shRNA干扰IRS-1基因通过PI3K/AKT通路对人乳头状甲状腺癌细胞TPC-1增殖和转移能力的调控作用  

Effects of interference IRS-1 with sh RNA on human papillary thyroid carcinoma cell line TPC-1 via PI3K/AKT signaling pathway

作　　者：江泽友[1] 徐灿[1] 葛一漫 JIANG Ze-You;XU Can;GE Yi-Man(Department of Clinical Laboratory,Affiliated Hospital of Chengdu University of Traditional Chinese Medicine,Chengdu 610072,China)

机构地区：[1]成都中医药大学附属医院检验科,成都610072

出　　处：《中国免疫学杂志》2018年第11期1674-1678,共5页Chinese Journal of Immunology

基　　金：四川省科技厅课题(2016SZ0037)

摘　　要：目的:探究下调胰岛素受体底物-1 (IRS-1)表达对人乳头状甲状腺癌细胞TPC-1增殖和转移能力的作用及作用机制。方法:将细胞分为TPC-1组,sh-scram组和sh-IRS-1组,用sh RNA IRS-1 (sh-IRS-1)转染sh-IRS-1组细胞,sh RNA转染sh-IRS-1组细胞,TPC-1组仅加入载体处理,RT-PCR和Western blot检测IRS-1的表达,MTT检测细胞增殖倍数,流式检测细胞凋亡,Transwell和划痕实验检测细胞侵袭、迁移能力,Western blot检测磷酸肌醇3激酶/蛋白激酶(PI3K/AKT)通路蛋白的表达。结果:与sh-scram组比较,sh-IRS-1组IRS-1的m RNA及蛋白表达水平明显降低; sh-IRS-1转染细胞后3 d和4 d,sh-IRS-1组细胞增殖倍数明显低于sh-scram组,细胞凋亡率明显高于sh-scram组;同时,与sh-scram组比较,sh-IRS-1组细胞侵袭及迁移能力显著降低;此外,sh-IRS-1还能显著降低p-PI3K/PI3K和p-AKT/AKT的比值。结论:干扰IRS-1表达能降低人乳头状甲状腺癌TPC-1细胞生存能力和转移能力,作用机制可能与抑制PI3K/AKT信号通路激活有关。Objective:To investigate effects and mechanisms of down-regulated insulin receptor substrate-1(IRS-1)on human papillary thyroid carcinoma(hPTC)cell line TPC-1.Methods:Cells were divided into TPC-1 group,sh-scram group and sh-IRS-1 group.Cells in sh-IRS-1 group were transferred with sh-IRS-1,cells in sh-scram group were transferred with shRNA,but cells in TPC-1 were treated with Lipofectamine 2000.The mRNA and protein levels of IRS-1 were detected by RT-PCR and Western blot.MTT assay was employed to measure growth rate and apoptosis rate was determined by flow cytometry.Invasion and migration abilities were measured by Transwell and wound healing assay.The expressions of phosphatidyl Inositol 3-kinase/AKT(PI3K/AKT)pathway-related proteins were measured by western blot.Results:Compared with sh-scram group,the mRNA and protein levels were decreased markedly.The cell growth rate was down-regulated and apoptosis rate was up-regulated compared with sh-scram group after cells were transferred with sh-IRS-1 for 3 d and 4 d.Meanwhile,compared with sh-scram group,the invasion and migration ability were decreased significantly.In addition,sh-IRS-1 decreased the ratio of p-PI3K/PI3K and p-AKT/AKT notably.Conclusion:Interference expression of IRS-1 inhibits cell growth and cell migration of hPTC cell line TPC-1,and the mechanism is related with inhibition of PI3K/AKT signaling pathway.

关 键 词：IRS-1 细胞增殖 细胞凋亡 侵袭 迁移 

分 类 号：R736.1[医药卫生—肿瘤]