MAP4K4在膀胱癌中的表达及对膀胱癌细胞株T24生物学行为的影响  

Expression of MAP4K4 in bladder cancer and its biological effect on human bladder cancer T24 cells

作　　者：马宝杰[1] 李海波[1] 张昌文[1] 蔡启亮[1] 李刚[1] 徐勇[1] 

机构地区：[1]天津医科大学第二医院泌尿外科天津市泌尿外科研究所,300211

出　　处：《中华泌尿外科杂志》2014年第6期469-472,共4页Chinese Journal of Urology

基　　金：天津市高等学校科技发展基金计划项目（20120133）

摘　　要：目的 探讨丝裂原激活蛋白激酶激酶激酶激酶4（MAP4K4）对膀胱癌细胞株T24增殖、侵袭和凋亡的影响. 方法 采用基因芯片技术对2010年6月至201 1年6月5对肌层浸润性膀胱癌及癌旁组织的基因表达谱进行初步研究,对比癌和癌旁组织的差异基因表达,筛选与膀胱癌发生、发展相关基因.结合文献检索,筛选出 MAP4K4作为研究基因,并采用PCR和蛋白质印迹法从转录和翻译水平检测16对肌层浸润性膀胱癌及癌旁组织中MAP4K4的表达情况,验证基因芯片的结果.然后设计一个空载体质粒（对照组）及MAP4K4靶向短发夹RNA （shRNA）核苷酸序列质粒（靶向干扰组）分别转染T24细胞,嘌呤霉素筛选稳定转染细胞,qRT-PCR和蛋白质印迹法检测靶基因干扰效率,并对转染后T24细胞进行MTT细胞增殖实验、Transwell细胞侵袭实验及Annexin-V法细胞凋亡检测. 结果 膀胱癌组织中MAP4K4表达高于癌旁组织（P=0.020）.MAP4K4基因沉默后细胞增殖减慢（靶向干扰组与对照组,P＜0.001）.靶向干扰组与对照组穿膜细胞分别为5.40±2.07和17.00±2.74（P=0.004）,细胞凋亡比例分别为（15.72±6.40）％和（4.23±3.29）％（P=0.023）,组间比较差异均有统计学意义. 结论 MAP4K4在膀胱癌中相对于正常组织高表达,降低MAP4K4基因表达可以抑制膀胱癌细胞的增殖、侵袭,促进细胞凋亡.MAP4K4有望成为膀胱癌治疗的新靶点.Objective To observe the effect of MAP4K4 targeted shRNA on biological characteristics such as proliferation,invasiveness,and apoptosis in human bladder cancer cell.Methods Differentially expressed genes was screened out through cDNA microarray analysis in 5 pairs of fresh-frozen muscle-invasive bladder cancer（MIBC） and adjacent normal tissue obtained from radical cystectomy.Combining the results of genechip and literature review,MAP4K4 was picked up for further analysis.To verify the result of microarray analysis,16 pairs of fresh muscle-invasive bladder cancer （MIBC） and adjacent tissues were assessed for the expression of MAP4K4 mRNA and protein through RT-PCR,qRT-PCR and Western-blot.T24 cell line was stably trasfected with MAP4K4 targeted shRNA and control shRNA,respectively.The effects of MAP4K4 silencing on proliferation,invasiveness and apoptosis of T24 cells transfected with MAP4K4 targeted shRNA and control shRNA were assessed by 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide （MTT）,transwell and flowcytometry （FCM） assay.Results MAP4K4 was overexpressed in muscle invasive bladder cancer than in normal tissue.Down regulation of MAP4K4 expression decreased bladder cancer cell proliferation（MAP4K4-targeted versus control,P〈0.001）,invasiveness（MAP4K4-targeted versus control,P=0.004）and promoted cell apoptosis（MAP4K4-targeted versus control,P=0.023）.Conclusions MAP4K4 is overexpressed in muscle invasive bladder cancer than in normal tissue.Down-regulation of MAP4K4 expression inhibits the invasive ability of bladder cancer.Therefore,MAP4K4 might be a potential therapeutic target for bladder cancer.

关 键 词：丝裂原激活蛋白激酶激酶激酶激酶4 膀胱肿瘤 基因沉默 寡核苷酸序列分析 MITOGEN-ACTIVATED protein KINASE KINASE KINASE KINASE 4 

分 类 号：R737.14[医药卫生—肿瘤]