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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 the effect of silencing GRP78 / BIP expression in vitro on invasion and migration of bladder cancer T24 cells and its possible mechanism. METHODS: Small interfering RNA targeting GRP78 gene was designed and synthesized and transfected into bladder cancer T24 cells by Lipofectamine mine RNAIMAX. Real-time PCR and Western blot were used to detect GRP78, MMP2, MMP9 and Timp -2 expression. The effects of silencing GRP78 expression on the invasion and migration of bladder cancer T24 cells were detected by Transwell assay and cell scratch assay. Results: The expression of GRP78 was significantly decreased in T24 cells (siRNA-GRP78), the invasion and migration of cells were significantly inhibited, the expressions of MMP2 and MMP9 were decreased, while the expressions of Timp-2 were significantly increased (all P <0.05). Conclusion: Silencing GRP78 can significantly inhibit the invasion and migration of human bladder cancer cells. The mechanism may be related to the effects of MMP-2, MMP9 and Timp-2 expression.