摘要
目的:探讨miR-34a调控生存素(Survivin)表达对人膀胱移行细胞癌T24细胞生物学行为的影响。方法:以实时定量逆转录PCR技术(qRT-PCR)和Western Blot免疫印迹技术分别检测人膀胱移行细胞癌T24细胞在转染miR-34a模拟物前后miR-34a和Survivin蛋白的相对表达量。同时以四甲基偶氮唑盐比色法(methyl-thiazol-tetrazolium,MTT)实验、划痕实验和Transwell实验分析转染后T24细胞生长、迁移及侵袭能力的改变。结果:与正常人膀胱上皮细胞SV-HUC-1细胞相比,T24细胞miR-34a丰度明显降低而Survivin蛋白表达明显上调(P<0.05)。在稳定转染miR-34a模拟物后,随着T24细胞中miR-34a表达丰度上升,Survivin蛋白的表达明显受抑(P<0.05)。而转染miR-34a模拟物后的T24细胞与转染前相比,其细胞增殖率、迁移和侵袭能力也显著降低(P<0.05)。结论:miRNA-34a可以通过抑制其靶基因Survivin影响人膀胱移行细胞癌T24细胞的多种生物学,是膀胱癌潜在的治疗靶点。
Objective： To explore the regulation of Survivin protein by miR-34a and its effect on biological behavior of human bladder cancer cells.Methods： The relative expression levels of miR-34a and Survivin protein were quantitated by qRT-PCR and Western Blot in human bladder cancer cell line T24 before and after tansfect miR-34a mimics.Finally,proliferation,migaraion and invasion of T24 cells were examed in vivo by MTT,wound healing assay and transwell assay.Results： Relative quantification showed lower expression level of miR-34a and higher level of Survivin protein in T24 cell line than in human fetal bladder cell line SV-HUC-1（P0.05）.However,down-regulation of Survivin by miR-34a was found after transfected miR-34a mimics in T24 cells,and that significantly inhibits the cells proliferation,migaraion and invasion in vivo（P0.05）.Conclusion： Down-regulation of Survivin protein by miR-34a would inhibit cells proliferation,invasion and metastasis in human bladder cancer cells.miR-34a was probably one of potential molecules for the treatment of human bladder cancer.
出处
《南通大学学报（医学版）》
2012年第5期381-385,F0003,共6页
Journal of Nantong University(Medical sciences)
