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.
作者
韩贵良
石岩
焦婷
贾友超
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)
出处
《中国公共卫生》
CSCD
北大核心
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)。
