基于数据挖掘及体外实验的雷公藤甲素抗肾癌分子机制的探究

Exploring the molecular mechanism of Triptolide in anti-renal cancer via data mining and in vitro experiments

目的探究雷公藤甲素抗肾癌分子机制。方法通过网络药理学与机器学习分析雷公藤甲素作用肾癌的潜在靶点,利用CCK-8法检测其对ACHN细胞活力的影响,运用划痕实验检测其对ACHN细胞迁移抑制的影响,借助流式细胞术检测其对ACHN细胞凋亡的影响,利用蛋白免疫印迹法(Western Blot)检测重要靶点蛋白的表达差异。结果网络药理学预测雷公藤甲素通过STAT3、EGFR、AKT1、BCL2、mTOR等核心靶点抑制肾癌细胞生长。CCK-8结果显示随着药物浓度升高,ACHN细胞存活率明显下降(P<0.001),10nmol/L为最佳抑制浓度。雷公藤甲素处理后,实验组迁移率下降了10.4%(P<0.01),凋亡率增加了5.08%(P<0.05)。Western Blot结果显示p-mTOR、P53、LC3I/II表达量增加,p-STAT3、P62表达量下降。结论雷公藤甲素可抑制ACHN细胞增殖和迁移,诱导肾癌细胞自噬性凋亡。

Objective To explore the molecular mechanism of triptolide against renal carcinoma.Methods The potential target of triptolide in renal carcinoma was analyzed by network pharmacology and machine learning.The effect of triptolide on ACHN cell viability was detected by CCK-8 method,the effect of triptolide on ACHN cell migration inhibition was detected by scratch test,and the effect of triptolide on ACHN cell apoptosis was detected by flow cytometry.The expression difference of important target proteins was detected by Western blot.Results Network pharmacology predicted that triptolide inhibited the growth of renal carcinoma cells by STAT3,EGFR,AKT1,BCL2,mTOR and other core targets.The results of CCK-8 showed that the survival rate of ACHN cells decreased significantly with the increase of drug concentration(P<0.001),and 10nM was the best inhibitory concentration.After triptolide treatment,the mobility of the experimental group decreased by 10.4%(P<0.01)and the apoptosis rate increased by 5.08%(P<0.05).Western Blot showed that the expression levels of p-mTOR,P53 and LC3I/II increased,while the expression levels of pSTAT3 and P62 decreased.Conclusion Triptolide can inhibit the proliferation and migration of ACHN cells and induce autophagy apoptosis of renal carcinoma cells.