Bmi-1 shRNA抑制膀胱癌EJ细胞侵袭和转移实验研究.docVIP

Bmi-1 shRNA抑制膀胱癌EJ细胞侵袭和转移的实验研究 　　【摘要】目的：研究Bmi-1基因沉默对膀胱癌EJ细胞侵袭和转移的影响。方法：用细胞划痕实验和Transwell侵袭实验观察Bmi-1 shRNA对膀胱癌EJ细胞侵袭和转移的影响。用RT-PCR方法检测Bmi-1 shRNA对MMP-2 mRNA表达的影响。用Westem Blotting方法检测Bmi-1 shRNA对E-Cadherin和α-SMA表达的影响。结果：Transwell侵袭实验结果显示实验组细胞的穿膜细胞数为（77.0 ±4.8）个，未转染组细胞的穿膜细胞数为（152.1 ±4.3）个，空质粒组细胞的穿膜细胞数为（155.0 ±6.5）个。细胞划痕实验结果显示实验组细胞迁移能力明显降低。RT-PCR结果显示，与不加TGF-β组相比，TGF-β作用后，未转染组和空质粒组中MMP-2表达明显升高，而Bmi-1 shRNA组MMP-2表达没有变化。Western结果显示，与不加TGF-β组相比，TGF-β作用后，未转染组和空质粒组中E-Cadherin表达明显降低和α-SMA表达明显升高，而Bmi-1 shRNA组E-Cadherin和α-SMA表达没有变化。结论：Bmi-1 shRNA可以抑制膀胱癌EJ细胞侵袭和转移，其机制可能是通过阻断TGF-β促进肿瘤细胞侵袭和转移作用完成的。 　　【关键词】Bmi-1；EJ细胞；侵袭；转移 　　【中图分类号】R737.14【文献标识码】A【文章编号】1008-6455（2011）04-0015-02 　　shRNA-silenced Bmi-1 gene inhibits the metastasis and invasiveness ofbladder carcinoma EJ cells 　　Li Chengwen1 Zhu Mingyang1 Li IHui1 Chang Jiwu2 　　【Abstract】Objective:Blocking the expression of Bmi-1 with shRNA interference techniques，the effects of suppressing the metastasis and invasiveness of human bladder carcinoma EJ cells were investigated.Methods:The ability of migration and invasion of EJ cells were observed by Transwell and Wound closure assay. The expression of MMP-2, E-Cadherin and α-SMA were analyzed by RT-PCR and Western blotting. Results:In invasion experiment, the trans-membrane cell number in pGeneClipTM-Bmi-1 transfected cells was 77.0±4.8, and that in untransfected cells and in blank plasmid transfected cells was 152.1±4.3 and 155.0±6.5 respectively. The migration were inhibited significantly in EJ cells transfected by pGeneClipTM-Bmi-1 compared with that in EJ cells transfected by blank plasmid and untransfected cells. The expression change of MMP-2, E-Cadherin and α-SMA in presence of TGF-β were inhibited in pGeneClipTM-Bmi-1 transfected cells. Conclusion Bmi-1 shRNA suppresses the migration and invasiveness of EJ cells and perform the function through blocking the TGF-β signaling pathway. 　　【Key words】Bmi-1; EJ cells; invasiveness；metastasis 　　原癌基因Bmi-1属于多疏基基因家族，在细胞生长、增殖和干细胞自我更