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目的:研究和厚朴酚对膀胱癌细胞的生长抑制作用。方法:采用人类膀胱癌细胞BIU87,以和厚朴酚进行细胞毒性实验,设定不同药物浓度,经过24、48、72h药物作用,观察细胞生长和死亡状态;药物作用24h,进行流式细胞术检测,观察细胞在不同药物浓度作用下的细胞生长周期变化;细胞经过药物作用72h,观察细胞在不同药物浓度作用后克隆细胞团数量;细胞加入荧光标记物,选取不同浓度药物各作用1h,采用流式细胞仪检测该药物是否促进细胞产生氧自由基以杀伤肿瘤细胞;提取经不同药物浓度作用后的细胞蛋白质,采用蛋白质印迹法检测细胞凋亡蛋白和信号通路的蛋白表达变化情况。结果:该药物对细胞有明显的生长抑制作用;随着浓度的增高,细胞G1期和SubG1期比例逐渐增高,细胞克隆团数量呈递减趋势;细胞在药物作用下有明显的Caspase3、PARP、CyclinD1、Cdk2、P21、P27、Bax/Bcl-2、Bcl-xL的趋势性表达,同时和厚朴酚促进细胞产生活性氧自由基,激活细胞内MAPK信号通路的转导,导致肿瘤细胞死亡。结论:和厚朴酚对人类膀胱癌细胞BIU87具有明显的生长抑制作用,该药可使细胞生长在G1期停滞并诱导细胞凋亡,并通过产生活性氧自由基,激活MAPK信号通路相关蛋白的表达,抑制肿瘤细胞生长。
Objective: To study the inhibitory effect of honokiol on the growth of bladder cancer cells. Methods: Human bladder cancer cell line BIU87 was used in this study. The cytotoxicity of honokiol was used to determine the concentration of different drugs. After 24, 48 and 72 hours, the cell growth and death status were observed. The cells were treated with flow cytometry The changes of cell cycle were observed under different concentrations of different drugs. The cells were observed for the number of cells cloned after different concentrations of drugs for 72 hours. Fluorescent labels were added to the cells and different concentrations of drugs were added for 1 hour. Flow cytometry was used to detect whether the drug could promote the production of oxygen free radicals to kill the tumor cells. The cell proteins were extracted by different drug concentrations. The protein expression of apoptosis proteins and signal pathways were detected by Western blotting. Results: The drug significantly inhibited the growth of cells. With the increase of concentration, the proportion of cells in G1 phase and SubG1 phase increased gradually, and the number of cell clones decreased gradually. The cells under the action of drugs had obvious Caspase3, PARP, CyclinD1 , And the expression of Cdk2, P21, P27, Bax / Bcl-2 and Bcl-xL, and honokiol promoted the production of reactive oxygen species and activated the transduction of intracellular MAPK signaling pathway, leading to tumor cell death. CONCLUSION: Honokiol can significantly inhibit the growth of human bladder cancer cell line BIU87. It can arrest cell growth in G1 phase and induce apoptosis. By activating reactive oxygen species, honokiol activates MAPK pathway-related protein Expression, inhibition of tumor cell growth.