E47和LOXL2表达下调对食管鳞癌Eca-109细胞迁移和凋亡的影响研究

Effect of down-regulated E47 and LOXL2 expression on Eca-109 cell migration and apoptosis in esophageal squamous cell carcinoma

目的探讨核转录因子E47和赖氨酰氧化酶样蛋白-2(LOXL2)基因表达下调后对食管鳞癌细胞株Eca109迁移、凋亡的影响及其机制。方法采用RNAi技术沉默E47和LOXL2基因作为实验组,同时设立空白对照组和阴性对照组。采用实时荧光定量PCR及Western blot检测各组细胞中E47、LOXL2基因mRNA和蛋白表达水平,通过流式细胞术和Transwell检测细胞凋亡和迁移变化,采用免疫共沉淀技术检测E47和LOXL2蛋白相互作用。结果与空白对照组和阴性对照组比较,E47、LOXL2 mRNA和蛋白表达水平下调;流式细胞术和Transwell实验结果显示,E47和LOXL2干扰后Eca109细胞的凋亡率增高(P<0.01),细胞迁移数目降低(P<0.01)。免疫共沉淀结果显示E47和LOXL2蛋白存在相互作用。结论E47和LOXL2表达下调可以抑制Eca109细胞迁移,并诱导细胞凋亡,E47和LOXL2蛋白相互作用可能促进食管癌细胞的恶性增殖和侵袭。

Objective To explore the effect of Nuclear transcription factor E47 and lysyl oxidase-like 2(LOXL2)genes silencing on the cell migration and apoptosis in human esophagus squamouscell cancer Eca109 cells and relevant mechanism.Methods E47 and LOXL2 genes were silenced by RNAi technology and used as the experimental group,meanwhile the blank control group and negative control group were set up.The mRNA and protein expression levels were detected by real-time fluorogenic quantitative PCR and Western blot individually.Flow cytometry and transwell chamber assay were used to examine the apoptosis and migration of Eca109 cells.Co-immunoprecipitation technique was employed for analysis of the relationship of them.Results Compared with those of blank and negative control groups,mRNA and protein levels of E47 and LOXL2 were lower in the experimental group;the apoptosis were promoted(P<0.01)and cell migration ability were reduced(P<0.01)of Eca109 cells.The results of co-immunoprecipitation showed that there was an interaction between the E47 and LOXL2 proteins.Conclusion shE47 and shLOXL2 induce apoptosis and suppress the abilities of migration in Eca109 cells.There is an interaction between the E47 and LOXL2 protein,which may promote the malignant proliferation and invasion of esophageal cancer cells.