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

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(c DNA),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 Mi R-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。

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 adjacenttissue 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 ofmiR-148a was detected by real-time PCR. MiR-148a mimics were synthesized and transfected into TE-1cells. Migration and invasion rates were determined by wound-healing assay and Transwell assay, respectively.The expression levels of c-myc and matrix metalloprotein9 （MMP-9） were measured by Western blot. ResultsmiR-148a was down-regulated in ESCC tissue （ = 0.031）. Over-expression of miR-148a inhibited migrationand invasion rates of TE-1 cells （ = 0.021 and 0.018, respectively）. Moreover, protein levels of c-myc andMMP-9 were decreased with miR-148a over expression（ = 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.