川芎嗪对人皮肤恶性黑素瘤A875细胞增殖和凋亡相关蛋白表达的影响  

Effects of ligustrazine on proliferation and apoptosis-related protein expression in human skin malignant melanoma cell A875

作　　者：肖敏[1] 徐洪来[2] XIAO Min;XU Hong-lai(Department of Dermatology,Liuzhou People′s Hospital,Liuzhou 545006,China;Department of Hepatobiliary Surgery,Liuzhou People′s Hospital,Liuzhou 545006,China)

机构地区：[1]广西柳州市人民医院皮肤科,柳州市545006 [2]广西柳州市人民医院肝胆外科,柳州市545006

出　　处：《广西医学》2020年第14期1846-1849,共4页Guangxi Medical Journal

基　　金：广西医药卫生科研课题(Z2013215,Z2014430);广西柳州市人民医院博士硕士医学基础研究启动基金(lryjj201504,lryjj201404);广西柳州市科技计划(2018BJ10302)。

摘　　要：目的探讨川芎嗪对人皮肤恶性黑素瘤A875细胞增殖和凋亡相关蛋白表达的影响。方法体外培养A875细胞,采用不同浓度(0 mg/mL、0.5 mg/mL、1 mg/mL、2 mg/mL)川芎嗪分别作用A875细胞48 h、72 h,采用四甲基偶氮唑盐法检测各组细胞存活率。采用不同浓度川芎嗪(0 mg/mL、2 mg/mL)分别作用A875细胞0 h、48 h、72 h,采用蛋白免疫印迹法检测环氧化酶-2(COX-2)、含半胱氨酸的天冬氨酸蛋白水解酶3(Caspase3)蛋白相对表达水平,并分析经2 mg/mL川芎嗪干预48 h、72 h后两者表达水平的相关性。结果经川芎嗪干预48 h、72 h后,A875细胞的细胞存活率随着川芎嗪浓度的升高而降低(均P<0.05);当川芎嗪浓度为1或2 mg/mL时,A875细胞的细胞存活率随着药物作用时间延长而下降(均P<0.05)。2 mg/mL川芎嗪干预0 h、48 h、72 h,A875细胞中COX-2蛋白表达水平依次降低,Caspase3蛋白表达水平依次升高;与0 mg/mL川芎嗪干预比较,应用2 mg/mL川芎嗪干预48 h和72 h后细胞中COX-2蛋白表达水平更低,Caspase3蛋白表达水平更高(均P<0.05)。2 mg/mL川芎嗪干预A875细胞48 h、72 h后,A875细胞的COX-2蛋白与Caspase3蛋白的相对表达水平均呈负相关(均P<0.05)。结论川芎嗪能抑制A875细胞增殖,并可抑制COX-2活化、上调Caspase3表达水平而促进细胞凋亡。Objective To explore the effects of ligustrazine on proliferation and apoptosis-related protein expression in human skin malignant melanoma cell A875.Methods A875 cells cultured in vitro were treated with different concentrations(0 mg/mL,0.5 mg/mL,1 mg/mL,2 mg/mL)of ligustrazine for 48 and 72 hours,respectively,and methyl thiazolyl tetrazolium assay was employed to detect cell survival rate in each group.A875 cells were treated with different concentrations(0 mg/mL,2 mg/mL)of ligustrazine for 0,48 and 72 hours,respectively,Western blot assay was employed to detect the relative protein expression levels of cyclooxygenase 2(COX-2)and cysteinyl aspartate specific proteinase 3(Caspase3),and the correlation between COX-2 and Caspase3 expression levels,which were intervened by 2 mg/mL for 48 or 72 hours,was analyzed.Results After 48-or 72-hour ligustrazine intervention,the cell survival rate of A875 cells decreased with the increase of ligustrazine concentration(all P<0.05);when the concentration of ligustrazine was 1 or 2 mg/mL,the cell survival rate of A875 cells decreased with the prolongation of drug action time(all P<0.05).The expression level of COX-2 protein decreased in turn while the expression level of Caspase3 protein increased in turn in A875 cells intervened by 2 mg/mL ligustrazine for 0,48 and 72 hours;by comparison with 0 mg/mL ligustrazine,the expression level of COX-2 protein was lower and the expression level of Caspase3 protein was higher after 48-and 72-hour of intervention by 2 mg/mL ligustrazine(all P<0.05).After 48-or 72-hour intervention by 2 mg/mL ligustrazine,the relative expression levels of COX-2 and Caspase3 proteins in A875 cells showed a negative correlation(all P<0.05).Conclusion Ligustrazine can inhibit A875 cell proliferation as well as inhibit COX-2 activation and up-regulate Caspase3 expression level to promote cell apoptosis.

关 键 词：恶性黑素瘤 川芎嗪 环氧化酶-2 含半胱氨酸的天冬氨酸蛋白水解酶3 体外实验 

分 类 号：R751.05[医药卫生—皮肤病学与性病学]