FOXM1与CENP-A在骨肉瘤化疗耐药中的关系及其作用机制  

Relationship between CENP-A and FOXM1 in chemotherapy resistance of osteosarcoma and its mechanism

作　　者：黄宇航 尹诗源 李雨晴 尹玉[1] 蔡永萍[1,2] HUANG Yu-hang;YIN Shi-yuan;LI Yu-qing;YIN Yu;CAI Yong-ping(Department of Pathology,School of Basic Medical Science,Anhui Medical University,Hefei 230032,China;Department of Pathology,the First Affiliated Hospital of Anhui Medical University,Hefei 230022,China)

机构地区：[1]安徽医科大学基础医学院病理学教研室,合肥230032 [2]安徽医科大学第一附属医院病理科,合肥230022

出　　处：《临床与实验病理学杂志》2023年第4期445-450,共6页Chinese Journal of Clinical and Experimental Pathology

基　　金：安徽省高等学校自然科学研究重点项目(KJ2020A0142)。

摘　　要：目的探讨CENP-A是否参与骨肉瘤细胞化疗耐药及与叉头框蛋白M1(forkhead box M1,FOXM1)的调控关系。方法采用Western blot法检测骨肉瘤亲本细胞(HOS、U2OS)、顺铂耐药细胞(HOS/R、U2OS/R)和稳定FOXM1过表达细胞(HOS/FOXM1、U2OS/FOXM1)中CENP-A蛋白表达。转染siRNA后检测CENP-A蛋白表达;运用CCK-8细胞增殖实验观察siRNA敲低CENP-A后耐药骨肉瘤细胞对顺铂敏感性的影响;FOXM1抑制剂RCM1处理后检测CENP-A蛋白表达。Kaplan-Meier法分析GEPIA数据库中CENP-A表达与患者预后的关系,Spearman相关性分析CENP-A和FOXM1两者之间的关系。结果骨肉瘤顺铂耐药细胞中CENP-A蛋白表达比亲本细胞明显升高(0.52±0.03 vs 0.92±0.01,0.33±0.02 vs 1.12±0.01),FOXM1过表达细胞中CENP-A表达较空载体对照亦明显增高;在骨肉瘤顺铂耐药和FOXM1过表达细胞中转染siRNA-CENP-A后,CENP-A蛋白表达均明显降低(0.84±0.01 vs 0.60±0.02,0.98±0.01 vs 0.41±0.01)。转染siRNA敲低CENP-A后,骨肉瘤耐药细胞和FOXM1过表达细胞对顺铂的耐药性明显降低。10μmol/L RCM1 FOXM1抑制剂处理耐药细胞和FOXM1过表达细胞后,CENP-A蛋白表达均明显下降。生物信息学分析发现CENP-A高表达组患者总生存期显著低于低表达组(P<0.01)。Spearman相关性分析发现CENP-A和FOXM1表达呈正相关(r=0.79)。结论FOXM1可能通过调控CENP-A参与骨肉瘤化疗耐药,为骨肉瘤化疗耐药治疗策略提供新的理论基础。Purpose To investigate whether CENP-A is involved in chemotherapy resistance of osteosarcoma and its regulatory relationship with FOXM1.Methods The expression of CENP-A protein in osteosarcoma parental cells(HOS,U2OS),cisplatin-resistant cells(HOS/R,U2OS/R)and FOXM1 overexpressing cells(HOS/FOXM1,U2OS/FOXM1)was detected by Western blot.CENP-A protein expression was detected after siRNA knockdown of CENP-A.CCK-8 cell proliferation assay was used to observe the effect of siRNA knockdown of CENP-A on cisplatin sensitivity of drug-resistant osteosarcoma cells.The expression of CENP-A protein was detected after treatment with FOXM1 inhibitor RCM1.Kaplan-Meier method was used to analyze the relationship between CENP-A expression and patient prognosis in GEPIA database,and Spearman correlation analysis was used to analyze the correlation between CENP-A and FOXM1.Results The expression of CENP-A protein in cisplatin-resistant osteosarcoma cells was significantly higher than that in parental cells(0.52±0.03 vs 0.92±0.01,0.33±0.02 vs 1.12±0.01),and the expression of CENP-A in FOXM1 overexpressing cells was also significantly higher than that in empty vector control.The expression of CENP-A protein was significantly decreased after siRNA transfection in cisplatin-resistant(0.84±0.01 vs 0.60±0.02,0.98±0.01 vs 0.41±0.01)and FOXM1 overexpressing cells.After siRNA transfection to knock down CENP-A,the cisplatin resistance of drug-resistant osteosarcoma cells and FOXM1 overexpressing cells was significantly reduced.After treatment with 10μmol/L RCM1 FOXM1 inhibitor,CENP-A protein expression was significantly decreased in both drug-resistant cells and FOXM1 overexpressing cells.Bioinformatics analysis showed that the overall survival time of patients with high CENP-A expression was significantly lower than that of patients with low CENP-A expression(P<0.01).Spearman correlation analysis showed that CENP-A and FOXM1 expression were positively correlated(r=0.79).Conclusion FOXM1 may be involved in the chemotherapy resistan

关 键 词：骨肉瘤 化疗耐药 CENP-A FOXM1 顺铂 

分 类 号：R738.1[医药卫生—肿瘤]