摘要
目的:探讨S100A6调控磷脂酰肌醇3-激酶/蛋白激酶B(PI3K/Akt)信号通路对黑色素瘤细胞(A375)增殖、侵袭和迁移影响。方法:首先将A375细胞分为4组:空白对照组(正常培养)、空载体对照组(转染pcDNA3.0空载体质粒)、S100A6转染组(转染pcDNA3.0-S100A6质粒)、S100A6转染+抑制剂组(转染pcDNA3.0-S100A6质粒与PI3K抑制剂LY294002)。采用实时荧光定量PCR(RT-qPCR)法检测各组细胞S100A6 mRNA相对表达量;MTT法检测各组细胞增殖活力;Transwell检测各组细胞侵袭能力;划痕线法检测各组细胞的迁移能力;western blotting检测A375各组细胞PI3K/Akt信号通路相关分子PI3K、pPI3K、GSK3β、Akt、p-Akt蛋白及c-Myc、CyclinD1、MMP-2、MMP-9蛋白表达水平。结果:与空白对照组、空载体对照组相比,S100A6转染组细胞中S100A6 mRNA表达水平显著升高(P<0.05),细胞存活率、侵袭细胞数和迁移细胞数均显著增高(P<0.05),细胞中p-PI3K/PI3K、p-Akt/Akt、GSK3β、c-Myc、CyclinD1、MMP-2、MMP-9蛋白表达水平均显著升高(P<0.05);与S100A6转染组相比,S100A6转染+抑制剂组细胞中S100A6 m RNA表达水平无显著变化(P>0.05),细胞存活率、侵袭细胞数和迁移细胞数均显著降低(P<0.05),细胞中p-PI3K/PI3K、p-Akt/Akt、GSK3β、c-Myc、CyclinD1、MMP-2、MMP-9蛋白表达水平均显著降低(P<0.05)。结论:S100A6可促进黑色素瘤细胞A375增殖、侵袭和迁移,其机制可能是通过激活PI3K/Akt信号通路实现的。
Objective:To investigate the effects of S100 A6 on the proliferation,invasion and migration of melanoma cells(A375) by regulating phosphatidylinositol 3-kinase/protein kinase B(PI3 K/Akt) signaling pathway.Methods:A375 cells were divided into four groups:blank control group,empty vector control group,S100 A6 transfection group(transfected with pcDNA3.0-S100 A6 plasmid),S100 A6 transfection + inhibitor group(transfected with pcDNA3.0-S100 A6 plasmid and treated with PI3 K inhibitor LY294002).RT-qPCR was used to detect the relative expression of S100 A6 mRNA.The proliferation,invasion and migration of cells were detected by MTT assay,cell scratch test and Transwell assay,respectively.The protein expression levels of PI3 K,p-PI3 K,GSK3β,Akt,p-Akt,c-Myc,CyclinD1,MMP-2,MMP-9 were detected by Western blotting.Results:Compared with blank control group and empty vector control group,the S100 A6 mRNA level,cell survival rate,and the number of invasive and migrated cells were significantly increased,while the protein expression levels of p-PI3 K/PI3 K,p-Akt/Akt,GSK3β,c-Myc,CyclinD1,MMP-2 and MMP-9 were significantly increased after transfection with S100 A6(P<0.05).There was no significant change in S100 A6 mRNA expression(P>0.05),while the cell survival rate,the number of invasive and migrated cells,and the protein expression levels of p-PI3 K/PI3 K,p-Akt/Akt,GSK3β,c-Myc,CyclinD1,MMP-2 and MMP-9 were decreased in S100 A6 transfection + inhibitor group compared with S100 A6 transfection group (P<0.05).Conclusion:S100 A6 can promote the proliferation,invasion and migration of melanoma A375 cells,which may be achieved by activating PI3 K/Akt signaling pathway.
作者
刘硕
常谨
刘英琦
Liu Shuo;Chang Jin;Liu Yingqi(Department of Maxillofacial Surgery,The First Hospital of Handan,Handan 056002,China;Department of Plastic Surgery,The Affiliated Hospital of Hebei University of Engineering,Handan 056000,China)
出处
《广西医科大学学报》
CAS
2021年第11期2090-2096,共7页
Journal of Guangxi Medical University
基金
河北省2020年度医学科学研究课题计划项目(No.20200594)。
