MiR-148a调控胃癌细胞侵袭迁移的作用机制研究

Mechanism of Invasion and Metastasis in Gastric Cancer Regulated by miR-148a

目的研究miR-148a在胃癌侵袭迁移中的作用,为新肿瘤标志物的研究提供理论基础。方法荧光实时定量PCR(RT-PCR)检测miR-148a在胃癌细胞系和正常胃黏膜细胞中的表达水平,HGC-27分别转染miR-148a control/miR-148a mimic和miR-148a control/miR-148a inhibitor后检测miR-148a表达水平的改变,Transwell法检测miR-148a对HGC-27细胞侵袭和迁移的影响,Western blot法检测miR-148a对胃癌细胞中c-myc蛋白表达的调控。结果 MiR-148a在胃癌细胞中的表达明显低于正常胃黏膜细胞;HGC-27转染miR-148a mimic后miR-148a表达升高,HGC-27转染miR-148a inhibitor后miR-148a表达下降;miR-148a能够显著抑制HGC-27细胞的侵袭和迁移;miR-148a能够明显下调c-myc蛋白的表达。结论 miR-148a通过c-myc来发挥抑制胃癌侵袭迁移的作用。

Objective To study the role of miR - 148a in regulation of invasion and metastasis in gastric cancer, to provide a theoretical basis for the study of novel tumor markers. Methods In this study, the mRNA expression levels of miR - 148a were examined in GC cell lines and normal gastric mucosa cells line by quantitative reverse transcription - PCR. The functions of miR - 148a in migration/ invasion were determined by using transwell. The expression of miR - 148a in the HGC -27 cell after tranfected with miR - 148a mimic or miR - 148a inhibitor was detected using qRT - PCR. The protein expression of c - myc was measured by western blot. Results The ex- pression of miR - 148a was downregulated in the GC cell lines （ SGC - 7901, MGC - 803, HGC - 27, MKN - 45 and BGC - 823 ） com- pared with normal gastric mucosa cells line （ GES - 1 ）. After tranfected with miR - 148a mimic, the expression of miR - 148a increased in HGC - 27 cell, but miR - 148a inhibitor decreased the expression of miR - 148a in HGC - 27. HGC - 27 infected with miR - 148a inhibi- tots showed increased cell invasion and migration. We also observed that HGC - 27 cell expressing miR - 148a mimics exhibited reduced migration rates compared with the controls. Our results show that ectopic expression of miR - 148a decreased c - myc in gastric cancer cell line. Conclusion miR - 148a suppresses gastric cancer cell invasion and migration by inhibiting c - myc.