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RNA interference inhibits the proliferation of human colorectal cancer cells by blocking the expression of cyclin D1

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Author:
No author available
Journal Title:
WORLD CHINESE JOURNAL OF DIGESTOLOGY
Issue:
34
DOI:
10.3969/j.issn.1009-3079.2007.34.003
Key Word:
小干扰RNA;结肠癌;cyclin D1;细胞周期;细胞增殖;MTT比色法;流式细胞技术

Abstract´╝Ü 目的:使用RNA干扰技术阻断人结肠癌细胞cyclin D1的表达,检测其对细胞生长增殖的影响和机制.方法:将人结肠癌细胞株HT-29分成3组,antisense组(转染反义链组)、sense组(转染正义链组)和对照组.免疫沉淀和蛋白质印迹法观察cyclin D1 siRNA对细胞cyclin D1蛋白质表达的影响,并对蛋白质电泳图像进行分析,MTT比色法绘制细胞生长曲线,3H-TdR技术和流式细胞技术观测细胞周期变化.结果:反义siRNA有效抑制了cyclin D1蛋白质表达,MTT实验显示,转染反义siRNA的细胞,生长代谢减慢,与对照组细胞差异显著(P<0.01).而antisense组细胞24 h的3H-TdR渗入量,G0/G1期和S期细胞较对照组和sense组明显减少(3H-TdR:1181.8±117.97 vs 1798.4±55.36,1851.4±83.46;P<0.01;G0/G1期:79.31%vs 60.87%,59.14%;S期:13.67% vs 26.42%,27.93%;P<0.01).结论:RNA干扰技术能有效减弱cyclin D1蛋白质表达,使细胞周期受阻,并抑制结肠癌细胞的生长增殖.

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