摘要
目的研究人膀胱尿路上皮癌细胞和组织中的mi R-128与血管内皮生长因子C(VEGF-C)的靶向调控关系及其生物学功能。方法收集9对配对的膀胱癌组织和其癌旁组织,1株人正常尿路上皮细胞(SV-HUC-1)和4株膀胱癌细胞(T24、5637、UM-UC-3和RT4),通过RT-PCR、Western Blot方法检测VEGF-C和mi R-128的表达水平;将mi R-128复合物或mi R-128抑制剂等复合物转染到2株膀胱癌T24和5637细胞中,检测mi R-128对其生长、迁移及侵袭的影响;生物信息学软件预测分析mi R-128的靶基因,双荧光素酶实验验证mi R-128是否靶向作用VEGF-C。结果相比癌旁组织和正常尿路上皮细胞,膀胱癌组织和细胞中mi R-128的表达明显下调(P<0.05),而VEGF-C在癌组织和细胞中都呈现高表达(P<0.05)。mi R-128可抑制膀胱癌细胞生长、克隆形成及迁移侵袭(P<0.05)。双萤光素酶报告实验证实mi R-128靶向作用VEGF-C(P<0.05)。结论 mi R-128在膀胱癌中表达下调,而VEGF-C表达上调;mi R-128可通过靶向调控VEGF-C影响膀胱癌细胞的增殖和转移。
【Objective】To investigate the regulation of mi R-128 on VEGF-C expression and its biological effect in bladder cancer. 【Methods】Materials including 9 pairs of bladder cancer tissues and adjacent tissues, 4 bladder caner cell lines（T24, 5637, UM-UC-3, RT4） and 1 normal urothelial cell（SV-HUC-1） were collected. The mi R-128 and VEGF-C m RNA and protein expression were tested by RT-PCR and Western blot, respectively. The mi R-128 minics and mi R-128 inhibitor were transfected into bladder cancer cells T24 and 5637 through lipidosome. And the effect of mi R-128 on bladder urothelial cancer cells proliferation, invasion and metastasis were tested. Target gene of mi R-128 analysis was applied by related biological information software. Dual luciferase reporter gene assay verified targeted relationship between mi R-128 and VEGF-C.【Results】We found mi R-128 was down-regulated in bladder cancer tissues and cell lines which was opposite from the expression of VEGF-C（P〈0.05）. Overexpression of mi R-128 could inhibit proliferation, migration and invasion of bladder cancer cells（P〈0.05）. Dual luciferase reporter gene assay showed VEGF-C was a direct target of mi R-128 in bladder cancer cells（P〈0.05）.【Conclusions】mi R-128 was down-regulated in bladder cancer, meanwhile VEGF-C was up-regulated. Mi R-128 down-regulation can promote the growth and metastasis of bladder cancer cells by involving VEGF-C up-regulation.
出处
《中国现代医学杂志》
CAS
北大核心
2015年第25期38-44,共7页
China Journal of Modern Medicine
作者简介
[通信作者]周旭，E-mail：zx315zx@163．com
