特异性沉默EPB49对结直肠癌细胞转移能力的影响以及作用机制

The effect of specific silencing EPB49 on cell metastasis of colorectal cancer and its mechanism

目的:研究小干扰RNA(small interfering RNA,siRNA)特异性沉默红细胞膜蛋白带4. 9(erthrocyte brane protein band 4. 9,EPB49)对结直肠癌SW480细胞增殖、侵袭、迁移的影响以及可能的作用机制。方法:将结直肠癌SW480细胞分为空白对照组、NC对照组(转染阴性对照siRNA-NC)、siRNA-EPB49组,通过Lipofectamine 2000将siRNA-NC和siRNA-EPB49转染入SW480细胞; CCK-8实验检测细胞的增殖活力;Transwell法检测SW480细胞侵袭、迁移能力的变化;蛋白质印迹法(Western Blot)检测EPB49、信号转导与转录因子3 (signal transduction and transcription factor 3,STAT3)、磷酸化STAT3 (phosphorylated STAT3,pSTAT3)、另一类激酶2(just another kinase 2,JAK2)、磷酸化JAK2(p-JAK2)蛋白的表达。结果:沉默EPB49表达后,与空白对照组相比,siRNA-EPB49组细胞中EPB49蛋白的表达量显著降低(P <0. 05),细胞增殖活性显著增加(P <0. 05),侵袭、迁移能力显著增加(P <0. 05); Western Blot检测结果表明特异性沉默EPB49后SW480细胞中p-STAT3、p-JAK2蛋白表达量明显增加(P <0. 05)。结论:沉默EPB49可促进结直肠癌细胞增殖、侵袭、迁移,此作用与JAK2/STAT3信号通路有关。

Objective:To investigate the effect of small interfering RNA(siRNA)silencing erthrocyte brane protein band 4.9(EPB49)on proliferation,invasion and migration of colorectal cancer SW480 cells and its possible mechanism.Methods:Colorectal cancer SW480 cells were divided into blank control group,NC control group(transfected negative control siRNA-NC),siRNA-EPB49 group.siRNA-NC and siRNA-EPB49 were transfected into SW480 cells by Lipofectamine 2000.The proliferation activity of the cells was detected by CCK-8 experiment.Transwell assay was used to detect the invasion and migration of SW480 cells.Western Blot was used to detect the expression of EPB49,signal transduction and transcription factor 3(STAT3),phosphorylated STAT3(p-STAT3),just another kinase 2(JAK2),and phosphated JAK2(p-JAK2)protein.Results:After silencing EPB49 expression,the expression of EPB49 protein in siRNA-EPB49 group was significantly decreased than that in blank control group(P<0.05),and the cell proliferation activity,invasion and migration ability increased significantly(P<0.05).The results of Western Blot showed that the expression of p-STAT3 and p-JAK2 in SW480 cells increased significantly after EPB49 silenced(P<0.05).Conclusion:Silencing EPB49 can promote the proliferation,invasion and migration of colorectal cancer cells,which is related to the JAK2/STAT3 signaling pathway.