Aurora激酶A抑制剂Alisertib对肝癌细胞DNA损伤修复的影响  

Effect of AURKA inhibitor Alisertib on DNA damage repair in hepatocellular carcinoma cells

作　　者：郝剑文 彭期臻 张雪宁[1] 秦宇[2] 李慧锴[3] HAO Jian-wen;PENG Qi-zhen;ZHANG Xue-ning;QIN Yu;LI Hui-kai(Department of Radiology,The Second Hospital of Tianjin Medical University,Tianjin 300211,China;Department of Diagnostics,College of Basic Medical Science,Tianjin Medical University,Tianjin 300070,China;Hepatobiliary Oncology,Tianjin Medical University Cancer Institute and Hospital,Tianjin 300060,China)

机构地区：[1]天津医科大学第二医院放射科,天津300211 [2]天津医科大学诊断学教研室,天津300070 [3]天津医科大学肿瘤医院肝胆肿瘤科,天津300060

出　　处：《中国临床药理学杂志》2020年第20期3257-3259,3263,共4页The Chinese Journal of Clinical Pharmacology

基　　金：国家自然科学基金资助项目(81502019);天津市自然科学基金资助项目(18JCYBJC93200)。

摘　　要：目的探讨Aurora激酶A(AURKA)抑制剂Alisertib对人肝癌细胞HepG2 DNA损伤修复的影响。方法将人肝癌细胞HepG2随机分为对照组(生理盐水)和低、中、高剂量实验组(0.01,0.1,1μmol·L^-1 Alisertib)。给药48 h后,用平板克隆形成法检测各组细胞克隆形成能力;用划痕愈合法检测各组细胞迁移侵袭能力;用Annexin V-FITC/PI双染法检测各组细胞凋亡情况;用蛋白质印迹法检测各组细胞AURKA及DNA损伤标记物p-H2A.X蛋白相对表达量。结果对照组,低、中、高剂量实验组克隆形成集落数分别为312.67±13.65,283.67±6.66,59.67±3.51,11.67±3.06;划痕面积百分比分别为(15.67±4.04)%,(34.00±5.00)%,(41.67±2.89)%,(76.00±2.65)%;总凋亡率分别为(1.34±0.29)%,(4.08±0.22)%,(7.26±0.93)%,(19.71±2.40)%;AURKA相对表达量分别为0.91±0.07,0.81±0.01,0.73±0.04,0.47±0.08;p-H2A.X相对表达量分别为0.26±0.05,0.52±0.07,0.89±0.08,1.12±0.18。低、中、高剂量实验组分别与对照组比较,差异均有统计学意义(均P<0.05)。结论Alisertib可通过下调AURKA蛋白表达,上调DNA损伤标记物p-H2A.X蛋白表达,有效激活DNA损伤修复,抑制肿瘤进展。Objective To investigate the effect of AURKA inhibitor Alisertib on DNA damage repair of human hepatocellular carcinoma cell line HepG2.Methods The human hepatocellular carcinoma cells HepG2 were randomly assigned to control group(normal saline)and test -L/-M/-H group(0.01,0.1,1μmol·L^-1 Alisertib).At 48 h after treatment,the clonality in each group were investigated by clone formation assay,the cellular invasion and migration ability in each group were determined by wound healing assay,the apoptosis in each group were detected by Annexin V-FITC/PI double staining,the expression of AURKA and DNA damage marker p-H2A.X of human hepatocellular carcinoma cells HepG2 were measured by Western blot.Results There were statistically significant differences between the control group and the test -L/-M/-H group in the number of colonies(312.67±13.65 vs 283.67±6.66,59.67±3.51,11.67±3.06),the scratch area rates[(15.67±4.04)%vs(34.00±5.00)%,(41.67±2.89)%,(76.00±2.65)%],the apoptotic rates[(1.34±0.29)%vs(4.08±0.22)%,(7.26±0.93)%,(19.71±2.40)%]and the relative expression of AURKA protein(0.91±0.07 vs 0.81±0.01,0.73±0.04,0.47±0.08),the relative expression of DNA damage marker p-H2A.X protein(0.26±0.05 vs 0.52±0.07,0.89±0.08,1.12±0.18)(P<0.05).Conclusion Alisertib can activate DNA damage repair of human hepatocellular carcinoma cell line HepG2 effectively through the down-regulation of AURKA protein and the up-regulation of DNA damage marker p-H2A.X,and inhibit the tumor progression.

关 键 词：肝细胞癌 Aurora激酶A Alisertib 细胞凋亡 DNA损伤 

分 类 号：R97[医药卫生—药品]