miR-30-5p在食管癌中的表达及对食管癌细胞增殖和迁移的影响

Expression of miR-30-5p in esophageal cancer and its effect on proliferation and migration of esophageal cancer cells

目的:探讨miR-30-5p对食管癌细胞增殖和迁移的影响及分子机制。方法:选取开封市中心医院心胸血管外科和河南省人民医院胸外科2014年6月到2019年6月收集的食管癌和癌旁组织样本各120例,采用荧光定量PCR测定miR-30-5p mRNA表达水平。采用慢病毒建立食管癌细胞系EC109中建立miR-30-5p过表达稳定细胞系(miR-30-5p组)和对照细胞系(miR-control组)。采用EdU染色分析2组细胞的增殖情况;采用Transwell分析2组细胞的迁移情况;采用生物信息学和双荧光素酶报告基因分析miR-30-5p的靶基因;在miR-30-5p组和miR-control组过表达靶基因对细胞增殖和迁移的影响。结果:与癌旁组织比较,食管癌组织中miR-30-5p mRNA表达水平明显上调,差异有统计学意义(P<0.05)。与miR-control组比较,miR-30-5p组细胞增殖能力和细胞迁移能力明显下降,差异有统计学意义(P<0.05)。生物信息学和双荧光素酶报告基因显示BCL-9是miR-30-5p的靶基因。在miR-control组细胞中过表达BCL-9,可显著提高细胞的增殖和迁移,差异有统计学意义(P<0.05)。而miR-30-5p组细胞过表达BCL-9,细胞增殖和迁移差异无统计学意义(P<0.05)。结论:miR-30-5p在食管癌中呈低表达,可导致靶基因BCL-9表达上调,进而促进食管癌细胞的增殖和迁移。

Objective:To investigate the effect of miR-30-5 p on the proliferation and migration of esophageal cancer cells and its molecular mechanism. Methods:A total of 120 samples of esophageal cancer and 120 samples of adjacent tissues were collected in department of cardiothoracic vascular surgery of Kaifeng Central Hospital and thoracic surgery of Henan People’s Hospital from June2014 to June 2019,and fluorescence quantitative PCR was used to detect the expression of miR-30-5 p. Lentivirus was used to establish miR-30-5 p overexpression cell lines(the miR-30-5 p group)and control cell lines(the miR-control group)in esophageal cancer cell line EC109. The EdU staining was used to analyze the proliferation in two groups. Transwell was used to analyze migration in two groups. Bioinformatics and double luciferase reporter gene were used to analyze target genes of miR-30-5 p. Influence of overexpression gene on cell proliferation and migration in the miR-30-5 p group and the miR-control group was analyzed. Results:Compared with adjacent tissues,the expression of miR-30-5 p of esophageal cancer tissues was significantly increased,with statistically significant difference(P<0.05). Compared with the miR-control group,the proliferation and migration ability in the miR-30-5 p group significantly decreased,with statistically significant difference(P <0.05). Bioinformatics and double luciferase reporter gene showed that BCL-9 was the target gene of miR-30-5 p. Overexpression of BCL-9 in the miR-control group was able to significantly increase cell proliferation and migration,with statistically significant difference(P<0.05). Over-expression of BCL-9 in the miR-30-5 p group had no statistical significance on cell proliferation and migration(P<0.05). Conclusion:The expression of miR-30-5 p is low in esophageal cancer,which can up-regulate the expression of its target gene BCL-9 and promote proliferation and migration of esophageal cancer cells.