RNA干扰HER2基因对人大肠癌HT29细胞株增殖影响的研究
Inhibitory Effects of HER2 RNAi on Proliferation of HT29 Cells
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摘要: 目的研究载体介导的靶向HER2基因的RNA干扰 (RNA interference,RNAi)对体外生长的人大肠癌HT29细胞增殖的影响。方法 将针对HER2基因的短发夹RNA(short hairpin RNA,shRNA)表达载体转染体外生长的HT29细胞,分别用半定量RT-PCR和Western blot检测HER2 mRNA和蛋白的表达,MTT测定各组细胞的增殖差异。结果 靶向HER2的shRNA可以有效地抑制体外生长的人大肠癌HT29细胞中HER2基因mRNA和蛋白表达,与对照组比较,实验组细胞增殖明显受到抑制。结论 靶向HER2的shRNA表达载体可以通过降低HER2基因的表达,进而抑制体外生长的大肠癌HT29细胞的增殖。Abstract: ObjectiveTo investigate the inhibitory effect of RNAi targeting HER2 on the proliferation of human colorectal cancer cells in vitro. Methods Human colorectal cancer HT-29 cells were transfected with plasmid expressing shRNA-HER2 by LipofectamineTM2000. The mRNA level and expression of HER2 in HT29 cells of each group were detected using semi-quantitive reverse transcription-polymerase chain reaction(RT-PCR) and Western blot,respectively. Inhibitory effect on proliferation of each group was measured by MTT assay. Results Plasmid mediated shRNA-HER2 can depress both transcriptional and translational level of HER2 in HT29 cells in vitro. Compared with control the proliferation of HT-29 cells was inhibited by shRNA-HER2 (P<0.01). Conclusion Plasmid mediated shRNA-HER2 can inhibit the proliferation of HT-29 cells by reducing the HER2 gene expression in vitro.