摘要
目的:探讨体外沉默葡萄糖调节蛋白GRP78/BIP表达对膀胱癌T24细胞侵袭及迁移的影响及可能机制。方法:设计、合成靶向GRP78基因的小干扰RNA,利用脂质体Liopfecta mineRNAIMAX转染入膀胱癌T24细胞,分别采用Realtime PCR、Western blot在mRNA和蛋白水平检测细胞内GRP78、MMP2、MMP9、Timp-2表达,采用Transwell实验及细胞划痕实验检测沉默GRP78表达对膀胱癌T24细胞侵袭及迁移的影响。结果:实验(siRNA-GRP78)T24细胞中GRP78表达显著降低,细胞侵袭迁移能力明显受到抑制,MMP2、MMP9表达降低,而Timp-2表达增高,差异均有统计学意义(P<0.05)。结论:沉默GRP78能明显抑制人膀胱癌细胞的侵袭、迁移能力,其机制可能与影响MMP2、MMP9、Timp-2表达相关。
Objective： To investigate effect of silencing GRP78 in the invasion and metastasis of Bladder cancer T24 cells and the possible mechanism. Methods： The siRNA targeting GRP78 gene was chemically synthesized,and transfect into human bladder cancer T24 cells by Liopfecta mineRNAIMAX. The mRNA and protein expression levels of GRP78,MMP2,MMP9 and Timp-2 were detected by Real-time PCR and Western blot. Transwell assay and Scratch Wound Assay were conducted to deter mine the potency of migration and invasion of T24 cells. Results： The expression of AEG1 mRNA and protein were significantly down-regulated in T24 cells transfected with siRNA targeting GRP78 as compared with the control group（ P〈 0. 01）; so was expression levels of MMP2 and MMP9protein（ P 〈0. 05）; but timp-2 expression was up-regulated. Cell migration and invasion capacity of T24 cells tranfected with GRP78 siRNA group had lower than the cells in the transfection with siRNA-control group（ P〈0. 05）. Conclusion： GRP78 functions as a positive regulator in the invasion and metastasis of bladder cancer cells,and may be involved in the expression of MMP2,MMP9 and Tipm-2.
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2015年第11期1490-1493,共4页
Chinese Journal of Immunology
基金
高等学校博士学科点专项科研基金(No.20125503110012)
