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目的探讨肿瘤抑制因子CYLD提高膀胱癌细胞吉西他滨化疗敏感性。方法构建CYLD过表达质粒转染的人膀胱癌细胞株T24、253J,将细胞分为仅加入脂质体的对照组,转染空载质粒的空载组和CYLD组。Transw ell实验检测CYLD对膀胱癌细胞迁移/侵袭能力的影响,CCK8法检测膀胱癌细胞对吉西他滨的化疗敏感性,Western blotting、吖啶橙染色、电镜等检测自噬蛋白表达水平。结果与对照组和空载组相比,CYLD组通过Transw ell小室的细胞数目明显减少(P<0.05),24 h半抑制浓度(IC50)明显降低(P<0.05),自噬相关蛋白LC3、Beclin 1表达降低,p62表达增高,吖啶橙染色及电镜观察发现自噬溶酶体及自噬泡减少。结论肿瘤抑制因子CYLD可以抑制膀胱癌细胞的迁移/侵袭能力,并通过抑制自噬提高膀胱癌细胞对吉西他滨的化疗敏感性。
Objective To investigate the chemosensitivity of tumor suppressor CYLD to gemcitabine in bladder cancer cells. Methods The human bladder cancer cell line T24,253J transfected with CYLD overexpression plasmid was constructed. The cells were divided into control group, which was only added liposome, and transfected into empty vector and CYLD group. Transw ell assay was used to detect the effect of CYLD on the migration and invasion of bladder cancer cells. CCK8 assay was used to detect the chemosensitivity of bladder cancer cells to gemcitabine. Western blotting, acridine orange staining and electron microscopy were used to detect the expression of autophagy. Results Compared with the control group and the no-load group, the number of cells passing through the Transwell cell in CYLD group was significantly decreased (P <0.05), the IC50 at 24 h was significantly decreased (P <0.05), the expression of autophagy-related proteins LC3, Beclin 1 expression decreased, p62 expression increased, acridine orange staining and electron microscopy found that autophagy lysosomes and autophagy decreased. Conclusion The tumor suppressor CYLD can inhibit the migration / invasion ability of bladder cancer cells and enhance the chemosensitivity of bladder cancer cells to gemcitabine by inhibiting autophagy.