摘要
目的:探讨中性乳酸对肝癌细胞增殖、迁移及侵袭能力的影响,及其可能的作用机制。方法:肝癌SMMC-7721和HepG2细胞分别予以不同浓度(0、2.5、5和10 mmol/L)(以0 mmol/L浓度组为对照组)的中性乳酸处理后,采用MTT法检测细胞的增殖能力;Transwell小室法检测细胞的迁移和侵袭能力;蛋白质印迹法检测各组肝癌细胞中磷脂酰肌醇3-激酶(phosphoinositide 3-kinase,PI3K)、蛋白激酶B(protein kinase B,PKB,又称Akt)、磷酸化Akt(Ser473)[phospho-Akt(Ser473),p-Akt(S473)]、基质金属蛋白酶2(matrix metallopeptidase-2,MMP-2)和MMP-9蛋白的表达水平。将SMMC-7721和HepG2细胞各分为4组,分别予以等体积培养液(对照组)、中性乳酸、PI3K/Akt信号通路抑制剂哌立福新和中性乳酸联合哌立福新处理后,再用MTT法和Transwell小室实验检测各组肝癌细胞的增殖、迁移和侵袭能力,蛋白质印迹法检测细胞中PI3K、Akt、p-Akt(S473)、MMP-2和MMP-9蛋白的表达水平。结果:与对照组相比,5和10 mmol/L中性乳酸处理组SMMC-7721和HepG2细胞的增殖、迁移和侵袭能力均明显增强(P值均<0.01);2株细胞中PI3K、p-Ak(tS473)、MMP-2和MMP-9蛋白的表达水平均明显上调(P值均<0.01)。与对照组相比,哌立福新单药组SMMC-7721和HepG2细胞的增殖、迁移和侵袭能力均明显降低(P值均<0.05),且PI3K、p-Akt(S473)、MMP-2和MMP-9蛋白表达水平均明显下调(P值均<0.01);与中性乳酸单药组相比,中性乳酸联合哌立福新处理组SMMC-7721和HepG2细胞增殖、迁移和侵袭能力均明显降低(P值均<0.01),且PI3K、p-Akt(S473)、MMP-2和MMP-9蛋白的表达水平均明显下调(P值均<0.01)。结论:中性乳酸通过激活PI3K/Akt信号通路促进肝癌细胞的增殖、迁移和侵袭,PI3K/Akt抑制剂哌立福新可以显著减少中性乳酸对肝癌细胞增殖、迁移和侵袭的促进作用。
Objective:To investigate the effects of neutral lactate on the proliferation,migration and invasion of hepatocellular carcinoma(HCC)cells,and to explore its possible molecular mechanism.Methods:The HCC SMMC-7721 and HepG2 cells were treated with 0(as the control),2.5,5 and 10 mmol/L neutral lactate,respectively.Then the proliferation of HCC cells was detected by MTT assay.The migration and invasion abilities of HCC cells were detected by Transwell chamber method.The expression levels of phosphoinositide 3-kinase(PI3K),protein kinase B(PBK,Akt),phospho-Akt(Ser473)[p-Akt(S473)],matrix metallopeptidase-2(MMP-2)and MMP-9 were detected by Western blotting.The HCC cells SMMC-7721 and HepG2 were divided into four groups,and treated with equal volume of medium(as the control),neutral lactate,perifosine(an inhibitor of PI3K/Akt pathway),and neutral lactate combined with perifosine,respectively.Then MTT assay and Transwell chamber assay were used to detect the proliferation,migration and invasion abilities of HCC cells in each group.The expression levels of PI3K,Akt,p-Akt(S473),MMP-2 and MMP-9 proteins in HCC cells were detected by Western blotting.Results:The proliferation,migration and invasion abilities of SMMC-7721 and HepG2 cells in 5 and 10 mmol/L neutral lactate treatment groups were significantly enhanced as compared with the control group(all P<0.01).The expression levels of PI3K,p-Akt(S473),MMP-2 and MMP-9 proteins in HCC cells treated with neutral lactate were significantly increased as compared with the control group(all P<0.01).Compared with the control group,the proliferation,migration and invasion abilites of HCC cells in the perifosine treatment group were significantly decreased(all P<0.05),and the expression levels of PI3K,p-Akt(S473),MMP-2 and MMP-9 proteins were also significantly down-regulated(all P<0.05).Compared with the neutral lactate treatment group,the proliferation,migration and invasion abilities of HCC cells in the neutral lactate combined with perifosine treatment group were significantly re
作者
彭明兵
郑志菊
曾智锐
张强
孙达权
夏庆
徐国强
PENG Mingbing;ZHENG Zhiju;ZENG Zhirui;ZHANG Qiang;SUN Daquan;XIA Qing;XU Guoqiang(School of Basic Medical Sciences,Guizhou Medical University,Guiyang 550025,Guizhou Province,China;Operating Room,Second People’s Hospital of Guiyang,Guiyang 550023,Guizhou Province,China;Scientific Research Center,Third Affiliated Hospital of Guizhou Medical University,Duyun 558000,Guizhou Province,China)
出处
《肿瘤》
CAS
CSCD
北大核心
2020年第1期9-19,共11页
Tumor
基金
贵州省中医药、民族医药科学技术研究专项课题资助项目(编号:QZYY-2015-012)
贵州省卫生计生委科学技术基金资助项目(编号:gzwjkj2014-1-029).
