下调lncRNA CTC-425F1.4靶向调控miR-146a-3p表达对膀胱癌细胞的增殖和侵袭的影响

Effect of Down-regulating lncRNA CTC-425F1.4 on the Proliferation and Invasion of Bladder Cancer Cells through Targeted Regulation of MiR-146a-3p Expression

目的研究长链非编码RNA(long-chain non-coding RNA,lncRNA)CTC-425F1.4在膀胱癌组织中的表达,探讨下调CTC-425F1.4对膀胱癌细胞增殖和侵袭的影响及相关机制。方法荧光实时定量PCR测定30例膀胱癌组织及癌旁组织、4种膀胱癌细胞株(UM-UC-3、T24、RT4、5637)和正常膀胱上皮细胞SV-HUC-1中CTC-425F1.4表达变化。在RT4细胞中分别转染CTC-425F1.4抑制剂(inhibitor组)和阴性对照序列(control组),荧光实时定量PCR检测CTC-425F1.4下调效果。CCK-8法和Transwell小室法分别检测CTC-425F1.4 inhibitor对膀胱癌细胞增殖和侵袭的影响。采用生物信息学软件预测CTC-425F1.4的靶基因,采用荧光素酶报告系统验证两者的靶向关系。荧光实时定量PCR检测CTC-425F1.4 inhibitor对膀胱癌细胞miR-146a-3p表达的影响。Western blot法检测p21、Zeb1、Vimentin、Snail蛋白的表达。结果膀胱癌组织中CTC-425F1.4表达水平高于癌旁组织(P<0.01)。膀胱癌细胞中CTC-425F1.4表达水平高于正常膀胱上皮细胞(P<0.05)。与control组比较,转染CTC-425F1.4 inhibitor的RT4细胞中CTC-425F1.4表达水平显著降低(P<0.01)。与control组比较,下调CTC-425F1.4表达抑制了RT4细胞的增殖(P<0.05)和侵袭(P<0.01)。CTC-425F1.4能够靶向调控miR-146a-3p表达。与control组比较,转染CTC-425F1.4 inhibitor的RT4细胞中miR-146a-3p表达水平显著升高(P<0.01),p21蛋白含量显著增加,Zeb1、Vimentin和Snail蛋白含量显著减少。结论CTC-425F1.4在膀胱癌组织和细胞株中高表达,下调CTC-425F1.4靶向调控miR-146a-3p的表达抑制膀胱癌RT4细胞的增殖和侵袭能力。

Objective To study the expression of long-chain non-coding RNA(lncRNA)CTC-425F1.4 in bladder cancer tissues,and to explore the effect of down-regulation of CTC-425F1.4 on the proliferation and invasion of bladder cancer cells and related mechanism.Methods Fluorescence real-time quantitative PCR was used to determine CTC-425F1.4 expression changes in 30 cases of bladder cancer tissues and adjacent tissues,4 bladder cancer cell lines(UM-UC-3,T24,RT4,5637)and normal bladder epithelial cells SV-HUC-1.CTC-425F1.4 inhibitor(inhibitor group)and negative control sequence(control group)were transfected into RT4 cells,and the down-regulation effect of CTC-425F1.4 was detected by fluorescence real-time quantitative PCR.CCK-8 method and Transwell chamber method were used to detect the effects of CTC-425F1.4 inhibitor on the proliferation and invasion of bladder cancer cells.Bioinformatics software was used to predict the target gene of CTC-425F1.4,and the luciferase reporter system was used to verify the targeting relationship.Fluorescence real-time quantitative PCR was used to detect the effect of CTC-425F1.4 inhibitor on the expression of miR-146a-3p in bladder cancer cells.Western blot method was used to detect the expression of p21,Zeb1,Vimentin and Snail protein.Results The expression level of CTC-425F1.4 in bladder cancer tissue was higher than that in adjacent tissues(P<0.01).The expression level of CTC-425F1.4 in bladder cancer cells was higher than that in normal bladder epithelial cells(P<0.05).Compared with the control group,the expression level of CTC-425F1.4 in RT4 cells transfected with CTC-425F1.4 inhibitor was significantly reduced(P<0.01).Compared with the control group,down-regulating the expression of CTC-425F1.4 inhibited the proliferation(P<0.05)and invasion(P<0.01)of RT4 cells.CTC-425F1.4 can target the expression of miR-146a-3p.Compared with the control group,the expression level of miR-146a-3p in RT4 cells transfected with CTC-425F1.4 inhibitor was significantly in-creased(P<0.01),the protein content of p21 was significantly increased,and the protein content of Zeb1,Vimentin and Snail was sig-nificantly reduced.Conclusion CTC-425F1.4 is highly expressed in bladder cancer tissues and cell lines.Down-regulation of CTC-425F1.4 inhibits the proliferation and invasion of bladder cancer RT4 cells by targeting the expression of miR-146a-3p.