shRNA-聚束蛋白基因对人结肠癌细胞SW-480侵袭和凋亡影响的实验研究

Experimental study of the invasion and apoptosis of shRNA-Fascin gene on human colon cancer SW-480 cells

目的探究shRNA-Fascin基因对人结肠癌细胞SW-480侵袭和凋亡影响。方法利用慢病毒转染技术,构建Fascin的shRNA干扰表达载体,RT-qPCR和Western blot等方法检测Fascin基因和凋亡蛋白Caspase-3和Caspase-9的表达。Transwell试剂盒检测人结肠癌细胞的侵袭,以及AV-PI试剂盒检测人结肠癌细胞的凋亡。结果shRNA-Fascin载体慢病毒转染效率为95%以上。实验组SW-480中Caspase-3和Caspase-9的mRNA的相对表达量明显高于空白对照组,差异有统计学意义(P<0.05)。实验组SW-480中Caspase-3和Caspase-9的蛋白相对表达水平明显高于空白对照组,差异有统计学意义(P<0.05)。实验组SW-480细胞侵袭能力明显低于空白对照组,差异有统计学意义(P<0.05)。实验组SW-480细胞的凋亡率明显高于空白对照组,差异有统计学意义(P<0.05)。结论shRNA-Fascin可以明显的抑制人结肠癌细胞SW-480的侵袭,促进人结肠癌细胞SW-480的凋亡。

Objective To investigate the effect of shRNA-Fascin gene on the invasion and apoptosis of human colon cancer SW-480 cells.Methods Fascin shRNA interference expression vector was constructed by lentivirus transfection.The expressions of Fascin and apoptotic proteins Caspase-3 and Caspase-9 were detected by RT-qPCR and Western blot.Transwell kit was used to detect invasion of human colon cancer cells,and AV-PI kit was used to detect apoptosis of human colon cancer cells.Results The shRNA-Fascin vector lentivirus transfection efficiency is above 95%.The relative expressions of Caspase-3 and Caspase-9 mRNA in SW-480 in experimental group were significantly higher than that in blank control group,with the differences statistically significant(P<0.05).The relative levels of Caspase-3 and Caspase-9 in SW-480 in experimental group were significantly higher than those in blank control group,with the differences statistically significant(P<0.05).The invasion ability of SW-480 cells in the experimental group was significantly lower than that in the blank control group,with the differences statistically significant(P<0.05).The apoptosis rate of SW-480 cells in the experimental group was significantly higher than that in the blank control group,with the differences statistically significant(P<0.05).Conclusion shRNA-Fascin can significantly inhibit the invasion of human colon cancer cell SW-480 and promote the apoptosis of human colon cancer cell SW-480.