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目的探讨RNA干扰结肠癌相关基因BC047440的表达对结肠癌细胞HCT-8增殖能力的影响。方法将BC047440基因shRNA干扰质粒pGPU6/GFP/Neo-BC047440-331(a1)、pGPU6/GFP/Neo-BC047440-451(a2)、pGPU6/GFP/Neo-BC047440-615(a3)、pGPU6/GFP/Neo-BC047440-756(a4)采用脂质体法转染结肠癌细胞HCT-8,荧光显微镜观察转染效率,荧光定量PCR法检测细胞BC047440基因mRNA的表达水平,MTT法分析细胞的增殖活性。结果细胞的转染效率为53%;干扰质粒a1、a2、a3、a4转染的HCT-8细胞BC047440基因mRNA的表达水平分别下降了36.1%、47.0%、53.8%和63.3%,a4质粒的干扰效率最高;细胞的增殖能力也明显下降。结论 BC047440基因RNA干扰质粒可明显抑制HCT-8细胞BC047440基因mRNA的表达及细胞的增殖,可能成为抑制结肠癌细胞生长的新基因。
Objective To investigate the effect of RNA interference of colon cancer related gene BC047440 on the proliferation of human colon cancer cell line HCT-8. Methods BC047440 gene shRNA interference plasmid pGPU6 / GFP / Neo-BC047440-331 (a1), pGPU6 / GFP / Neo-BC047440-451 (a2), pGPU6 / GFP / Neo-BC047440-615 (a3), pGPU6 / GFP / Neo-BC047440-756 (a4) The colon cancer cell line HCT-8 was transfected by lipofectamine. The transfection efficiency was observed by fluorescence microscopy. The expression of BC047440 gene mRNA was detected by real-time quantitative PCR. The proliferation activity was analyzed by MTT assay. Results The transfection efficiency of BC047440 gene in HCT-8 cells transfected with plasmids a1, a2, a3 and a4 were decreased by 36.1%, 47.0%, 53.8% and 63.3%, respectively. The transfection efficiency of a4 plasmid The highest interference efficiency; cell proliferation also decreased significantly. Conclusion BC047440 gene RNAi plasmid can significantly inhibit HCT-8 cells BC047440 mRNA expression and cell proliferation, may be a new gene that inhibits the growth of colon cancer cells.