FOXM1靶向HMMR对骨肉瘤细胞凋亡和自噬影响  

FOXM1 participates in apoptosis and autophagy of osteosarcoma cells by targeting HMMR

作　　者：翟东昌 唐磊 杨子杰 程明国[2] 张蓓[3] 沈若武[1] ZHAI Dongchang;TANG Lei;YANG Zijie;CHENG Mingguo;ZHANG Bei;SHEN Ruowu(Department of Special Medicine,School of Basic Medicine,Qingdao University,Qingdao 266071,China)

机构地区：[1]青岛大学基础医学院特种医学系,山东青岛266071 [2]青岛市第三人民医院骨科 [3]青岛大学基础医学院免疫学系

出　　处：《青岛大学学报（医学版）》2024年第2期261-266,共6页Journal of Qingdao University(Medical Sciences)

基　　金：青岛市自然科学基金项目(22-3-7-smjk-7-nsh)。

摘　　要：目的探讨叉头盒M1(FOXM1)转录因子通过靶向透明质酸介导的运动受体(HMMR)对骨肉瘤(OS)细胞凋亡和自噬的影响。方法使用慢病毒载体构建敲低FOXM1和过表达HMMR的OS细胞系,将转染空载慢病毒的细胞作为对照组,转染sh-FOXM1慢病毒的细胞作为敲低FOXM1组,转染sh-FOXM1慢病毒和OE-HMMR慢病毒的细胞作为过表达HMMR组。采用Western blot方法检测正常成骨细胞(hFOB1.19)和OS细胞(U-2OS)中FOXM1蛋白的表达;通过流式细胞术检测对照组、敲低FOXM1组和过表达HMMR组U-2OS细胞的凋亡率;采用Western blot方法检测对照组和敲低FOXM1组OS细胞中HMMR蛋白、自噬蛋白(LC3Ⅰ/Ⅱ)、凋亡相关蛋白(Cleaved-Caspase3、Bax)的表达以及过表达HMMR组OS细胞中自噬蛋白(LC3Ⅰ/Ⅱ)的表达。结果FOXM1在U-2OS细胞中的表达明显高于hFOB1.19细胞(t=38.780,P<0.005)。与对照组相比,敲低FOXM1组U-2OS细胞凋亡率显著增加(t=3.517,P<0.05),Cleaved-Caspase3、Bax、LC3Ⅰ/Ⅱ和HMMR蛋白表达显著下降(t=3.155~6.334,P<0.05),而过表达HMMR恢复了敲低FOXM1导致的凋亡率增加和自噬抑制(F=31.00、160.20,P<0.05)。结论FOXM1敲低通过降低HMMR蛋白的表达促进OS细胞凋亡和抑制OS细胞自噬。Objective To investigate the effects of forkhead box M1(FOXM1)transcription factor on apoptosis and autopha-gy of osteosarcoma(OS)cells by targeting hyaluronic acid-mediated motor receptor(HMMR).Methods The OS cell lines with FOXM1 knockdown and HMMR overexpression were constructed using the lentiviral vector.The cells transfected with the empty lentivirus were used as the control group,the cells transfected with the sh-FOXM1 lentivirus were used as the FOXM1 knockdown group,and the cells transfected with the sh-FOXM1 lentivirus and OE-HMMR lentivirus were used as the HMMR overexpression group.Western blot was used to detect the expression of FOXM1 in normal osteoblasts(hFOB1.19)and OS cells(U-2OS).The apoptosis rate of U-2OS cells in the control group,FOXM1 knockdown group,and HMMR overexpression group was determined by flow cytometry.Western blot was used to measure the expression of HMMR protein,autophagy protein(LC3Ⅰ/Ⅱ),and apoptosis-related proteins(Cleaved-Caspase3 and Bax)in the control group and FOXM1 knockdown group,as well as the expression of autophagy protein(LC3Ⅰ/Ⅱ)in the HMMR overexpression group.Results The expression of FOXM1 in U-2OS cells was significantly higher than that in the hFOB1.19 cells(t=38.780,P<0.005).Compared with the control group,the FOXM1 knockdown group showed significantly increased apoptosis rate of U-2OS cells(t=3.517,P<0.05),as well as significantly reduced expression of Cleaved-Caspase3,Bax,LC3Ⅰ/Ⅱ,and HMMR(t=3.155-6.334,P<0.05).Overexpression of HMMR restored the increase in apoptosis rate and inhibition of autophagy caused by FOXM1 knockdown(F=31.00,160.20;P<0.05).ConclusionFOXM1 knockdown promotes apoptosis and inhibits autophagy in OS cells by reducing HMMR protein expression.

关 键 词：叉头框蛋白M1 骨肉瘤 透明质酸受体 细胞凋亡 自噬 

分 类 号：R738.1[医药卫生—肿瘤] R329.25[医药卫生—临床医学]