S期激酶相关蛋白1与肝内胆管细胞癌细胞增殖、侵袭能力相关

S phase kinase-associated protein 1 can inhibit the proliferation and invasion of intrahepatic cholangiocarcinoma

目的观察人S期激酶相关蛋白1(SKP1)在肝内胆管细胞癌组织中表达差异并探讨其对肝内胆管细胞癌增殖、迁移及侵袭的影响.方法蛋白质印迹法(Western blot)实验测定SKP1在肝内胆管细胞癌癌组织与正常组织中表达差异;构建SKP1干扰载体及空载体,经慢病毒包装后稳定转染至肝内胆管癌细胞中,构成实验组与对照组,免疫印迹实验检测其SKP1表达变化;细胞计数试剂盒(CCK-8)实验检测转染后实验组与对照组增殖能力;划痕实验检测其迁移能力;Transwell实验检测其侵袭能力.结果肝内胆管细胞癌组织中SKP1表达为正常组织(1.701±0.215)倍,差异有统计学意义(t=8.512,P<0.05).慢病毒转染RBE细胞使其SKP1表达降低,蛋白质印迹法(Western blot)实验测定实验组较空白组降低(3.632±0.335)倍,差异有统计学意义(t=11.376,P<0.01).CCK-8实验显示24、48、72 h实验组较对照组相对生存率分别为(53.136±0.314)%、(42.661±0.248)%、(35.037±0.203)%,差异均有统计学意义(t=8.594、8.899、9.210,P<0.05).划痕实验显示24、48 h实验组迁移细胞数目[(41.198±3.092)、(64.773±6.437)个]较对照组[(113.487±3.514)、(168.767±8.151)个]明显降低,差异均有统计学意义(t=5.430、6.224,P<0.05).Transwell实验显示实验组穿孔细胞数目为(34.185±3.124)个较对照组[(121.234±6.814)个]明显降低,差异有统计学意义(t=9.918,P<0.05).结论肝内服管细胞癌组织中SKP1表达高于正常组织,降低SKP1表达能够抑制肝内胆管细胞癌的增殖、迁移及侵袭能力.

Objective To investigate the expression of S-phase kinase-associated protein 1(SKP1)in intrahepatic cholangiocarcinoma and to investigate its effects on proliferation,migration and invasion of intrahepatic cholangiocarcinoma.Methods Western blotting assay was used to determine the expression difference of SKP1 in intrahepatic cholangiocarcinoma and normal tissues.The SKP1 interference vector and empty vector were constructed and stably transfected into intrahepatic cholangiocarcinoma cells by lentivirus packaging to form an experimental group and a control group.The expression of SKP1 was detected by immunoblotting assay.The proliferation of experimental group and control group after transfection was detected by cell counting kit-8(CCK-8)assay.The migration ability was detected by scratch test.The invasion ability of Trans well was detected by Transwellassay.Results The expression of SKP1 in intrahepatic cholangiocarcinoma was(1.701±0.215)times,and the difference was statistically significant&512,PvO.05).Lentiviral transfection of RBE cells decreased the expression of SKP1,and the Western blotting assay decreased the experimental group by(3.632±0.335)times,and the difference was statistically significant(t=11.376,P<0.01).The CCK-8 experiment showed that the relative survival rates of the experimental group at 24,48,and 72 hours were(53.136±0.314)%,(42.661±0.248)%and(35.037±0.203)%,respectively,and the differences were statistically significant(t=6594,8.899,9.210,P<0.01).The scratch test showed that the number of migrated cells in the experimental group at 24 and 48 hours(41.198±3.092)and(64.773±6.437)were significantly lower than those in the control group(113.487±3.514)and(168.767±8.151),and the difference was statistically significant(t=5.430,6.224,P<0.05).The Trans well experiment showed that the number of perforated cells in the experimental group was(34.185±3.124),which was significantly lower than that in the control group(121.234±6.814),and the difference was statistically significant(i=9.918,P<0.05).Conclusion The expression of SKP1 in intrahepatic cholangiocarcinoma is higher than that in normal tissues.Decreasing SKP1 expression can inhibit the proliferation,migration and invasion of intrahepatic cholangiocarcinoma.