ShRNA干扰G蛋白偶联受体相关分选蛋白1对肺癌细胞增殖凋亡和裸鼠成瘤影响  

Effect of GASP-1-ShRNA on proliferation and apoptosis of lung cancer cells and tumor formation in nude mice

作　　者：韩贵良[1] 石岩[1] 焦婷[1] 贾友超[1] 任冠颖 HAN Gui-liang;SHI Yan;JIAO Ting(Hebei Province Key Laboratory for Cancer Radiotherapy and Chemotherapy,Department of Medical Oncology,Affiliated Hospital of Hebei University,Baoding,Hebei Province 071000,China)

机构地区：[1]河北大学附属医院肿瘤内科河北省放化疗机制与规程研究重点实验室,河北保定071000

出　　处：《中国公共卫生》2021年第6期978-981,共4页Chinese Journal of Public Health

基　　金：2019年保定市科技计划自筹经费(1941ZF084)。

摘　　要：目的探讨shRNA干扰G蛋白偶联受体相关分选蛋白1(GASP-1)对肺癌细胞增殖凋亡和裸鼠成瘤影响。方法将GASP-1-shRNA1、2和3用Lipofectamine 2000转染到肺癌A549细胞。选择转染GASP-1-shRNA1的细胞,建立空白对照组、空载体转染组和GASP-1-shRNA1转染组,采用菌落形成试验和流式细胞术分别检测肺癌A549细胞增殖和调亡;采用Western Blot检测ki67、cleaved caspase-3、Bax和Bcl-2蛋白表达量。将转染GASP-1-shRNA1细胞注射裸鼠后腿建立原位异种移植小鼠模型。检测肿瘤的生长,并通过免疫组化检测ki67和caspase-3蛋白表达量。结果GASP-1-shRNA1、2和3转染A549细胞后GASP-1 mRNA和蛋白表达量均显著下调,尤其转染GASP-1-shRNA1组。GASP-1下调能抑制A549细胞增殖,并促进A549细胞凋亡;在体内试验中,下调GASP-1能抑制体内肿瘤生长,同时也能抑制细胞增殖,并促进细胞凋亡。结论GASP-1的下调可抑制肺癌细胞增殖、促进凋亡,并抑制肿瘤生长。Objective To explore the effect of G-protein coupled receptor-associated sorting protein 1-short-hairpin RNA(GASP-1-ShRNA)on proliferation and apoptosis of lung cancer cells and tumor formation in nude mice.Methods GASP-1-shRNA1,GASP-1-shRNA2 and GASP-1-shRNA3 were transfected into lung cancer A549 cells with Lipofectamine 2000.Colony formation test and flow cytometry were used to detect proliferation and apoptosis of A549 cells from groups of GASP-1-shRNA1 transfection,blank control,and empty vector transfection,respectively.The A549 cells′expressions of Ki67,cleaved caspase-3,B cell lymphoma-2 protein(Bcl-2)and Bcl-2-associated X protein(Bax)were determined with Western blot.An orthotopic xenograft mouse model was established through injection of GASP-1-shRNA1-transfected cells into hind leg tissues in nude mice.The tumor growth of the model mice were observed and the expressions of ki67 and caspase-3 protein in tumor tissues were detected with immunohistochemistry.Results The mRNA and protein expression of GASP-1 in A549 cells were down-regulated after transfections of GASP-1-shRNA1,GASP-1-shRNA2,and GASP-1-shRNA3,especially in the GASP-1-shRNA1 transfected A549 cells.The down-regulation of GASP-1 could inhibit proliferation and promote cell apoptosis of the A549 cells.In vivo experiments,down-regulated GASP-1 could restrain tumor growth by inhibiting proliferation and promoting apoptosis of the A549 cells.Conclusion Down-regulation of GASP-1 could inhibit proliferation and promote apoptosis of lung cancer A549 cells.

关 键 词：shRNA干扰G蛋白偶联受体相关分选蛋白1(GASP-1) 肺癌 增殖 凋亡 

分 类 号：R99[医药卫生—毒理学]