Numb基因对膀胱癌细胞侵袭和迁移能力的影响及机制研究  

Effect of Numb overexpression on invasion and migration in human bladder cancer cell line T24

作　　者：司马晋[1] 张保[1] 马鑫[2] 王保军[2] 艾青[2] 张旭[2] 

机构地区：[1]航天中心医院(北京大学航天临床医学院)泌尿外科,北京100049 [2]解放军总医院泌尿外科

出　　处：《中华泌尿外科杂志》2014年第3期226-229,共4页Chinese Journal of Urology

基　　金：国家自然科学基金（30972982）;国家自然科学基金（81100561）

摘　　要：目的 研究Numb基因对膀胱癌细胞侵袭和迁移能力的影响及相关机制.方法 2013年6月我们使用Numb-ORF表达质粒和pCMV6-Entry空载体质粒转染人膀胱癌细胞T24,作为实验组和阴性对照组,无质粒转染的人膀胱癌细胞T24作为空白对照组,采用荧光定量PCR和蛋白质印迹技术检测各组的Numb和基质金属蛋白酶-9（MMP-9）的表达情况,采用细胞划痕实验和Transwell小室法分别检测细胞的迁移和侵袭能力.结果 荧光定量PCR检测显示,与阴性对照组（△CT值5.871±0.685）和空白对照组（△CT值5.618±0.597）比较,实验组中Numb表达（△CT值1.655±0.432）显著升高（P＜0.01）.实验组的MMP-9表达（△CT值8.423±0.202）低于阴性对照组（△CT值7.294±0.138）和空白对照组（△CT值7.220±0.118）,差异有统计学意义（P＜0.01）.划痕实验中,12h的划痕修复比实验组为0.525±0.037,阴性对照组为0.693±0.034,空白对照组为0.701±0.038,组间比较差异有统计学意义（P=0.004）.侵袭实验中,实验组的穿膜细胞数为（12.6±3.2）个,明显少于阴性对照组[（130.8±9.2）个]和空白对照组[（132.2±10.6）个],差异有统计学意义（P＜0.05）.结论 上调Numb基因可以降低人膀胱癌细胞T24的侵袭和迁移能力,其机制可能是通过Numb负性调节MMP-9实现的.Objective To study the effect of Numb overexpression on invasion and migration in hu- man bladder cancer eell line T24, and its related mechanism. Methods In June 2013, the plasmids in- eluding Numb-ORF plasmid and its blank vector pCMV6-Entry were transfected into T24 cells, and set the blank control as usual. The expressions of Numb and MMP-9 （ Matrix metalloproteinase-9） were detected by Real-Time PCR and Western Blot. Then T24 cell invasion and migration were measured respectively by Tran- swell assay and Scratch Wound Healing Assay. Results Cnmpared to negative cnntrol group, which trans- leered with blank-vector plasmid and untransfected group, Numb expression level in Numb-ORF group was significantly higher （P〈0.01）. Meanwhile, MMP-9 level in Nmnb-ORF group was decreased, /~ C^r-value as h^llows： Numb-ORF group was 8.423-＋0.202, negative control group was 7.294＋0.138, untransfeeted group was 7.220.＋0.118 （P〈O.01）. In Scratch Wound Healing Assay, the repair ratio of Numb-ORF group was less than negative control group and untransfected group. The repair ratio in 12 hours as follow： Numb-ORF group was 0.525±0.037, negative control group was 0.693±0.034, untransfeeted group was 0.701 ±0.038 （P= 0.004）. In Transwell assay, the number of invasion cells in Numb-ORF group was 12.6±3.2, less than in control group （130.8±9.2） and untransfeeted group （132.2±10.6） （P〈0.05）. Conclusions Overexpression of Numb by transfection significantly decreases the capability of T24 cell invasion and migration. This suppressed effect may be due to the down-regulation of MMP-9 expression in human bladder cancer.

关 键 词：膀胱癌 Numb基因 基质金属蛋白酶-9 侵袭 迁移 

分 类 号：R737.14[医药卫生—肿瘤]