KAI1蛋白促进786-O细胞黏附及侵袭作用的研究

Research of KAI1 on the adhesion and invasion of human renal cell carcinoma cell line 786-O

目的观察KAI1蛋白对肾透明细胞癌786-O细胞黏附及侵袭的影响,初步研究KAI1对786-O细胞黏附、侵袭作用机制。方法逆转录PCR、蛋白免疫印迹、免疫细胞化学鉴定pCMV-KAI1细胞KAI1表达情况;细胞黏附实验及迁移实验检测细胞黏附及迁移能力;蛋白免疫印迹检测细胞黏附因子CD44及肿瘤细胞迁移抑制因子MRP-1/CD9的表达。结果与空白对照和阴性对照组相比,pCMV-KAI1细胞中KAI1的mRNA转录水平显著升高(P<0.01);pCMV-KAI1细胞的黏附能力及侵袭能力都显著低于其他两组(P<0.01);免疫细胞化学结果证实KAI1阳性染色细胞数目接近100%,且显著地高于其他两组;蛋白免疫印迹实验结果证实,pCMV-KAI1细胞中CD44的表达显著降低,而MRP-1/CD9的表达显著升高。结论 KAI1通过抑制黏附因子CD44的表达降低了细胞的黏附性,通过促发肿瘤转移抑制因子MRP-1/CD9表达的升高而降低其侵袭性。

[Objective] To observe the effect of KAI1 protein on adhesion and invasion of renal carcinoma cell 786-0 and study the mechanisms of KAI1 protein on adhesion and invasion of renal carcinoma cell 786-0. [Methods] Using the retro-transcription PCR and Immunocytochemistry methods to detect the expression of KAI1 in the pCMV- KAI1 cell. The invasive and adhesive ability of pCMV- KAI1 cell were studied by the migration assay and Boyden Chamber assay, respectively. Cell adhesive factor Cd44 and MRP-1/CD9 were detected by the western blot assay. [Results] The transcription level of KAI1 mRNA was significantly increased in the pCMV-KAI1 cell compared with the pCMV-NEG and CON cell （P〈0.01）; pCMV- KAI1 cell adhesion and invasion were significantly lower than the other two groups （P 〈0.01）; Immunocytochemistry confirmed that the number of positive staining KAI1 cells close to 100%, and significantly higher than the other two groups; The CD44 protein was decreased significantly and MRP-1/CD9 was increased significantly, respectively, compared with the other groups （P 〈0.01）. [Conclusion] KAI1 inhibited the adhesive ability of pCMV-KAI1 cell by blocking the CD44 expression, and inhibited the invasive ability by up-regulating the MRP-1/CD9 expression.