论文部分内容阅读
目的分析microRNA-214在膀胱癌中表达的生物学功能、调控机制以及临床应用探讨。方法选取我院2014-2015年收治的膀胱癌120例患者作为研究对象,设为实验组,取120例患者的膀胱癌新鲜冰冻组织,其中包括40例NMIBC,80例MIBC,选取120例正常人作为对照组。采用qRT-PCR方法对两组患者中的生化上皮细胞以及两种膀胱癌细胞系中的表达情况,采用体外用脂体2000将miR-214模拟物进行转染至膀胱癌细胞中,设计阴性对照组和空白对照组。采用qRT-PCR方法评估各组转染效率。利用综合生物学信息分析平台,联合五种计算方法,寻找miR-214的靶基因,并且采用双荧光素酶报告系统进行验证。采用siRNA对膀胱癌细胞系中的靶基因的表达水平进行干扰。结果 miR-214在两种膀胱癌细胞中的表达与生化上皮细胞比较明显下降,P<0.05。对患者进行随访发现,120例患者中有66例膀胱癌复发,复发率为55%。总生存患者72例,总生存率为60%。结论 miR-214分析在膀胱癌组织中出现表达水平下降,与膀胱癌负面性临床病例特征有密切联系,提示膀胱癌疾病的发生和发展,在其过程中发挥重要的作用。MiR-214在膀胱癌疾病中发挥抑制癌基因的作用,抑制膀胱癌细胞的增值和迁移,促进癌细胞凋亡。
Objective To analyze the biological function, regulatory mechanism and clinical application of microRNA-214 in bladder cancer. Methods A total of 120 patients with bladder cancer admitted to our hospital from 2014 to 2015 were enrolled as the experimental group. Fresh frozen tissues of 120 patients with bladder cancer were included, including 40 cases of NMIBC and 80 cases of MIBC. 120 normal subjects As a control group. MiR-214 mimics were transfected into bladder cancer cells using Lipofectamine 2000 in vitro using qRT-PCR method for biochemical epithelial cells in both groups and in two bladder cancer cell lines. Negative control Group and blank control group. The transfection efficiency of each group was evaluated by qRT-PCR method. Using the comprehensive biological information analysis platform, we combined five computational methods to find the target gene of miR-214 and verified it with the dual luciferase reporter system. SiRNA is used to interfere with the expression level of target genes in bladder cancer cell lines. Results The expression of miR-214 in two kinds of bladder cancer cells was significantly lower than that in biochemical epithelial cells (P <0.05). Follow-up of patients revealed that 66 of 120 patients had a recurrence of bladder cancer with a recurrence rate of 55%. 72 patients with total survival, the overall survival rate was 60%. Conclusion The decreased expression of miR-214 in bladder cancer tissues is closely related to the negative clinical features of bladder cancer, suggesting that the occurrence and development of bladder cancer diseases play an important role in the process. MiR-214 plays a role of oncogene inhibition in bladder cancer, inhibits proliferation and migration of bladder cancer cells, and promotes apoptosis of cancer cells.