MAP4K4沉默表达对结肠癌细胞生物学特性的影响

Effects of silenced MAP4K4 expression on biological features of colon cancer cells

目的分析沉默丝裂原活化蛋白激酶-激酶激酶4(MAP4K4)对结肠癌细胞生物学行为的影响。方法脂质体siRNA干扰技术转染人结肠癌细胞SW620细胞株,按转染情况分为MAP4K4干扰组(转染MAP4K4-siRNA 1.5 mL)、空载体组(转染无关siRNA 1.5 mL)、空白对照组(不予细胞转染处理)。分别采用逆转录-聚合酶链反应(RT-PCR)和蛋白印迹法(WB)测定各组MAP4K4 mRNA及蛋白表达,并检测各组SW620细胞增殖、侵袭及细胞凋亡情况。结果①空载体组与MAP4K4干扰组MAP4K4 mRNA、蛋白浓度低于空白对照组(P<0.05),MAP4K4干扰组MAP4K4 mRNA、蛋白浓度低于空载体组(P<0.05);②MAP4K4干扰组不同时间增殖活性均低于空载体组和空白对照组(P<0.05);③MAP4K4干扰组穿膜细胞比率低于空载组和空白对照组(P<0.05),空载体组穿膜细胞比率低于空白对照组(P<0.05);④MAP4K4干扰组细胞凋亡率高于空载组和空白对照组(P<0.05),空载体组细胞凋亡率高于空白对照组(P<0.05)。结论siRNA干扰技术沉默结肠癌细胞MAP4K4表达可降低结肠癌细胞增殖及侵袭能力,促进癌细胞凋亡,或可能为结肠癌防治的新靶点。

Objective To analyze the effects of silenced mitogen activated protein kinase kinase 4(MAP4K4)on biological behaviors of colon cancer cells.Methods Human colon cancer cell lines SW620 were transfected with liposome siRNA interference technique.According to the condition of transfection,they were divided into MAP4K4 interference group(transfection with 1.5 mL of MAP4K4-siRNA),empty vector group(transfection with 1.5 mL of unrelated siRNA)and blank control group(no transfection).The mRNA and protein expression of MAP4K4 was determined.The proliferation,invasion and apoptosis of SW620 cells in each group were detected.Results①The MAP4K4 mRNA and protein concentrations in the empty vector group and MAP4K4 interference group was lower than in the blank control group(P<0.05).②The proliferation activity in MAP4K4 interference group was lower than that in empty vector group and blank control group at different time(P<0.05).③The order of the ratio of transmembrane cells from low to high was as follows:MAP4K4 interference group,empty vector group,blank control group(P<0.05).④The order of apoptosis rate from high to low was as follows:MAP4K4 interference group,empty vector group,blank control group(P<0.05).Conclusion Using the SiRNA interference technique to silence the expression of MAP4K4 in colon cancer cells can reduce the proliferation and invasion ability of colon cancer cells,and promote their apoptosis.It may be a new target for controlling colon cancer.