乌苯美司诱导肾癌细胞死亡的作用及机制

Role and possible mechanism of autophagy in ubenimex-induced cell death of renal cell carcinoma

目的观察乌苯美司对肾癌细胞增殖及死亡的影响并探讨其可能机制。方法利用WST-8细胞增殖实验及细胞生长曲线检测乌苯美司对肾癌细胞增殖能力的影响;LDH释放实验检测乌苯美司对肾癌细胞死亡的影响;酶活性测定法及Western blotting研究乌苯美司诱导肾癌细胞APN活性及表达;Western blotting及免疫组化染色观察乌苯美司的作用靶点氨肽酶N(APN)/CD13在肾癌组织及正常组织中的表达。结果乌苯美司不能降低肾癌细胞中APN的表达但是可以抑制其活性;乌苯美司可抑制肾癌细胞增殖及活力,诱导细胞自噬性死亡。结论乌苯美司可抑制肾癌细胞增殖,并可通过诱导自噬引发肾癌细胞死亡。

Objective To determine the effect of ubenimex on the proliferation of renal cell carcinoma cells and the possible mechanism. Methods WST-8 cell proliferation assay and cell growth curve were used to detect the proliferation of renal cell carcinoma cells. LDH cytotoxicity assay was used to detect the cell death of the renal cell carcinoma. Alanine-p-nitroanilido was used to detect the activity and expression of APN induced by ubenimex in renal cell carcinoma cell lines. Western blotting and immunohistochemical staining were used to detect the expression of APN/CD13 in tissues of normal kidney and renal cell carcinoma. Results Ubenimex could not reduce the express of APN in renal cell carcinoma but inhibit its activity. Ubenimex could inhibit the proliferation and activity of renal cell carcinoma cells, and induce autophagic cell death. Conclusion Ubenimex inhibits the proliferation of renal cell carcinoma cells and promotes levels of cell death in renal cell carcinoma cells via an autophagic mechanism.