed to various doses of etoposide（0-8 μg/mL）.Cell proliferation was detected with cell counting kit-8（CCK-8） assay.Cell apoptosis and cell cycle distribution were detected by flow cytometry.Cell senescence was detected by senescence-associated β-galactosidase（SA-β-gal） staining.The expressions of p53,p16,RB,p21WAF-1/CIP1,and E2F1 protein were detected by Western blotting.Results：Etoposide inhibited proliferation of colon carcinoma cells in a dose-dependent manner.High doses of etoposide（2 and 4 μg/mL） could induce apoptosis in colon carcinoma cells.Colon carcinoma cells exhibited typical enlarged and flattened senescence morphology after treatment with 1 μg/mL etoposide.The cells showed positive SA-β-gal staining and were arrested in G2/M phase.Western blotting showed that low dose of etoposide up-regulated expression of p21WAF-1/CIP1 and down-regulated E2...更多F1 protein expression.Conclusion： High doses of etoposide could induce apoptosis of colon carcinoma cells.Low dose of etoposide could induce senescence in colon carcinoma cells and may related to up-regulation of p21WAF-1/CIP1 and down-regulation of E2F1 protein.
出处
《肿瘤》
CAS
CSCD
北大核心
2009年第8期711-714,共4页
Tumor
基金
上海市自然科学基金资助项目(编号:07ZR14109)
上海市教育委员会科研项目(编号:08cz018)
上海市市级医院临床科研资源共享平台资助项目(编号:SHDC12007206)
