摘要
目的探讨miR-124在膀胱癌中的表达及其临床意义。方法RT-qPCR检测人正常膀胱尿路上皮细胞SV-HUC-1及膀胱癌细胞(5637、J82、T24、TCCSUP、UM-UC-3)中miR-124的表达;收集本院2013年1月至2016年1月114例膀胱癌患者的术后组织标本,采用原位杂交法检测miR-124在膀胱癌及癌旁组织中的表达,并分析其与临床病理特征及预后的相关性。细胞试验分3组:分别将miRNA-NC(miRNA-NC组)、miRNA-124 mimics(miRNA-124 mimics组)和miR-124 inhibitor(miR-124 inhibitor组)转染至膀胱癌T24细胞,CCK-8试验检测miR-124表达对细胞增殖能力的影响,Western blot检测其增殖相关蛋白(PCNA、Ki-67)和p-AKT表达。结果qPCR结果显示,miR-124在膀胱癌细胞5637、J82、T24、TCCSUP、UM-UC-3中的表达均低于正常膀胱尿路上皮细胞SV-HUC-1(P<0.05);原位杂交试验结果显示,miR-124在正常膀胱尿路上皮细胞中呈高表达,在非浸润性和浸润性膀胱癌组织中低表达;miR-124表达与膀胱癌临床T分期、淋巴结转移及肿瘤分化程度密切相关(χ^2=6.829,P=0.009;χ^2=6.844,P=0.009;χ^2=5.111,P=0.024);Kaplan-Meier分析显示,miR-124低表达组患者的总生存率(OS)和无病生存期(DFS)明显低于高表达组患者(χ^2=22.7,P<0.01;χ^2=26.57,P<0.01)。多因素分析结果表明miR-124低表达是影响膀胱癌患者预后的独立危险因素(OS HR=4.63;95%CI:1.39~7.23;P=0.007;DFS HR=5.82;95%CI:1.56~11.42;P=0.003)。与miRNA-NC组比较,miR-124mimics组T24细胞增殖能力明显下降(P<0.05),增殖相关蛋白(PCNA、Ki-67)和p-AKT表达量明显降低(P<0.05);而miR-124 inhibitor组中T24细胞增殖能力增强,PCNA、Ki-67及p-AKT的蛋白表达量升高(P<0.05)。结论miR-124在膀胱癌细胞及组织中低表达,并促进膀胱癌细胞的增殖能力,低表达miR-124的膀胱癌患者预后较差,其机制可能与AKT信号通路的激活有关。
Objective To investigate the expression of microRNA-124(miR-124)in bladder cancer and its clinical significance.Methods The expression of miR-124 in a normal bladder urothelial cell line(SV-HUC-1)and 5 bladder cancer cell lines(5637,J82,T24,TCCSUP and UM-UC-3)was detected using RT-qPCR.We also detected the expression of miR-124 using in situ hybridization in 114 tissue specimens of bladder cancer and adenocarcinoma tissues from patients undergoing surgeries in our hospital between January,2013 and January,2016.The correlations of miR-124 expression with the clinicopathological characteristics and prognosis of the patients were analyzed.In the in vitro experiment,we tested the effect of transfection with miR-124 mimics and miR-124 inhibitor on the proliferation of bladder cancer T24 cells using CCK-8 assay and on the cellular expression of PCNA,Ki-67 and p-AKT using Western blotting.Results The expression of miR-124 in the 5 bladder cancer cell lines was significantly lower than that in normal bladder urothelial cells.The results of in situ hybridization showed that miR-124 was highly expressed in normal bladder urothelial cells but lowly expressed in non-invasive and invasive bladder cancer tissues.The expression of miR-124 was closely correlated with clinical T stage(Chi-square=6.829,P=0.009),lymph node metastasis(Chi-square=6.844,P=0.009)and tumor differentiation(Chi-square=5.111,P=0.024).Kaplan-Meier analysis showed that compared with those with a high expression of miR-124,the patients with a low expression of miR-124 had significantly shorter overall survival(Chi-square=22.7,P<0.01)and disease-free survival(Chi-square=26.57,P<0.01).Multivariate analysis showed that a low expression of miR-124 was an independent risk factor for a poor prognosis in patients with bladder cancer in terms of both the overall survival(HR=4.63,95%CI:1.39~7.23;P=0.007)and disease-free survival(HR=5.82,95%CI:1.56~11.42;P=0.003).In the cell experiment,transfection with the miR-124 mimics significantly attenuated the proliferation of T
作者
王阳
胡跃世
李征
朱清
段启新
刘磊
马慧利
李明林
WANG Yang;HU Yueshi;LI Zheng;ZHU Qing;DUAN Qixin;LIU Lei;MA Huili;LI Minglin(Department of Urology,Nanyang Central Hospital,Nanyang,Henan Province,473009,China;Department of Oncology,Nanyang Central Hospital,Nanyang,Henan Province,473009,China)
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2020年第10期1035-1041,共7页
Journal of Third Military Medical University
