摘要
目的探讨微小RNA-204(microRNA-204,miR-204)在食管鳞状细胞癌(esophageal squamous cell carcinoma,ESCC)组织中的表达、临床意义及可能分子机制。方法收集2014-01-04-2014-11-25滨州医学院附属医院手术切除的60例ESCC及对应癌旁组织。实时定量PCR(quantitative real-time PCR,qRT-PCR)检测miR-204、Bcl-2及SIRT1mRNA在ESCC及癌旁组织中的表达,统计分析miR-204表达与肿瘤临床病理特征及Bcl-2和SIRT1mRNA的相关性。采用化学合成的miR-204模拟物转染人食管癌TE-1细胞,MTT法和流式细胞术检测TE-1细胞的增殖和凋亡情况,qRT-PCR和蛋白质印迹法分别检测miR-204下游潜在靶点Bcl-2与SIRT1的mRNA和蛋白表达。结果 miR-204在ESCC组织中表达水平显著低于对应癌旁组织,t=4.752,P<0.001;ESCC组织中miR-204低表达与肿瘤大小(t=2.847,P=0.006)及肿瘤TNM分期(t=3.341,P=0.002)显著相关;ESCC组织中miR-204与Bcl-2和SIRT1的mRNA表达呈负相关关系,P<0.002;miR-204可显著抑制TE-1细胞的增殖并促进其凋亡,P<0.05;上调miR-204表达可显著抑制Bcl-2与SIRT1的mRNA与蛋白表达水平,P<0.05。结论 miR-204在ESCC组织中表达下调,并与肿瘤恶性临床病理特征有关。miR-204可能通过下调Bcl-2和SIRT1的表达来发挥抑制食管癌细胞生长的生物学作用。
OBJECTIVE To investigate the expression level of microRNA-204 （miR-204） in human esophageal squamous cell carcinoma （ESCC）, its potential role in ESCC tumorgenesis and realated molecular mechanism. METttODS Quantitative re- al-time PCR （qRT-PCR） was applied to detect the expression of miR-204 in primary ESCC （n= 60） and matched tumor adjacent tissues. The relationship between the miR 204 expression and clinical features and mRNA level of Bcl-2 or SIRT1 were analyzed by Pearson chi-square test. miR-204 mimics were transiently transfected into human TE-1 cells in vitro. The proliferation and apoptosis of cells were detected by MTT assay and flow cytometry, mRNA and protein levels of Bcl-2 and SIRT1 were measured by qRT-PCR and western blot, respectively. RESULTS The expression level of miR-204 in ESCC tissues was significantly low- er than that in tumor adjacent tissues （t=4. 752, P〈0. 001）, which was also associated with tumor size （t=2. 847,P=0. 006） and advanced TNM stage （t= 3. 341, P= 0. 002）. There was both negative correlation between the expression of miR-204 and Bcl-2 mRNA and between miR-204 and SIRT1 mRNA （P〈0.00l）. Overexpression miR-204 in TE-1 cells suppressed prolifera tion and enhanced apoptosis （P%0. 05） as well as down-regulated mRNA and protein expressions of Bcl-2 and SIRT1 （P〈 0.05）. CONCLUSIONS The expression of miR-204 in ESCC tissues was significantly lower than in tumor adjacent tissues. miR-204 could inhibit ESCC cell proliferation and induce apoptosis by down-regulating the expression of Bcl-2 and SIRT1.
出处
《中华肿瘤防治杂志》
CAS
北大核心
2015年第17期1382-1387,共6页
Chinese Journal of Cancer Prevention and Treatment
