FGFR2IIIc在膀胱癌化疗耐药细胞株中的表达及意义

Expression and significance of FGFR2Ⅲc in chemoresistant bladder cancer cells

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 role of flbroblast growth factor receptor-2 （FGFFI2） splice variant FGFR219c in the regulation of mesenchymal-epithelial transition （MET） in doxorubicin-resistant bladder cancer cells. Methods A doxorubicin- resistant human bladder cell line （253J/DOX） was generated from the bladder cancer cell line 253J by being continuously exposed to gradually increasing doses of doxorubicin. ChemosensitMty to doxorubicin was determined by MTT assay. The expressions of P-glycoprotein and FGFR2 Ⅲ c were evaluated by Western blotting and real-time RT-PCR, respectively. Changes in E-cadhedn and vimentin were detected by Western blot analysis. Migration ability of 253J and 253J/DOX cells was analyzed by in vitro wound healing assay. Results The resistant cells, 253J/DOX, were more resistant to doxorubicin than the parent cells. Western blotting and RT-PCR analysis indicated the higher levels of P-glycoprotein and FGFR2 19 c in 253J/DOX cells （P 〈 0.05）. Additionally, compared with the 253J cells, 253J/DOX cells presented the upregulation of E-cadherin, the downregulation of vimentin and the inhibition of migration ability. Conclusion FGFR219 c-induced MET in chemoresistant bladder cancer cells may play an important role in the formation of metastatic lesions.