骨形态发生蛋白-2调控MAPK信号通路在肝癌干细胞特征维持中的机制研究  

Study on the mechanism of bone morphogenetic protein-2 regulation of MAPK signaling pathway in the maintenance of hepatocellular carcinoma stem cell characteristics

作　　者：宫东伟 邢贻雷[1] 王开琼[1] 黄涛[2] 郑进方[1] GONG Dongwei;XING Yilei;WANG Kaiqiong;HUANG Tao;ZHENG Jinfang(Department of Hepatobiliary and Pancreatic Surgery,Hainan Provincial People's Hospital,Haikou,57031l,China;Department of Hepatobiliary and Pancreatic Surgery,Henan Cancer Hospital)

机构地区：[1]海南省人民医院肝胆胰外科,海口570311 [2]河南省肿瘤医院肝胆胰外科

出　　处：《中国中西医结合消化杂志》2022年第8期573-578,590,共7页Chinese Journal of Integrated Traditional and Western Medicine on Digestion

基　　金：2019年海南省基础与应用基础研究计划项目(No:2019RC373)。

摘　　要：目的:探究骨形态发生蛋白-2(BMP-2)在肝癌干细胞特征维持中的作用及其相关机制。方法:采用qRT-PCR检测肝癌患者癌组织及其癌旁组织BMP-2和CD133的表达水平。通过慢病毒将PLK-NC和PLKBMP-2转染至HepG2细胞,设空白对照组(未转染)、PLK-NC组(转染PLK-NC)、PLK-BMP-2组(转染PLKBMP-2)与PLK-BMP-2+PNU-120596组(转染PLK-BMP-2后给予5μmol/LPNU-120596孵育24h)。采用qRT-PCR检测各组细胞BMP-2、EpCAM、CD133、CD24基因表达水平,Westernblot检测BMP-2、p-MAPK蛋白表达水平。通过有限稀释法测定各组干细胞的成球频率差异。结果:与癌旁组织比较,癌组织BMP-2和CD133表达水平升高。与PLK-NC组比较,PLK-BMP-2组BMP-2、EpCAM、CD133、CD24基因表达水平升高,BMP-2、p-MAPK蛋白表达水平升高,细胞成球频率增加(均P<0.05)。与PLK-BMP-2组比较,PLK-BMP-2+PNU-120596组BMP-2表达水平差异无统计学意义(P>0.05),EpCAM、CD133、CD24基因表达水平降低(P<0.05),p-MAPK蛋白表达水平降低(P<0.05),细胞成球频率减少(P<0.05)。结论:BMP-2通过MAPK信号通路维持肝癌细胞干性,有望成为肝癌治疗的潜在靶点。Objective:To investigate the role of bone morphogenetic protein-2(BMP-2)in maintaining of hepatocllular carcinoma stem cell characteristics and its related mechanisms.Methods:The gene expression levels of BMP-2 and CD133 in liver cancer tissues and adjacent tissues were detected by qRT-PCR.HepG2 cells were transfected with PLK-NC and PLK-BMP-2 by lentivirus,and the groups were divided into blank control group,PLKNC group,PLK-BMP-2 group and PLK-BMP-2+PNU-120596 group(5μmol/L PNU-120596 after transfection).The gene expression levels of BMP-2,EpCAM,CD133 and CD24 were detected by qRT-PCR,and the protein expression levels of BMP-2 and p-MAPK were detected by Western blot.The formation spheroids frequency of stem cells in each group was determined by the limited dilution method.Results:Compared with adjacent tissue,BMP-2 and CD133 expression levels were increased in tumor tissue.Compared with PLK-NC group,the gene expression levels of BMP-2,EpCAM,CD133 and CD24,proteins expression levels of BMP-2 and p-MAPK,and formation spheroids frequency were increased in PLK-BMP-2 group(P<0.05).Compared with PLK-BMP-2 group,there was no difference in the expression level of BMP-2 in PLK-BMP-2+PNU-120596 group(P>0.05),while the genes expression level of EpCAM,CD133 and CD24,protein expression level of p-MAPK and formation sphe roids frequency were decreased(P<o.05).Conclusion:BMP-2 maintains hepatocellular carcinoma stem cell characteristics through MAPK signaling pathway and is expected to be a potential therapeutic target for hepatocellular carcinoma.

关 键 词：骨形态发生蛋白-2 肝癌 干细胞 丝裂原活化蛋白激酶 

分 类 号：R735.7[医药卫生—肿瘤]