RNA结合蛋白Lsm4对食管癌EC109细胞增殖和迁移的影响

The effect of RNA binding protein Lsm4 on the proliferation and migration of esophageal carcinoma cell line EC109

目的 :探讨干扰Lsm4基因对人食管癌EC109细胞增殖和迁移能力的影响及其可能的机制。方法 :实时荧光定量-PCR法检测26例食管癌及其相应癌旁组织中Lsm4 mRNA的表达。将靶向Lsm4基因的小干扰RNA(small interference RNA,siRNA)(Lsm4-siRNA)或阴性对照(negative control,NC)(NC-siRNA)转染入食管癌EC109细胞,实时荧光定量-PCR法检测细胞中Lsm4 mRNA的表达,MTT法和平板克隆形成实验检测细胞的增殖和克隆形成能力,Transwell法检测细胞的迁移能力,蛋白质印迹法检测Lsm4、细胞核增殖抗原(proliferating cell nuclear antigen,PCNA)和波形蛋白的表达水平。结果 :食管癌组织中Lsm4 mRNA的表达水平明显高于相应的癌旁组织(P<0.05)。Lsm4-siRNA转染后,EC109细胞中Lsm4 mRNA及其蛋白的表达水平低于阴性对照组,差异有统计学意义(P<0.01);Lsm4-siRNA转染后,EC109细胞的增殖和克隆形成能力受到抑制(P<0.05),而EC109细胞的迁移能力增强(P<0.01)。与阴性对照组比较,Lsm4-siRNA转染组细胞PCNA的表达水平下调(P<0.05),而波形蛋白的表达水平上调(P<0.01)。结论 :食管癌组织中Lsm4 mRNA高表达,Lsm4可能参与调控食管癌EC109细胞的增殖和迁移。

Objective： To investigate the effect of Ism4 gene interference on the proliferation and migration of esophageal carcinoma EC109 cells, and to explore its possible mechanism. Methods： The expression of Lsm4 mRNA in esophageal cancer tissues and para-carcinoma tissues was detected by real-time fluorescence quantitative-PCR （RFQ-PCR）. After transfection with Lsm4-small interference RNA （siRNA） targeting human Isrn4 gene or a negative control （NC） （NC-siRNA）, the expression level of Lsm4 mRNA in EC109 cells was detected by RFQ-PCR, and the abilities of cell proliferation and colony formation were determined by MTT and colony formation assay, respectively. The ability of migration of EC109 cells after transfection with Lsm4-siRNA was examined by Transwell assay, and the expression levels of Lsm4, proliferating cell nuclear antigen （PCNA） and vimentin proteins were detected by Western blotting. Results： The expression level of Lsm4 mRNA in esophageal cancer tissues was higher than that in para-carcinoma tissues （P 〈 0.05）. The expression levels of Lsm4 mRNA and protein of EC109 cells in Lsm4-siRNA transfection group were lower than those in NC-siRNA group （P 〈 0.01）. The abilities of proliferation and colony formation of EC109 cells in Lsm4-siRNA transfection group were inhibited （P 〈 0.05）, while the ability of migration was improved （P 〈 0.01）. As compared with NC group, the expression level of PCNA was down-regulated （P 〈 0.05）, and the expression level of vimentin was up-regulated （P 〈 0.01）. Conclusion： The expression of Lsm4 mRNA is higher in esophageal carcinoma tissues. Lsm4 may regulate the growth and migration of EC109 cells.