CXCR4抑制剂对膀胱癌细胞的增殖、迁移和侵袭的影响及其机制的研究

CXCR4 regulates the development of bladder cancer via wnt/β-catenin signaling pathway

目的探讨趋化因子受体4(CXCR4)在膀胱癌发展过程中的作用及其调控机制。方法采用qPCR法检测膀胱癌细胞株SW780、5637和T24中CXCR4mRNA的表达。在SW780细胞加入CXCR4抑制剂AMD346510uM处理后,以未加入AMD3465的SW780细胞为对照组,用MTT、Transwell法分别检测抑制CXCR4对膀胱癌细胞增殖、迁移和侵袭性的作用。Westernblotting检测β-catenin、Wnt/β-catenin通路相关基因和EMT相关基因的表达变化。用裸鼠皮下异体移植模型研究CXCR4对人膀胱癌SW780细胞裸鼠移植瘤生长的影响。结果SW780膀胱癌细胞经AMD3465处理48、72h后,细胞增殖减慢,其吸光度值与对照组相比差异有统计学意义(P<0.001)。经AMD3465处理48h后的SW780膀胱癌细胞迁移指数、侵袭指数降低,与对照组相比差异有统计学意义(P<0.001)。AMD3465明显抑制β-catenin、Wnt/β-catenin通路相关基因MMP-2和c-myc的蛋白表达,同时上调E-cadherin的蛋白表达。结论抑制CXCR4可以通过调控Wnt/β-catenin通路抑制膀胱癌的进展。

Objective To investigate the role of chemokine receptor 4 (CXCR4) in the development of bladder cancer. Methods QPCR was used to detect the expression of CXCR4 mRNA in normal bladder tissues, bladder cancer tissues and bladder cancer cell lines SW780, 5637 and T24. MTT, colony formation, and transwell migration and invasion assays were performed in SW780 cells after treatment with the CXCR4 antagonist AMD3465.β-catenin, Wnt/β-catenin pathway targeted genes and epithelial-mesenchymal transition (EMT) related genes were detected by Western blot. The effect of AMD3465 on the growth of human bladder cancer SW780 cell xenografts was evaluated using a nude mice model. Results The expression of CXCR4 in bladder cancer and bladder cancer cell lines was significantly higher than that in normal bladder tissues. AMD3465 could effectively inhibit the proliferation, colony formation, migration, and invasion of SW780 cells, meanwhile, it significantly inhibited the expression of β-catenin and Wnt/β-catenin pathway targeted genes. The expression of EMT related gene E-cadherin was significantly up-regulated. Conclusions CXCR4 affects the development of bladder cancer by regulating Wnt/β-catenin signaling pathway.