miR-137下调FABP4抑制食管腺癌细胞的增殖、迁移的实验研究

Experimental study on the inhibition of proliferation and migration of esophageal adenocarcinoma cells by down-regulation of FABP4 by miR-137

目的探讨miR-137通过下调脂肪酸结合蛋白4(FABP4)表达对食管腺癌细胞增殖、迁移影响及可能机制。方法采用实时荧光定量PCR(q PCR)检测食管腺癌BIC-1、SEG1细胞和人正常食管上皮细胞HET-1A中FABP4和miR-137的表达。分别向SEG1细胞转染NC-mimics和pc DNA3. 1-vector(阴性对照组)、miR-137 mimics(miR-137组)、miR-137 mimics和pc DNA3. 1-vector(miR-137-NC组)和miR-137 mimics和pc DNA3. 1-FABP4(miR-137+FABP4组),另设置不作任何处理的空白对照组。采用MTT法、Transwell小室法检测各组细胞增殖和迁移的活性。结果与HET-1A细胞比较,BIC-1和SEG1细胞中FABP4(1. 00±0. 07 vs. 1. 43±0. 07 vs. 1. 64±0. 09)表达升高,miR-137表达降低(1. 00±0. 04 vs. 0. 76±0. 06 vs. 0. 51±0. 04),差异有统计学意义(P<0. 05)。Western blotting法检测结果显示,miR-137组中FABP4蛋白表达水平低于空白对照组、阴性对照组(0. 35±0. 04 vs. 0. 48±0. 06 vs. 0. 44±0. 07),差异有统计学意义(P<0. 05)。MTT法检测结果显示,miR-137组细胞增殖活性低于空白对照组和阴性对照组(P<0. 05);miR-137+FABP4组细胞增殖活性高于miR-137组(P<0. 05)。Transwell小室迁移实验结果显示,miR-137组细胞迁移活性低于空白对照组和阴性对照组(P<0. 05);miR-137+FABP4组细胞迁移活性高于miR-137组(P<0. 05)。结论 miR-137通过下调FABP4表达抑制食管腺癌细胞增殖和迁移活性。

Objective To investigate the role and possible mechanism of miR-137 in inhibiting proliferation and migration of esophageal adenocarcinoma cells by down-regulating the expression of fatty acid binding protein 4(FABP4).Methods The expressions of FABP4 and miR-137 in BIC-1 and SEG1 cells and human normal esophageal epithelial cells HET-1A were detected by real-time fluorescent quantitative PCR(qPCR).SEG1 cells were transfected with NC-mimics and pcDNA3.1-vector(negative control group),miR-137 mimics(miR-137 group),miR-137 mimics and pcDNA3.1-vector(miR-137-NC group)and miR-137 mimics and pcDNA3.1-FABP4(miR-137+FABP4 group),respectively,and blank control group was set without any treatment.The activity of cell proliferation and migration was detected by MTT and Transwell assay.Results Compared with HET-1A cells,the expression of FABP4 was increased in BIC-1 and SEG1 cells and the expression of miR-137 was decreased in BIC-1 and SEG1 cells,with statistically significant differences(P<0.05).Western blotting showed that the expression level of FABP4 protein in the miR-137 group was lower than that in the blank control group and the negative control group,and the difference was statistically significant(P<0.05).The results of MTT assay showed that the cell proliferation activity of miR-137 group was lower than that of blank control group and negative control group(P<0.05).The cell proliferation activity of miR-137+FABP4 group was higher than that of miR-137 group(P<0.05).The results of Transwell chamber migration experiment showed that the cell migration activity of miR-137 group was lower than that of the blank control group and the negative control group(P<0.05).The cell migration activity of miR-137+FABP4 group was higher than that of miR-137 group(P<0.05).Conclusion miR-137 inhibited proliferation and migration activity of esophageal adenocarcinoma cells by down-regulating the expression of FABP4.