miR-148a对食管癌细胞迁移及侵袭的影响及潜在分子机制研究

Role and potential mechanism of miR-148a in migration and invasion of esophageal carcinoma

摘要:

目的探究微小核糖核酸148a（miR-148a）在食管鳞状细胞癌（ESCC）中的表达及对食管癌TE-1 细胞迁移和侵袭能力的影响。方法收集48例食管癌患者肿瘤及癌旁组织的RNA标本，逆转录PCR合成互补脱氧核糖核酸（cDNA），实时荧光定量聚合酶链反应PCR检测miR-148a 在肿瘤及癌旁组织中的表达情况，分析肿瘤组织中miR-148a 表达差异的临床意义。将人工合成的miR-148a模拟物（miR-148a mimics）转染入食管癌细胞TE-1中，划痕愈合实验检测过表达miR-148a对TE-1细胞迁移的影响，Transwell小室模型检测miR-148a 对TE-1 侵袭的影响。Western blot检测miR-148a 过表达对TE-1 细胞中原癌基因（c-myc）及基质金属蛋白酶9（MMP-9）蛋白表达的影响。结果MiR-148a在食管癌组织中表达水平下调（P =0.031）；过表达miR-148a能够抑制TE-1细胞迁移（P =0.021）及侵袭（P =0.018）并下调细胞内c-myc（P =0.037）及MMP-9（ P=0.026）的表达水平。结论miR-148a 在食管癌中低表达，miR-148a可能通过抑制c-myc/MMP-9 通路的表达来发挥抑制食管癌细胞迁移及侵袭。

Abstract:

Objective To investigate the expression of miR-148a in esophageal squamous cell carcinoma (ESCC) and its role in cell migration and invasion. Methods A total of 48 cases of ESCC and tumor adjacent tissue ribonucleic acid (RNA) samples were collected in our hospital. Complementary deoxyribonucleic acid (cDNA) was synthetized by reverse transcription polymerase chain reaction (RT-PCR). The expression level of miR-148a was detected by real-time PCR. MiR-148a mimics were synthesized and transfected into TE-1 cells. Migration and invasion rates were determined by wound-healing assay and Transwell assay, respectively.The expression levels of c-myc and matrix metalloprotein 9 (MMP-9) were measured by Western blot. Results miR-148a was down-regulated in ESCC tissue (P = 0.031). Over-expression of miR-148a inhibited migration and invasion rates of TE-1 cells (P = 0.021 and 0.018, respectively). Moreover, protein levels of c-myc and MMP-9 were decreased with miR-148a over expression (P = 0.037 and 0.026, respectively). Conclusions miR-148a inhibits cell migration and invasion of ESCC through inhibition of c-myc/MMP-9 signal pathway.