槐定碱通过靶向下调β-Catenin抑制前列腺癌PC3细胞的增殖与侵袭  

Effect of sophoridine on the growth and invasion of prostate cancer PC-3 cells via down-regulation ofβ-Catenin

作　　者：马俊 陆祺中 韩玮 余琪伟 张曦[2] 姚林亚[2] 吴余凡 张犁 MA Jun;LU Qizhong;HAN Wei;YU Qiwei;ZHANG Xi;YAO Linya;WU Yufan;ZHANG Li(Department of Urology,Jiangsu Province Hospital of Traditional Chinese Medicine,Affiliated Hospital of Nanjing University of Chinese Medicine,Jiangsu Province,Nanjing210029,China;Department of Urology,Kunshan Hospital of Traditional Chinese Medicine,Jiangsu Province,Kunshan215300,China;Department of General Surgery,Kunshan First People’s Hospital,Jiangsu Province,Kunshan215300,China)

机构地区：[1]南京中医药大学附属医院江苏省中医院泌尿外科,江苏南京210029 [2]江苏省昆山市中医医院泌尿外科,江苏昆山215300 [3]江苏省昆山市第一人民医院普外科,江苏昆山215300

出　　处：《中国医药导报》2024年第20期30-35,共6页China Medical Herald

基　　金：江苏省昆山市科技计划项目(KS18035)。

摘　　要：目的探讨槐定碱通过靶向下调β-Catenin从而对前列腺癌PC3细胞增殖与侵袭的影响。方法选取人前列腺癌PC3细胞,经CCK8实验(10、20、40、80、160、320、640 mg/L槐定碱处理)检测增殖能力改变情况,计算出IC_(50),并取3个适合的作用浓度(50、100、200 mg/L组)处理细胞,以未加药处理为对照组,分别作用PC3细胞48 h后,细胞划痕实验检测迁移能力,Transwell侵袭实验检测侵袭能力;应用蛋白质印迹法检测β-Catenin、Cyclin D1和基质金属蛋白酶(MMP-9)等Wnt/β-Catenin通路内关键蛋白的水平。通过分子对接预测槐定碱与β-Catenin相互作用。结果CCK8实验表明,与对照组比较,10、20、40、80、160、320、640 mg/L槐定碱处理的PC3细胞抑制率升高(P<0.01);与对照组比较,50、100、200 mg/L组PC3细胞迁移与侵袭能力下降(P<0.05),100、200 mg/L组PC3细胞β-Catenin、CyclinD1及MMP-9蛋白下调(P<0.05);与50 mg/L组比较,100、200 mg/L组PC3细胞迁移能力、侵袭能力及β-Catenin和CyclinD1蛋白水平下降(P<0.05),200 mg/L组MMP-9蛋白下调(P<0.05);与100组比较,200 mg/L组PC3细胞迁移能力、侵袭能力及β-Catenin和MMP-9蛋白水平下降(P<0.05)。经分子对接预测分析,槐定碱可能与β-Catenin蛋白上的Gln407、Lys233、Arg469、Lys180、Ser411和Asp412等多个位点发生相互作用。结论槐定碱可能与β-Catenin蛋白相互作用,抑制PC3细胞内β-Catenin、CyclinD1及MMP-9蛋白的表达,从而影响Wnt/β-Catenin通路活性,抑制癌细胞的生长与侵袭,故槐定碱在前列腺癌中起到抑癌作用。Objective To investigate the effect of sophoridine on the proliferation and invasion of prostate cancer PC3 cells via down-regulation ofβ-Catenin.Methods PC3 cells of human prostate cancer were selected,the change of proliferation capacity was detected by CCK8 assay(10,20,40,80,160,320,640 mg/L sophoridine treatment),and IC_(50) was calculated,and 3 cells treated with appropriate concentration were set into 50,100 and 200 mg/L groups,and no drug treatment was used as the control group.Cell scratch assay was used to detect the migration ability of each group,Transwell assay was used to detect the invasion ability of each group.The levels of key proteins in Wnt/β-Catenin pathway,such asβ-Catenin,Cyclin D1 and MMP-9,were detected by Western blotting.The interaction between sophoridine andβ-Catenin was predicted by molecular docking.Results CCK8 experiment showed that the inhibition rate of PC3 cells treated with 10,20,40,80,160,320,640 mg/L sophoridine was increased compared with the control group(P<0.01).Compared with the control group,the migration and invasion ability of PC3 cells in 50,100 and 200 mg/L groups were decreased(P<0.05),and the proteins ofβ-Catenin,CyclinD1 and MMP9 in 100 and 200 mg/L groups were down-regulated(P<0.05).Compared with 50 mg/L group,migration ability,invasion ability,β-Catenin and CyclinD1 protein levels of PC3 cells in 100 and 200 mg/L groups were decreased(P<0.05),and MMP9 protein levels in 200 mg/L group were increased(P<0.05).Compared with 100 group,migration ability,invasion ability,β-Catenin and MMP9 protein levels of PC3 cells in 200 mg/L group were decreased(P<0.05).By molecular docking prediction analysis,sophoridine may interact with Gln407,Lys233,Arg469,Lys180,Ser411 and Asp412 onβ-Catenin protein.Conclusion Sophoridine may interact withβ-Catenin protein to inhibit the expression ofβ-Catenin,CyclinD1 and MMP9 proteins in PC3 cells,thereby affecting the activity of Wnt/β-Catenin pathway and inhibiting the growth and invasion of cancer cells.Therefore,sophoridine pl

关 键 词：槐定碱 前列腺癌 增殖 迁移 侵袭 

分 类 号：R737[医药卫生—肿瘤]