细胞质内高表达的细胞周期蛋白依赖性激酶抑制蛋白P^(21)抑制人支气管上皮细胞凋亡  

Cytoplasmic highly-expressed cyclin-dependent kinase inhibitor P^(21) inhibits the apoptosis of human bronchial epithelial cell

作　　者：邹国明[1] 肖祖克[1] ZOU Guo-ming;XIAO Zu-ke(Department of Respiratory and Critical Medicine,Jiangxi Provincial People＇s Hospital,Nanchang 330006,Jiangxi,CHINa)

机构地区：[1]江西省人民医院呼吸与危重症医学科,江西南昌330006

出　　处：《海南医学》2018年第19期2665-2669,共5页Hainan Medical Journal

基　　金：江西省科学技术厅科技支撑计划(编号:2009BSA10700)

摘　　要：目的研究细胞周期蛋白依赖性激酶抑制蛋白P^(21)抑制人支气管上皮细胞凋亡的机制。方法 (1)表达细胞周期蛋白依赖性激酶抑制蛋白P^(21)的质粒P^(EGFP-N1-p21)转染人支气管上皮细胞系16HBE,研究P^(21)蛋白在细胞质、细胞核内的表达变化与质粒转染后细胞凋亡表现之间的相互关系。实验分组包括空白对照、空质粒及P^(EGFP-N1-p21)质粒组,应用Westen-blot方法检测质粒转染24 h后P^(21)蛋白在细胞质、细胞核内的表达变化相对值,流式细胞仪方法检测质粒转染24 h、48 h后的细胞凋亡率。(2)转化生长因子-β_1(TGF-β_1)刺激人支气管上皮细胞系16HBE,研究TGF-β_1刺激后P^(21)蛋白在细胞质、细胞核内的表达变化与支气管上皮细胞凋亡之间的相互关系。实验分组包括TGF-β_10 ng/mL、TGF-β_13 ng/mL及TGF-β_110 ng/mL组,应用Westen-blot方法检测TGF-β_1刺激24 h后P21蛋白在细胞质、细胞核内的表达变化,流式细胞仪方法检测TGF-β_1刺激12 h、24 h后的细胞凋亡率。结果质粒P^(EGFP-N1-p21)转染人支气管上皮细胞系16HBE 24 h后,P^(EGFP-N1-p21)质粒组的P^(21)蛋白在细胞质内表达水平高于空质粒组、空白对照组,并高于同组细胞核内,差异均有统计学意义(P<0.05);空质粒组、空白对照组的细胞质内P^(21)蛋白表达水平低于细胞核内,差异均有统计学意义(P<0.05)。P^(EGFP-N1-p21)质粒转染24 h、48 h后,P^(EGFP-N1-p21)质粒组凋亡率低于空质粒组及空白对照组,差异均有统计学意义(P<0.05)。P^(EGFP-N1-p21)质粒转染24 h后的16HBE细胞凋亡率高于转染48 h后,而空质粒组和空白对照组表现相反结果。P^(EGFP-N1-p21)质粒转染后,P^(21)蛋白在细胞质内表达与转染后的细胞凋亡呈负相关。TGF-β_1(0 ng/mL、3 ng/mL、10 ng/mL)作用于16HBE细胞24 h后,TGF-β_13 ng/mL及10 ng/mL组的细胞核、细胞质内P^(21)蛋白表达均高于TGF-β_10 ng/mL组,差异均具有统计学意义(P<0.05),TGF-β_13 ngObjectiveTo investigate the mechanisms of cyclin-dependent kinase inhibitor P21inhibiting the apoptosis of human bronchial epithelial cells（HBECs）.Methods The relationship of the expression of cytoplasm and nuclear cyclin-dependent kinase inhibitor P21with the apoptosis of 16HBE cells was studied after the plasmid PEGFP-N1-p21was transfected into the 16HBE cells,and the experiment included the blank,the empty plasmid and the PEGFP-N1-p21group.The expression of P21protein in cytoplasm and nucleus was detected by Western-blot after 24 hours of plasmid transfection.The apoptosis rate was detected by flow cytometry after 24 hours and 48 hours of plasmid transfection.The relationship of the expression of cytoplasmic and nuclear cyclin-dependent kinase inhibitor P21with the apoptosis of 16HBE cells was studied after the stimuli of transforming growth factor-β1（TGF-β1）to 16HBE cells,and the experiment included TGF-β10 ng/mL,TGF-β13 ng/mL,and TGF-β110 ng/mL group.The expression of P21protein in cytoplasm and nucleus was detected by Western-blot after 24 hours of the stimuli of TGF-β1.The apoptosis rate was detected by flow cytometry after 12 hours and 24 hours of the stimuli of TGF-β1.Results After the plasmid PEGFP-N1-p21was transfected into16HBE cells for 24 h,the cytoplasmic P21expression in the PEGFP-N1-p21group was higher than the nucleic P21expression,which was also significantly higher than the cytoplasmic P21expressions of the empty plasmid group and the blank group（P〈0.05）.Compared with the cytoplasmic P21expression,the nucleic P21expressions of the empty plasmid group and the blank group were higher（P〈0.05）.After the plasmid PEGFP-N1-p21was transfected for 24 hours and 48 hours,the apoptosis rate of 16HBE cells were significantly lower than the blank plasmid group and the control group（P〈0.05）.The apoptosis rate of 16HBE cells of 24 hours transfection in the plasmid PEGFP-N1-p21group was significantly higher than that of 48hours transfection（P〈0.05）,while the apoptosis rat

关 键 词：细胞周期蛋白依赖性激酶抑制蛋白P^21 转化生长因子-Β1 细胞凋亡 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]