多靶点蛋白酪氨酸激酶抑制剂Ponatinib对人肝癌细胞SK-Hep-1增殖、黏附和迁移能力的影响  

Effect of multi-target protein tyrosine kinase inhibitor Ponatinib on proliferation,adhesion and migration ability of human liver cancer cell line SK-Hep-1

作　　者：刘畅[1] 裴晋红[1] 穆秀丽 于保锋 LIU Chang;PEI Jinhong;MU Xiuli;YU Baofeng(Department of Biochemistry and Molecular Biology,Changzhi Medical College,Changzhi 046000,Shanxi Province,China;不详)

机构地区：[1]长治医学院基础部生物化学与分子生物学教研室,山西长治046000 [2]山西医科大学基础医学院生物化学与分子生物学教研室,山西太原030001

出　　处：《中国生物制品学杂志》2023年第6期668-672,679,共6页Chinese Journal of Biologicals

基　　金：山西省基础研究计划青年科学研究项目(20210302124183);山西省高等学校科技创新计划(2021L339);山西省自然科学基金基金项目:(201901D111190,2015011113);山西省重点研发计划(国际合作)(201703D421023);长治医学院博士科研启动基金(BS202007)。

摘　　要：目的探讨多靶点蛋白酪氨酸激酶抑制剂Ponatinib对人肝癌细胞SK-Hep-1增殖、黏附和迁移能力的影响。方法常规培养SK-Hep-1细胞,分别加入Ponatinib等24种酪氨酸激酶抑制剂,MTT法检测Ponatinib对SK-Hep-1细胞存活和增殖的影响。常规培养SK-Hep-1细胞至融合度达90%,分别加入0.1、0.5和1.0μmol/L Ponatinib,同时设对照组(不加Ponatinib),细胞缓慢聚集试验和细胞分离试验检测Ponatinib对SK-Hep-1细胞黏附能力的影响;划痕试验检测Ponatinib对SK-Hep-1细胞迁移能力的影响;Western blot法检测Ponatinib对SK-Hep-1细胞中E-cadherin蛋白表达水平的影响。结果24种酪氨酸激酶抑制剂对SK-Hep-1细胞均有抑制作用,其中Ponatinib抑制作用最强,IC_(50)达(0.288±0.044)μmol/L。与对照组比较,0.1、0.5和1.0μmol/L Ponatinib组细胞团块数明显减少(t分别为16.143、44.002、44.853,P均<0.001),含EDTA(TE)及CaCl_(2)(TC)胰酶消化后单个细胞数(N)的比值(N_(TC)/N_(TE))明显降低(t分别为4.276、10.625、27.571,P均<0.05),E-cadherin蛋白表达水平显著升高(t分别为-3.757、-4.561、-6.922,P均<0.05);划痕24及48 h后,0.1μmol/L Ponatinib组细胞划痕迁移率差异无统计学意义(t分别为0.473和0.872,P均>0.05),但0.5及1.0μmol/L Ponatinib组明显减小(t=6.272~16.733,P均<0.01)。结论Ponatinib可抑制SK-Hep-1细胞增殖和迁移,促进细胞黏附。Objective To investigate the effect of a multi-target protein tyrosine kinase inhibitor,Ponatinib,on proliferation,homogeneity adhesion and migration ability of human liver cancer cell line SK-Hep-1.Methods SK-Hep-1 cells were cultured routinely and added with 24 tyrosine kinase inhibitors such as Ponatinib respectively,and the effect of Ponatinib on the survival and proliferation of SK-Hep-1 cells was detected by MTT assay.SK-Hep-1 cells were cultured routinely until the fusion degree reached 90%,then added with 0.1,0.5 and 1.0μmol/L Ponatinib respectively,and the control group(without Ponatinib)was set up.The effect of Ponatinib on adhesion ability of SK-Hep-1 cells was detected by cell slow aggregation assay and dissociation assay,while the effect on migration ability by scratch test,and the effect on E-cadherin protein expression in SK-Hep-1 cells by Western blot.Results All 24 tyrosine kinase inhibitors inhibited SK-Hep-1 cells,among which Ponatinib showed the strongest inhibitory effect with a IC_(50) of(0.288±0.044)μmol/L.Compared with the control group,the number of cell mass(t=16.143,44.002 and 44.853 respectively,each P<0.001)and N_(TC)/N_(TE)[ratio of single cell number(N)after digestion by trypsin containing EDTA(TE)and CaCl_(2)(TC)](t=4.276,10.625 and 27.571 respectively,each P<0.05)decreased significantly and E-cadherin protein expression increased significantly(t=-3.757,-4.561 and-6.922 respectively,each P<0.05)in 0.1,0.5 and 1.0μmol/L Ponatinib groups;Scratch migration rate significantly decreased in 0.5 and 1.0μmol/LPonatinib groups(t=6.272~16.733 respectively,each P<0.01),while there was no significant difference in 0.1μmol/LPonatinib group(t=0.473 and 0.872 respectively,each P>0.05)after 24 h and 48 h of scratch.Conclusion Ponatinib inhibited proliferation and migration of SK-Hep-1 cells and promoted cell adhesion.

关 键 词：多靶点蛋白酪氨酸激酶抑制剂Ponatinib 肝癌 SK-Hep-1细胞 增殖 黏附 迁移 

分 类 号：R34[医药卫生—基础医学]