S期激酶相关蛋白1可抑制肝内胆管癌的增殖和侵袭

P. Yu, Q. Fu, Hao Zhang, Shu-Yi Zhao, Pan Liu, M. Hu
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The migration ability was detected by scratch test. The invasion ability of Transwell was detected by Transwellassay. \n \n \nResults \nThe expression of SKP1 in intrahepatic cholangiocarcinoma was (1.701±0.215) times, and the difference was statistically significant (t=8.512, P<0.05). 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=8.594, 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 Transwell 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 (t=9.918, P<0.05). \n \n \nConclusion \nThe expression of SKP1 in intrahepatic cholangiocarcinoma is higher than that in normal tissues. 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引用次数: 0

摘要

目的探讨s期激酶相关蛋白1 (SKP1)在肝内胆管癌中的表达及其对肝内胆管癌增殖、迁移和侵袭的影响。方法采用Western blotting法检测SKP1在肝内胆管癌组织与正常组织中的表达差异。构建SKP1干扰载体和空载体,通过慢病毒包装稳定转染肝内胆管癌细胞,形成实验组和对照组。免疫印迹法检测SKP1的表达。采用细胞计数试剂盒-8 (CCK-8)法检测转染后实验组和对照组细胞的增殖情况。通过划痕试验检测其迁移能力。采用Transwell法检测Transwell的侵袭能力。结果SKP1在肝内胆管癌组织中的表达量为(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SKP1表达降低,Western blotting法使实验组SKP1表达降低(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,实验组24、48、72 h的相对存活率分别为(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<0.01)。划痕试验显示,实验组在24、48小时的迁移细胞数(41.198±3.092)、(64.773±6.437)均显著低于对照组(113.487±3.514)、(168.767±8.151),差异有统计学意义(t=5.430、6.224,P<0.05)。Transwell实验显示,实验组穿孔细胞数为(34.185±3.124)个,显著低于对照组(121.234±6.814)个,差异有统计学意义(t=9.918, P<0.05)。结论SKP1在肝内胆管癌组织中的表达高于正常组织。降低SKP1表达可抑制肝内胆管癌的增殖、迁移和侵袭。关键词:S期激酶相关蛋白1;肝内胆管癌;泛素化;扩散;迁移;入侵
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S phase kinase-associated protein 1 can inhibit the proliferation and invasion of intrahepatic cholangiocarcinoma
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 Transwell was detected by Transwellassay. Results The expression of SKP1 in intrahepatic cholangiocarcinoma was (1.701±0.215) times, and the difference was statistically significant (t=8.512, P<0.05). 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=8.594, 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 Transwell 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 (t=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. Key words: S phase kinase associated protein-1; Intrahepatic cholangiocarcinoma; Ubiquitination; Proliferation; Migrate; Invasion
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