分化和DNA结合抑制因子1基因对乳腺癌细胞株MCF-7增殖活性及分泌血管内皮生长因子的影响  

Relationship between differentiation and DNA binding inhibition factor genel expression and cell proliferation, vascular endothelial growth factor secretion in MCF-7 breast cancer cell line

作　　者：王川[1] 洪天姿[1] 张捷[1] 郭雯珲[1] 傅芳萌[1] 卢辉山[1] 

机构地区：[1]福建医科大学附属协和医院乳腺外科,福州350001

出　　处：《中华实验外科杂志》2012年第5期841-843,F0003,共4页Chinese Journal of Experimental Surgery

摘　　要：目的观察分化和DNA结合抑制因子1（Idl）基因对人乳腺癌细胞增殖活性以及分泌血管内皮生长因子（VEGF）能力的影响。方法将正义、反义Idl基因真核表达载体以及空载体分别转染乳腺癌MCF-7细胞株，经潮霉素筛选获得稳定细胞株；逆转录-聚合酶链反应（RT—PCR）和免疫细胞化学染色检测IdlmRNA及Idl蛋白、VEGF蛋白表达；细胞计数、噻唑蓝（MTV）比色法及流式细胞仪检测细胞增殖活性及细胞周期；酶联免疫吸附试验（ELISA）检测细胞上清液中VEGF浓度。结果与空载体转染细胞比较，转染正义Idl载体的MCF-7细胞中IdlmRNA和Idl蛋白、VEGF蛋白表达水平上调，细胞生长加速（P〈0．05）；而反义Idl载体转染组细胞Idl、VEGF表达下降，细胞增殖速度降低（P〈0．05）。流式细胞检测结果显示，正义Idl转染组增殖期细胞（S＋G2＋M）比例为（48．45±2．97）％，空载体组为（32．40±0．49）％，反义Idl组为（23．08±1．56）％，各组间差异有统计学意义（P〈0．01）；3组细胞凋亡指数分别为（3．26±1．28）％、（7．42±1．04）％和（11．83±1．59）％，组间差异有统计学意义（P〈0．01）；正义Idl转染组细胞上清液中VEGF浓度较空载体组显著增高（P〈0．01），反义Idl组VEGF分泌量则较空载体组显著降低（P〈0．05）。结论Idl基因与乳腺癌MCF-7细胞增殖活性、细胞周期调控及VEGF分泌能力相关。Objective To study the influences of differentiation and DNA binding inhibition factor genel （ Idl ） gene expression on cell proliferation and vascular endothelial growth factor （VEGF） secretion in MCF-7 breast cancer cell line. Methods Positive or negative Idl eukaryotic expression vector （Idl-pcDNA3.1 ＋/Hygro, Idl-peDNA3.1-/Hygro）, as well as empty vector （peDNA3.1/Hygro）, was transfected into the MCF-7 cells, respectively. Hygromycin was used to select stabilized transfeeted cell lines. Reverse transcription-polymerase chain reaction （RT-PCR） and cell immunocytochemical staining were performed to detect the expression of Idl mRNA, Idl and VEGF proteins. Cell proliferation activity was analyzed by using cell counting, methyl thiazol tetrazolium （MTT） assay and flow eytometry （FCM）. The concentration of VEGF in the supernatant from cell cultures was tested by using enzyme-linked immu-nosorbent assays （ELISA）. Results In MCF-7 cells transfected with positive Idl eukaryotic expression vector （Idl-pcDNA3. 1 ＋/Hygro）, the Idl expression was up-regulated. Significantly higher cell proliferating activity was found in Idl-pcDNA3.1 ＋/Hygro group than in the empty vector transfected group （ P 〈 0. 05）. Negative Idl eukaryotic vector （Idl-pcDNA3.1-/Hygro） transfeetion inhibited the Idl expression and proliferating activity （ P 〈 0. 05 ）. FCM revealed that the proportion of proliferating cells （ S ＋ G2 ＋ M） was （48.45±2. 97 ） % in the positive Idl transfected group, （32.40±0.49 ） % in the empty vector trans- feeted group, and （23.08±1.56）% in the negative Idl transfected group. There were significant differ- ences among these three groups （P 〈 0. 01 ）. The apoptotic index in the positive Idl transfected group, the empty vector transfeeted group, and the negative Idl transfected group was （ 3.26±1.28 ） % , （ 7.42±1.04） % and （ 11.83±1.59） % , respectively （ P 〈0. 01 ）. Compared to that in the empty vector trans-feeted

关 键 词：乳腺肿瘤 基因表达 细胞增殖 血管内皮生长因子 

分 类 号：R737.9[医药卫生—肿瘤]