磷脂酰肌醇蛋白多糖-3对肝癌细胞增殖和凋亡的影响

目的探讨shRNA干预磷脂酰肌醇蛋白多糖（GPC）-3基因转录对肝癌细胞增殖和凋亡的影响。方法将GPC-3-shRNA插入pGPU6／GFP／Neo质粒，转染人肝癌tlepG2细胞，以Western Blot和荧光定量PCR分别分析GPC-3蛋白和mRNA表达；以nTr法分析HepG2细胞增殖；以流式细胞术、Annexin—V—PE／7-AAD及细胞凋亡-DNA ladder等分析细胞周期及细胞凋亡率。结果shRNA1转染HepG2细胞，GPC-3mRNA沉默效率为89．3％，与GPC-3蛋白下调一致；以shRNA转染HepG2细胞，增殖抑制率为71．1％；细胞周期阻滞在G1期，细胞凋亡率达6...

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Published in临床肝胆病杂志 Vol. 29; no. 11; pp. 863 - 866
Main Author 邰伯军 姚敏 顾星 时运 蔚丹丹 陈洁 郑文杰 姚登福 陆少林
Format Journal Article
LanguageChinese
Published 南通大学附属医院,临床医学研究中心,江苏,南通,226001 2013
Subjects
基因沉默
癌
磷脂酰肌醇蛋白聚糖类
细胞凋亡
细胞增殖
肝细胞
细胞凋亡
基因沉默
癌,肝细胞
磷脂酰肌醇蛋白聚糖类
细胞增殖
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ISSN1001-5256
DOI10.3969/j.issn.1001-5256.2013.11.015

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Summary:目的探讨shRNA干预磷脂酰肌醇蛋白多糖（GPC）-3基因转录对肝癌细胞增殖和凋亡的影响。方法将GPC-3-shRNA插入pGPU6／GFP／Neo质粒，转染人肝癌tlepG2细胞，以Western Blot和荧光定量PCR分别分析GPC-3蛋白和mRNA表达；以nTr法分析HepG2细胞增殖；以流式细胞术、Annexin—V—PE／7-AAD及细胞凋亡-DNA ladder等分析细胞周期及细胞凋亡率。结果shRNA1转染HepG2细胞，GPC-3mRNA沉默效率为89．3％，与GPC-3蛋白下调一致；以shRNA转染HepG2细胞，增殖抑制率为71．1％；细胞周期阻滞在G1期，细胞凋亡率达65．6％。结论资料显示shRNA干预GPC-3基因转录，可显著抑制肝癌细胞增殖，促进肝癌细胞凋亡。
Bibliography:TAI Bojun, YAO Min, GU Xing, et al. （ Research Center of Clinical Medicine, Affiliated Hospital of Nantong University, Nantong 226001, China）
Objective To investigate the effects of silencing glypican -3 （GPC -3） gene transcription by shRNA on the proliferation and apoptosis of hepatoma cells. Methods GPC - 3 - shRNA was inserted into pGPU6/GFP/Neo vector, and HepG2 cells were transfected with the vector. The mRNA and protein expression of GPC -3 was measured by fluorescence quantitative PCR and Western blot; the proliferation of HepG2 cells was evaluated by MTT assay; the cell cycle and apoptosis of HepG2 cells were analyzed by flow cytometry, Annexin -V -PE/7 -AAD staining, and apoptotic DNA ladder kit. Results After the HepG2 cells were transfected with GPC -3 -shRNA, the GPC -3 mRNA silencing rate was up to 89.3%, in accordance with the down - regulation of GPC - 3 protein ; the proliferation inhibition rate of HepG2 cells was as high as 71.1% ; the HepG2 cells were arrested in GI phase, and the apoptosis
ISSN:1001-5256
DOI:10.3969/j.issn.1001-5256.2013.11.015