文档介绍：白藜芦醇对大肠癌细胞hTERT表达及其启动子的影响
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作者:王晓燕, 范钰, 张尤历, 钟锡明
【摘要】目的: 观察白藜芦醇对大肠癌细胞生长增殖的影响,并从人端粒酶逆转录酶(human telomerase reverse transcriptase,hTERT)表达及其启动子方面探讨可能的抗癌机制。方法: 以不同浓度的白藜芦醇处理大肠癌LS174T细胞后,收集细胞,分别采用荧光实时定量RTPCR和蛋白质印迹法检测癌细胞hTERT mRNA和蛋白水平。采用脂质体转染法将携带hTERT启动子和报告基因的质粒转染至大肠癌LS174T细胞中后,加入不同浓度的白藜芦醇,孵育48 h后,检测报告基因萤虫素酶活性。结果: 白藜芦醇处理大肠癌细胞后,增殖明显受到抑制,且与浓度有关。癌细胞hTERT mRNA和蛋白水平明显下调,且呈浓度和时间依赖性。白藜芦醇处理组萤虫素酶活性下降,且呈浓度依赖性。结论: hTERT启动子活性和hTERT表达下调,可能是白藜芦醇抑制癌细胞增殖的重要机制之一。
【关键词】白藜芦醇; 大肠肿瘤; 人端粒酶逆转录酶; 启动子
[Abstract] Objective: To investigate the effect of resveratrol on human telomerase reverse transcriptase(hTERT) and its promoter of human colorectal cancer cell. Methods: After LS174T human colorectal cancer cell were treated with different dose of resveratrol,mRNA and protein of hTERT were determined by realtime PCR and Western blot,respectively. The plasmid inserted hTERT promoter was transected into LS174T cells by oligofecamine 2000, and different doses of resveratrol were added into culture media 2 hours later. After 48 hours, luciferase activity of colorectal cancer cell was determined. Results: The expression of mRNA and protein of hTERT of cells treated with resveratrol were downregulated in doseand timedependent manner. The luciferase activity of cancer cells treated with resveratrol were inhibited in dosedependent manner. Conclusion: Resver