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Lsm4mRNA的表达。将靶向Lsm4基因的小干扰I矾A(smallinterferenceRNA,siRNA)(Lsm4-siRNA)或阴性对照(negativecontrol,NC)(NC—siRNA)转染入食管癌ECl09细胞,实时荧光定量-PCR法检测细胞中Lsm4mRNA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(proliferatingcellnuclearantigen,PCNA)和波形蛋白的表达水平。结果:食管癌组织中Lsm4mRNA的表达水平明显高于相应的癌旁组织(P〈0.05)。Lsm4-siRNA转染后,ECl09细胞中Lsm4mRNA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ECl09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PC烈A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Lsm4mI埘A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.