过表达核糖核酸酶抑制因子对膀胱癌移植瘤侵袭转移及上皮细胞间质转化的影响

摘要

Objective To investigate the effect of over-expression of human ribonuclease inhibitor suppresses invasion and migration of transplanted bladder cancer .Methods The T24 cells were stably transfected with pIRES2-EGFP-RI and pIRES2-EG-FP plasmid respectively .Using the cell transfected with pIRES2-EGFP and untransfected cell as controls .and the positive clones were screened by G418 ,respectively ;Tumor cells of the three groups at 2 × 106 were respectively injected into the back of BALB/C nude mice to establish the xenograft models .Change of micro-blood vessels in tumor tissue and expression of CD31 were detected by Immunohisto-chemical and HE staining .Immunohisto-chemical assay was used to detect the expression of RI ,MMP-2 ,MMP-9 ,E-cadherin ,N-cadherin ,Vimentin ,Snail ,Slug ,Twist in the tumors .Results Animal experiment showed that the T24-RI cells group significantly inhibited the growth of bladder cancer compared with the other two control groups .Compared with the T24 and T24 vector cells groups ,the microvessel density in tumor tissue of T24-RI group was notably reduced and the expressions of MMP-2 , MMP-9 ,N-cadherin ,Vimentin ,Snail ,Slug ,Twist significantly were decreased simultaneously ,while the expressions of RI and E-cadherin were increased .Conclusion up-regulation RI can inhibit the growth of transplanted bladder cancer in nude mice by decrea-sing the expression of invasion protein and EM T protein .%目的：研究过表达核糖核酸抑制因子（RI）对膀胱癌移植瘤侵袭转移及上皮细胞间质转化（EM T ）的影响。方法将 RI 过表达质粒（pIRES2-EGFP-RI）和阴性对照质粒（pIRES2-EGFP）稳定转染膀胱癌 T24细胞，稳定表达细胞株分别命名为：T24-RI 组、T24 vector 组、T24组。将各组 T24细胞分别接种至 BALB／C 裸鼠左腋皮下，观察移植瘤的生长、微血管密度及肺组织转移灶的变化；进一步分析 RI 、MMP-2、MMP-9、EM T 相关蛋白（E-cadherin 、N-cadherin 、Vimentin 、Snail 、Slug 、Twist 蛋白）在肿瘤组织中的表达水平。结果 T24-RI 组较 T24组与 T24 vector 组，瘤体质量显著降低（P ＜0．01），抑瘤率分别为26．67％和38．85％；与对照组比较，T24-RI 组瘤组织微血管密度明显减少，肺组织未见明显转移灶，同时 CD31在瘤组织中表达减少；免疫组织化学染色显示：T24-RI 组瘤组织中 RI 、E-cadherin 蛋白的表达明显升高，而 MMP-2、MMP-9、N-cadherin 、Vimentin 、Snail 、Slug 、Twist 蛋白表达明显降低。结论过表达 RI 可能通过调节侵袭转移 EM T 关键蛋白的表达，显著抑制了膀胱癌移植瘤生长、侵袭和转移的潜能。