MIIP对肾癌细胞增殖、侵袭及迁移能力的影响

A study of MIIP impact on the proliferation,invasion and migration in renal carcinoma cell

目的:利用质粒转染技术制备过表达迁移侵袭抑制蛋白(migration and invasion inhibitory protein,MIIP)的肾癌786-0/MIIP细胞系,观察MIIP对细胞增殖、侵袭及迁移能力的影响。方法:构建携带MIIP基因的质粒,通过脂质体质粒转染技术上调肾癌786-0细胞系MIIP基因表达。Western-blot、Real-time PCR检测转染效果;利用MTT实验检测MIIP对786-0细胞增殖能力的影响;Transwell实验、划痕实验检测MIIP对786-0细胞侵袭、迁移能力的影响。结果:Western-blot、Real-time PCR检测发现实验组786-0/MIIP细胞MIIP蛋白及mRNA表达量有效上调。MTT实验显示实验组786-0/MIIP细胞增殖能力明显减弱(P<0.01)。Transwell实验显示实验组786-0/MIIP细胞侵袭能力减弱,穿透小室细胞数明显减少(P<0.01)。细胞划痕实验显示实验组786-0/MIIP细胞迁移能力显著下降(P<0.01)。结论:通过质粒转染技术能够制备过表达MIIP的肾癌细胞系。转染成功的肾癌786-0/MIIP细胞系MIIP表达量增加,有效抑制了肾癌细胞的增殖、侵袭与迁移。

Objective ：To prepare renal carcinoma cell line 786 -0/MIIP via plasmid transfection technique, then investigate the change of renal carcinoma cellg proliferation, invasion and migration ability. Methods ：The renal carci- noma cell strain 786 -0 was transfeeted by MIIP gene plasmid. The transfeetion effects were detected by Real -time PCR and Western- blot technique. The change of eellg proliferation, invasion and metastasia ability was detected by MTT assay,Transwell assay and ground -healing assay respectively. Results： By the Western -bolt and Real -time PCR technique, the expression levels of MIIP protein and mRNA in 786 -0/MIIP cell strain were up - regulated effectively. Meanwhile, the proliferation and invasion ability of 786 - 0/MIIP group decreased significantly（ P 〈 0.01 ） , and the number of 786 -0/MIIP cell that had through the membrane was the lowest in the test（P 〈 0.01 ）. Similarly, the migration distance of 786 - 0/MIIP cell was the shortest in Wound - healing assay-（P 〈 0.01 ）. Conclusion： Through the plasmid transfection technique,the RCC cell line that could over expressed MIIP were prepared successfully and it can suppres the renal cancer eellg proliferation,invasion and metastasis.