紫铆花素对人黑色素瘤细胞A375与人永生化角质形成细胞HaCaT共培养细胞生物学活性的影响  

Effect of Butein on the Biological Activity of Co-Cultured Human Melanoma Cells A375 and Human Immortalized Keratinocytes HaCaT

作　　者：高莉 谭雪 罗静莺 闫明 GAO Li;TAN Xue;LUO Jingying;YAN Ming(Key Laboratory of Traditional Uyghur Medicine Prescription,Institute of Xinjiang Traditional Uyghur Medicine,Urumqi,Xinjiang,China 830011)

机构地区：[1]新疆维吾尔自治区维吾尔医药研究所新疆维吾尔医方剂学重点实验室,新疆乌鲁木齐830011

出　　处：《中国药业》2024年第6期36-39,共4页China Pharmaceuticals

基　　金：国家自然科学基金[81260689];新疆维吾尔自治区自然科学基金重点项目[2022D01D25]。

摘　　要：目的探讨紫铆花素对人黑色素瘤细胞A375与人永生化角质形成细胞HaCaT共培养细胞生物学活性的影响。方法建立A375细胞与HaCaT细胞的共培养细胞体系,实验分为对照组(等体积培养液),紫铆花素低、中、高剂量组(以下简称低、中、高剂量组,0.5,1.0,5.0μg/L)。采用四甲基偶氮噻唑蓝(MTT)法检测细胞活性;采用流式细胞仪检测细胞的周期分布和线粒体膜电位;采用实时荧光定量聚合酶链反应(qPCR)法检测干细胞因子(SCF)和其受体c-Kit,以及小眼畸形相关转录因子(MITF)的mRNA表达水平;采用Western blot法检测细胞酪氨酸酶相关蛋白1,2(TRP-1,TRP-2)及SCF,c-Kit,MITF的蛋白表达水平。结果与对照组比较,中、高剂量组细胞的增殖率均显著升高(P<0.01);与对照组比较,各剂量组G0/G1期细胞比例均显著降低,S期和G_(2)/M期细胞比例均显著升高(P<0.01),高线粒体膜电位细胞比例均显著升高(P<0.05或P<0.01),SCF,c-Kit,MITF的mRNA表达水平及SCF,c-Kit,MITF,TRP-1,TRP-2的蛋白表达水平均显著升高(P<0.05或P<0.01)。结论紫铆花素可能通过调节SCF,cKit,MITF的表达而影响A375细胞与HaCaT细胞共培养细胞的增殖、细胞周期和线粒体膜电位等细胞生物学活性。Objective To investigate the effect of butein on the biological activity of co-cultured human melanoma cells A375 and human immortalized keratinocytes HaCaT.Methods Co-cultured A375 and HaCaT cells were established and divided into the control group(equal volume of culture medium),the butein low-,medium-and high-dose groups(hereinafter referred to as the low-,medium-and high-dose groups,0.5,1.0,5.0μg/L).The cell activity was detected by the methyl thiazolyl tetrazolium(MTT)method.The cell cycle and mitochondrial membrane potential were detected by the flow cytometry.The expression levels of stem cell factor(SCF),its receptor c-Kit,microphthalmia-associated transcription factor(MITF)mRNA were detected by the real-time fluorescence quantitative polymerase chain reaction(qPCR).The expression levels of tyrosinase-related protein-1(TRP-1),tyrosinase-related protein-2(TRP-2),SCF,c-Kit and MITF proteins in cells were detected by the Western blot.Results Compared with that in the control group,the proliferation rate in the medium-and high-dose groups significantly increased(P<0.01).Compared with those in the control group,the proportion of cells at G0/G1 phase in the low-,medium-and high-dose groups significantly decreased,while the proportion of cells at S and G_(2)/M phases significantly increased(P<0.01);the proportion of cells with high mitochondrial membrane potential significantly increased(P<0.05 or P<0.01);the expression levels of SCF,c-Kit,MITF mRNA and the expression levels of SCF,c-Kit,MITF,TRP-1 and TRP-2 proteins significantly increased(P<0.05 or P<0.01).Conclusion Butein may affect the proliferation,cell cycle and mitochondrial membrane potential of co-cultured A375 and HaCaT cells by regulating the expression of SCF,c-Kit and MITF.

关 键 词：紫铆花素 人黑色素瘤细胞A375 人永生化角质形成细胞HaCaT 细胞周期 线粒体膜电位 干细胞因子 干细胞因子受体 小眼畸形相关转录因子 

分 类 号：R965[医药卫生—药理学] R285[医药卫生—药学]