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目的探讨成纤维细胞生长因子受体2(FGFR2)的剪切突变体FGFR2Ⅲc对多柔比星耐药的膀胱癌细胞间质-上皮转化的调节作用。方法通过浓度递增法建立人膀胱癌253J细胞系的多柔比星耐药253J/DOX细胞株。分别用MTT实验、Western blot法和实时定量PCR(qRT-PCR)比较253J细胞和253J/DOX细胞对多柔比星的敏感性、P-糖蛋白表达水平和FGFR2Ⅲc表达水平的差异。Western blot法检测253J细胞和253J/DOX细胞E-cadherin和vimentin表达。划痕愈合实验比较253J细胞和253J/DOX细胞迁移能力的差异。结果与亲代细胞相比,253J/DOX细胞P-糖蛋白表达上调并对多柔比星明显耐药(P<0.05)。FGFR2Ⅲc mRNA在253J/DOX细胞的表达明显高于253J细胞。同时,与亲代细胞相比,253J/DOX细胞E-cadherin表达上调,vimentin表达下调,体外迁移能力下降。结论 FGFR2Ⅲc在化疗耐药的膀胱癌细胞表达上调,可能通过诱导间质-上皮转化促进肿瘤迁移灶形成。
Objective To investigate the regulatory effect of FGFR2Ⅲc, a FGFR2-cut mutant, on the stromal-epithelial transformation of multidrug-resistant bladder cancer cells. Methods Doxorubicin resistant 253J / DOX cell line was established by increasing concentration method. The sensitivity, the expression of P-glycoprotein and the expression level of FGFR2Ⅲc in 253J cells and 253J / DOX cells were compared by MTT assay, Western blot and real-time quantitative PCR (qRT-PCR). Western blot was used to detect the expression of E-cadherin and vimentin in 253J and 253J / DOX cells. Scratch healing experiments compared 253J cells and 253J / DOX cell migration ability differences. Results Compared with parental cells, the expression of P-glycoprotein in 253J / DOX cells was up-regulated and resistant to doxorubicin (P <0.05). The expression of FGFR2Ⅲc mRNA in 253J / DOX cells was significantly higher than that of 253J cells. Meanwhile, compared with parental cells, E-cadherin expression in 253J / DOX cells was up-regulated, while vimentin expression was down-regulated and migration in vitro was decreased. Conclusion FGFR2Ⅲc is up-regulated in chemoresistant bladder cancer cells, which may promote the formation of tumor metastases by inducing the interstitial-epithelial transformation.