微小RNA-1-3p的表达水平对胃癌细胞增殖侵袭的影响

Expression of microRNA-1-3p in gastric cancer tissues and its effect on proliferation and invasion of gastric cancer cells

目的观察微小RNA(miR)-1-3p在胃癌(GC)细胞和组织中的表达,探讨miR-1-3p对GC细胞增殖侵袭能力的影响及可能的分子调控机制。方法收集2014年1月至2015年6月在兰州大学第一医院行手术治疗的82例GC患者组织标本和癌旁组织。采用实时荧光定量聚合酶链反应(RT-qPCR)检测miR-1-3p在GC组织及癌旁组织、人GC细胞系(MKN-45、BGC823、AGS、MGC803、SGC7901)和人胃黏膜上皮细胞(GES-1)中的表达水平。分析miR-1-3p表达水平与GC患者临床病理特征参数及预后的相关性,用克隆形成实验和Transwell侵袭实验分别检测miR-1-3p的表达水平对GC细胞增殖侵袭能力的影响。采用生物信息学和荧光素酶报告基因实验验证miR-1-3p与B细胞淋巴瘤/白血病-2相关永生基因4(BAG4)的靶向调控关系。结果miR-1-3p在GC组织中的相对表达量(0.815±0.060)低于癌旁组织(1.604±0.140,t=5.922,P<0.01),其表达水平与肿瘤大小、浸润深度、淋巴结转移和TNM分期密切相关(均P<0.05)。miR-1-3p低表达组患者的总生存率低于高表达组[风险比(HR)=0.519,95%可信区间(CI):0.259~0.953,P<0.05)。过表达miR-1-3p组GC细胞的增殖和侵袭能力显著低于对照组(t=9.089,P<0.01),敲低miR-1-3p后得到了与之相反的结果(t=8.783,P<0.01)。BAG4是miR-1-3p潜在的靶基因。下调miR-1-3p可逆转si-BAG4对GC细胞增殖和侵袭能力的作用。结论miR-1-3p通过靶向BAG4的表达调控GC细胞的增殖侵袭。

Objective To investigate the expression of microRNA(miR)-1-3p in gastric cancer(GC)cells and tissue,and ectopic expression of miR-1-3p in regulating proliferation and invasion ability,and to explore its possible molecular mechanism.Methods The expression level of miR-1-3p in GC tissues and paracancerous tissues as well as human GC cell lines(MKN-45,BGC823,AGS,MGC803,SGC7901)and human gastric mucosal epithelial cells(GES-1)were detected by quantitative real-time polymerase chain reaction(RT-qPCR).The correlation between miR-1-3p expression level and clinicopathological characteristic parameters and prognosis of GC patients was analyzed.The effects of miR-1-3p expression level on the proliferation and invasion ability of GC cells were detected by clone formation assay and Transwell invasion assay,respectively.Bioinformatics and luciferase reporter gene assay were used to verify the targeted regulatory relationship between miR-1-3p and B cell lymphoma/leukemia-2 associated athanogene 4(BAG4).Results The expression of miR-1-3p in GC tissues(0.815±0.060)was significantly lower than that in paracancerous tissues(1.604±0.140,t=5.922,P<0.01).The expression of miR-1-3p was correlated with tumor size,tumor invasion depth,lymph node metastasis and TNM stage in GC patients(all P<0.05).The overall survival rate of GC patients with low expression of miR-1-3p was significantly lower than that of patients with high expression of miR-1-3p[hazard ratio(HR)=0.519,95%confidence interval(CI):0.259-0.953,P<0.05].The proliferation and invasion ability of GC cells was significantly decreased after miR-1-3p overexpression(t=9.089,P<0.01).Knockdown of miR-1-3p achieved the opposite results(t=8.783,P<0.01).Downregulation of miR-1-3p could reverse the effects of si-BAG4 on proliferation and invasion of GC cells.Conclusion MiR-1-3p regulates the proliferation and invasion of GC cells by targeting the expression of BAG4.