HOXB7调控Wnt通路对骨肉瘤细胞上皮-间充质转化及侵袭、迁移的影响  

HOXB7 modulates Wnt signaling to regulate epithelial-mesenehymal transition and metastasis in osteosarcoma cells

作　　者：丁木亮[1] 王俊杰[1] DING Muliang;WANG Junjie(Department of Orthopedics,the Second Xiangya Hospital of Central South University,Changsha 410011,Hunan,China)

机构地区：[1]中南大学湘雅二医院骨科,湖南长沙410011

出　　处：《贵州医科大学学报》2025年第2期240-246,共7页Journal of Guizhou Medical University

基　　金：湖南省自然科学基金面上项目(2022JJ30845)。

摘　　要：目的探讨同源盒基因B7(homeobox gene B7,HOXB7)调控Wnt通路对骨肉瘤细胞上皮-间充质转化(epithelial-mesenchymal transition,EMT)及侵袭、迁移的影响。方法以体外培养的人骨肉瘤细胞株MG63、143B、SaoS2及人间充质干细胞hMSCs为研究对象,通过实时荧光定量PCR(qRT-PCR)和蛋白质印迹法(Western blot)测定HOXB7 mRNA和蛋白表达;将MG63细胞分为对照组、si-NC组、si-HOXB7组、si-HOXB7+SKL2001组及si-HOXB7+XAV939组,采用Transwell小室和划痕实验检测细胞侵袭和迁移,Western blot检测基质金属蛋白酶(matrix metallopeptidase,MMP)-2、MMP-9、Wnt、波形蛋白(Vimentin)、β-连环蛋白(β-catenin)、原癌基因c-myc、上皮钙黏蛋白(E-cadherin)、细胞周期蛋白D1(CyclinD1)及神经钙黏蛋白(N-cadherin)表达。结果骨肉瘤细胞系MG63、143B、SaoS2中HOXB7 mRNA和蛋白表达水平高于人间充质干细胞hMSCs(P<0.05),且HOXB7 mRNA和蛋白水平在MG63细胞中最高,故选择MG63细胞进行后续功能验证实验;沉默HOXB7表达后,HOXB7表达、侵袭和迁移细胞数、划痕愈合率及MMP-2、MMP-9、CyclinD1、Vimentin、N-cadherin、Wnt、核β-catenin及c-myc蛋白表达下降(P<0.05),E-cadherin、质β-catenin蛋白表达升高(P<0.05);SKL2001可逆转上述指标变化,而XAV939能促进这种变化(P<0.05)。结论沉默HOXB7可抑制骨肉瘤细胞侵袭、迁移和EMT,这与抑制Wnt通路相关蛋白表达有关。Objective To investigate the effect of homeobox gene B7(HOXB7)in regulating the Wnt signaling pathway on epithelial-mesenchymal transition(EMT),invasion,and migration in osteosarcoma cells.Methods Human osteosarcoma cell lines(MG63,143B,SaoS2)and human mesenchymal stem cells(hMSCs)were cultured in vitro.HOXB7 mRNA and protein expression were quantified using quantitative real-time PCR(qRT-PCR)and Western blot.MG63 cells were divided into five groups:control,small interfering negative control(si-NC),HOXB7-silenced(si-HOXB7),si-HOXB7+Wnt activator SKL2001,and si-HOXB7+Wnt inhibitor XAV939.Cell invasion and migration were assessed using Transwell chamber assays and wound healing assays.Western blot analysis measured the expression of matrix metallopeptidases(MMP-2,MMP-9),Wnt,Vimentin,β-catenin,c-myc,E-cadherin,CyclinD1,and nerve cadherin(N-cadherin).Results HOXB7 mRNA and protein expression levels were significantly higher in the osteosarcoma cell lines MG63,143B,and SaoS2 than in hMSCs(P<0.05),with the highest levels observed in MG63 cells.Thus,MG63 cells were selected for subsequent functional validation experiments.Silencing HOXB7 expression led to a decrease in HOXB7 expression,invasive and migratory cell numbers,wound healing rate,and the expression of MMP-2,MMP-9,Cyclin D1,vimentin,N-cadherin,Wnt,nuclearβ-catenin,and c-myc proteins(P<0.05),while the expression of E-cadherin and cytoplasmicβ-catenin proteins increased(P<0.05).SKL2001 reversed these changes,while XAV939 promoted them(P<0.05).Conclusion Silencing HOXB7 inhibits osteosarcoma cell invasion,migration,and EMT,which is associated with the downregulation of Wnt signaling pathway-related protein expression.

关 键 词：同源盒基因B7 WNT通路 骨肉瘤 侵袭 迁移 上皮-间充质转化 

分 类 号：R738.1[医药卫生—肿瘤]