HSP90β对人骨肉瘤MG63细胞干性特征的影响及分子影响机制  

Effects of HSP90βon the stemness characteristics of human osteosarcomaa MG63 cells and its molecular mechanism

作　　者：李增佑 舒雄[2] 刘辉琦[1] 刘永年[1] Li Zengyou;Shu Xiong;Liu Huiqi;Liu Yongnian(Medical Department of Qinghai University,Xining 810001,China;Beijing Institute of Orthopaedics and Traumatology/Jishuitan Hospital,Beijing 100035,China)

机构地区：[1]青海大学医学部,青海西宁810001 [2]北京市创伤骨科研究所/北京积水潭医院,北京100035

出　　处：《中国高原医学与生物学杂志》2023年第1期55-68,共14页Journal of Chinese High Altitude Medicine & Biology

基　　金：青海省卫生健康委员会医药卫生科技项目(2017-wjzdx-72)。

摘　　要：目的研究HSP90β对人骨肉瘤MG63细胞干性特征的影响及其相关分子机制。方法将骨肉瘤MG63细胞进行慢病毒转染并分为四组:过表达HSP90β组(HSP90β-pEZ)、过表达对照组(NEG-pEZ)、沉默HSP90β组(HSP90β-shRNA)以及沉默对照组(NEG-shRNA)。用Western Blot法测定HSP90β蛋白以及SOX2、OCT4干性蛋白的表达水平,AKT、p-AKT、mTOR、p-mTOR等AKT/mTOR信号通路相关蛋白的表达水平;用细胞迁移实验(Transwell)法测定HSP90β对骨肉瘤MG63细胞迁移和侵袭功能的影响;用甲基纤维素成球(Self-renewal assay)法测定HSP90β对骨肉瘤MG63细胞自我更新的影响。并进行相关分子机制的研究。结果与NEG-pEZ组相比,HSP90β-pEZ组的HSP90β蛋白及干性相关蛋白表达水平明显升高(P<0.05);HSP90β-pEZ组细胞自我更新、迁移及侵袭活力均增加(P<0.05)。与NEG-shRNA组相比,HSP90β-shRNA组的HSP90β蛋白及干性相关蛋白表达水平显著降低(P<0.05);NEG-shRNA组自我更新、迁移及侵袭活力均减弱(P<0.05)。与NEG-pEZ组相比,HSP90β-pEZ组p-AKT、p-mTOR等AKT/mTOR信号通路相关蛋白的表达水平明显升高(P<0.05);与NEG-shRNA组相比,HSP90β-shRNA组、p-AKT、p-mTOR等AKT/mTOR信号通路相关蛋白的表达水平明显降低(P<0.05)。结论HSP90β基因可能通过激活AKT/mTOR信号通路促进人骨肉瘤MG63细胞的自我更新、迁移及侵袭活力,并促进骨肉瘤细胞相关干性基因的表达。这也是HSP90β对人骨肉瘤MG63细胞干性特征产生影响的分子影响机制。Objective To study the effects of HSP9oβon the stem characteristics of human osteosarcoma MG63 cells and its related molecular mechanism.Methods Osteosarcoma MG63 cells were transfected with lentivir us and divided into four groups:overexpression of HSP90βgroup(HSP90β-pEZ),overexpression control group(NEG-pEZ),silence HSP90βgroup(HSP90β-shRNA)and silencing control group(NEG-shRNA).The expression levels of HSP90βprotein,SOX2 and OCT4 stem proteins,and the expression levels of AKT/mTOR signaling pathway-related proteins such as AKT,p-AKT,mTOR and p-mTOR were determined using Western Blot method.The effect of HSP90βon the migration and invasion viabilities of osteosarcoma MG63 cells was determined by Transwell experimental method.The effect of HSP90βon the self-renewal of osteosarcoma MG63 cells was determined by self-renewal assay.Results Compared with NEG-pEZ group,the expression levels of HSP90βprotein and stem-related protein in the HSP90β-pEZ group were significantly increased(P<0.05).The self-renewal,migration and invasion viabilities of HSP90β-pEZ group were also increased(P<0.05).Compared with NEG-shRNA group,the expression levels of HSP90βprotein and stem-related protein in the HSP90β-shRNA group were significantly reduced(P<0.05).The self-renewal,migration and invasion viabilities in the NEG-shRNA group were all weakened(P<0.05).Compared with NEG-pEZ group,the expression levels of AKT/mTOR signaling pathway-related proteins such as p-AKT and p-mTOR in the HSP90β-pEZ group were significantly increased(P<0.05).Compared with NEG-shRNA group,the expression levels of AKT/mTOR signaling pathway-related proteins such as p-AKT and p-mTOR in the HSP90β-shRNA group were obviously reduced(P<0.05).Conclusions HSP90βgene may promote the self-renewal,migration and invasion viabilities of human osteosarcoma MG63 cells by activating AKT/mTOR signaling pathway,and promote the expression of osteosarcoma related stem genes.This is also HSP90βmolecular mechanisms that affect the stemness of human osteosarcoma

关 键 词：骨肉瘤 HSP90β 干性特征 

分 类 号：R738.1[医药卫生—肿瘤]