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目的:研究土槿皮乙酸对人肾癌细胞A498凋亡的影响,并探讨其内在的分子机制,为肾癌治疗寻找有效的新靶点和新策略。方法:人肾癌细胞A498经10、15μmol/L土槿皮乙酸处理48 h后,用流式细胞仪检测细胞凋亡,同时通过Western印迹和实时荧光定量PCR检测凋亡相关蛋白和m RNA的表达;加入PI3K/Akt通路抑制剂LY294002或15μmol/L土槿皮乙酸处理A498细胞48 h后,Western印迹检测相关蛋白的表达。结果:经不同浓度土槿皮乙酸作用A498细胞48 h后,与未加土槿皮乙酸组细胞相比,细胞凋亡率显著上升,且呈剂量依赖性,比较差异有统计学意义(P<0.05);同时,Blc-2表达减少,Bax、caspase-9、caspase-3表达增多。Blc-2蛋白的表达量随LY294002或土槿皮乙酸的逐步加入而减少,而Bax、caspase-9、caspase-3的表达呈增加趋势;PI3K和Akt蛋白的表达量与是否加入LY294002或土槿皮乙酸无关,p-PI3K和Aktp-Ser473蛋白的表达量随LY294002或土槿皮乙酸的逐步加入而减少。结论:土槿皮乙酸可抑制A498细胞凋亡,其作用机制可能与PI3K/Akt通路相关。
OBJECTIVE: To study the effect of hyaluronic acid on the apoptosis of human renal cell carcinoma A498 and to explore its intrinsic molecular mechanism, so as to find a new target and a new strategy for the treatment of renal cell carcinoma. METHODS: Human renal carcinoma cell line A498 was treated with 10, 15μmol / L hyaluronic acid for 48 hours, then the apoptosis was detected by flow cytometry. Apoptosis-related proteins and m RNA were detected by Western blotting and real-time fluorescence quantitative PCR A498 cells were treated with PI3K / Akt pathway inhibitor LY294002 or 15μmol / L hyaluronic acid for 48 hours, Western blotting was used to detect the expression of related proteins. Results: The apoptosis rate of A498 cells treated with acetic acid at different concentrations for 48 h was significantly increased compared with that of the cells without acetic acid (P <0.05), and the apoptosis rate was significantly increased in a dose - dependent manner (P < 0.05). Meanwhile, the expression of Bcl-2, Bax, caspase-9 and caspase-3 were increased. The expression of Blc-2 protein decreased with the gradual addition of LY294002 or hyaluronic acid, while the expression of Bax, caspase-9 and caspase-3 showed an increasing trend. The expression of PI3K and Akt protein increased with the increase of LY294002 or Hibiscus P <0.01). The expression of p-PI3K and Aktp-Ser473 protein decreased with the gradual addition of LY294002 or hyaluronic acid. CONCLUSION: Hyaluronic acid inhibits the apoptosis of A498 cells and its mechanism may be related to the PI3K / Akt pathway.