SHC SH2域结合蛋白1对肝细胞癌细胞恶性生物学行为的影响及其作用机制  

Effects of SHC SH2 domain-binding protein 1 on the malignant biological behaviors of hepatocellular carcinoma cells and its mechanism

作　　者：宋磊 高明[2] 李贺[2] 孙远松 王琪 SONG Lei;GAO Ming;LI He;SUN Yuansong;WANG Qi(Emergency Department,the First Affiliated Hospital of Anhui Medical University,Anhui Hefei 230022,China;Department of Emergency Surgery,the Second Hospital of Anhui Medical University,Anhui Hefei 230022,China)

机构地区：[1]安徽医科大学第一附属医院急诊科,安徽合肥230022 [2]安徽医科大学第二附属医院急诊外科,安徽合肥230001

出　　处：《现代肿瘤医学》2023年第19期3548-3557,共10页Journal of Modern Oncology

基　　金：安徽省教育厅科学研究项目(编号:KJ2021A0322)。

摘　　要：目的:探究SHC SH2域结合蛋白1(SHCBP1)通过与驱动蛋白家族成员18A(KIF18A)相互作用对肝细胞癌(HCC)细胞增殖、凋亡、侵袭、迁移及干性特征的调控作用。方法:ENCORI数据库分析SHCBP1、KIF18A在HCC组织中的表达以及与HCC患者总体生存率之间的相关性以及SHCBP1与KIF18A表达在HCC组织中的相关性。分析SHCBP1、KIF18A与HCC患者临床病理特征间的关系;STRING数据库预测SHCBP1与KIF18A间潜在的结合关系。体外培养人正常肝细胞系HHL-5、HCC细胞(Huh-7、SNU-449、Hep3B),实时荧光定量PCR(RT-qPCR)、蛋白质印迹法(Western blot)检测SHCBP1、KIF18A表达水平;CCK-8检测细胞活力;TUNEL、划痕和Transwell实验分别评估细胞凋亡、迁移和侵袭;体外成球实验检测细胞成球能力;Western blot分析凋亡、转移相关蛋白及干性标志物表达水平。结果:SHCBP1、KIF18A表达均与HCC患者肿瘤分化程度、脉管侵犯及远处转移之间具有显著相关性(均P<0.05);SHCBP1和KIF18A在HCC组织和细胞中高表达(P<0.05),并与预后不良相关。敲低SHCBP1可抑制细胞增殖、迁移、侵袭和干性并促进细胞凋亡(均P<0.01)。此外,SHCBP1与KIF18A相互作用,KIF18A过表达可逆转SHCBP1干扰对HCC细胞增殖、凋亡、迁移、侵袭及干性的作用(均P<0.01)。结论:SHCBP1可通过与KIF18A相互作用进而促进HCC细胞恶性生物学行为。Objective:To figure out the regulatory role of SHC SH2 domain-binding protein 1(SHCBP1)in the proliferation,apoptosis,invasion,migration and stemness of hepatocellular carcinoma(HCC)cells via interacting with Kinesin family member-18A(KIF18A).Methods:ENCORI data base analyzed SHCBP1 and KIF18A expression in HCC tissues,the correlation of SHCBP1 and KIF18A expression with the overall survival rate of HCC patients and the correlation between SHCBP1 and KIF18A in HCC tissues.The relationship between SHCBP1,KIF18A and clinicopathological characteristics of HCC patients was analyzed.STRING database predicted the potential binding relationship between SHCBP1 and KIF18A.Human normal hepatocyte line HHL-5 and HCC cells(Huh-7,SNU-449 and Hep3B)were cultured in vitro.RT-qPCR and Western blot examined SHCBP1 and KIF18A expression.Cell viability was estimated by CCK-8.Cell apoptosis,migration as well as invasion were respectively measured by TUNEL,wound healing and transwell assays.In vitro sphere formation assay evaluated the sphere formation ability of cells.Western blot tested the expression of apoptosis-,metastasis-associated proteins and stemness markers.Results:There were significant correlations between SHCBP1 and KIF18A expression and the degree of tumor differentiation,vascular invasion and distant metastasis in HCC patients(all P<0.05).SHCBP1 and KIF18A were highly expressed in HCC tissues and cells(P<0.05)and associated with unfavorable prognosis.SHCBP1 knockdown hindered the proliferation,migration,invasion and stemness while potentiated the apoptosis of HCC cells(all P<0.01).Additionally,SHCBP1 interacted with KIF18A.Further,KIF18A over expression reversed the impacts of SHCBP1 interference on HCC cell proliferation,apoptosis,migration,invasion and stemness(all P<0.01).Conclusion:SHCBP1 might interact with KIF18A to further contribute to the malignant biological behaviors of HCC cells.

关 键 词：肝细胞癌 SHCBP1 KIF18A 细胞干性 

分 类 号：R735.7[医药卫生—肿瘤]