桧木醇激活膀胱癌J82细胞自噬诱导细胞凋亡

Hinokitiol induces bladder cancer J82 cells apoptosis via autophagy induction

背景与目的:膀胱癌是我国最常见的泌尿系统恶性肿瘤,近年来发病率逐年上升,严重威胁着人类的健康。本研究旨在探讨桧木醇对人膀胱癌J82细胞增殖、凋亡及自噬的作用,并初步阐明其作用机制。方法:采用CCK-8法检测细胞的增殖活力,采用流式细胞术检测细胞的凋亡状态,采用蛋白[质]印迹法(Western blot)检测cleaved caspase 3、LC3及P62蛋白的表达,转染EGFP-LC3后在激光共聚焦显微镜下观察细胞自噬状态。结果:桧木醇能够显著抑制J82细胞的增殖活力,通过激活caspase途径诱导肿瘤细胞凋亡,Z-VAD-FMK能够部分抑制桧木醇的凋亡诱导作用。桧木醇能够激活J82细胞发生自噬,上调LC3蛋白的表达,下调P62蛋白的表达。3-MA抑制自噬之后能够部分逆转桧木醇的抗肿瘤作用。结论:桧木醇可以显著抑制J82细胞增殖并通过过度激活自噬促进膀胱癌细胞凋亡。

Background and purpose： Bladder cancer is the most common urological malignancy in our country which seriously threatens human health, and its incidence increased year by year. This study aimed to investigate the effects of hinokitiol on the proliferation, apoptosis and autophagy in human bladder cancer cell lines. Methods：CCK-8 assays were performed to analyze the effects of hinokitiol on the proliferation of J82 cells. Apoptosis rate was determined by lfow cytometry. Cleaved caspase 3, LC3 and P62 protein expression was determined using Western blot. EGFP-LC3 microscopy assay was performed to assess autophagy. Results： Hinokitiol significantly inhibited the proliferation of J82 cells and induced cell apoptosis via caspase pathway. The apoptosis effect of hinokitiol could be partially antagonized by Z-VAD-FMK. Hinokitiol induced autophagy of J82 cells, increased LC3 expression and down-regulated P62 expression. 3-MA is able to rescue Tet-induced cell death. Conclusion：Hinokitiol can inhibit the proliferation of J82 cells and induce cell apoptosis via autophagy activation.