Effect of Numb overexpression on invasion and migration in human bladder cancer cell line T24 / 中华泌尿外科杂志

ABSTRACT

Objective To study the effect of Numb overexpression on invasion and migration in human bladder cancer cell line T24,and its related mechanism.Methods In June 2013,the plasmids including 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 Transwell assay and Scratch Wound Healing Assay.Results Compared to negative control group,which transfected 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 Numb-ORF group was decreased,△ CT-value as followsNumb-ORF group was 8.423±0.202,negative control group was 7.294±0.138,untransfected group was 7.220±0.118 (P＜0.01).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 followNumb-ORF group was 0.525±0.037,negative control group was 0.693±0.034,untransfected 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 untransfected 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.