干预胰岛素样生长因子Ⅰ型受体活化联合抗癌药物抑制肝癌细胞增殖的协同作用分析

目的探讨干预胰岛素样生长因子Ⅰ型受体（IGF-ⅠR）基因转录,对抑制肝癌细胞增殖药物的协同作用。方法以p GPU6/GFP/Neo-IGF-ⅠR-shRNA高效质粒,转染IGF-ⅠR高表达的HBV阳性肝癌细胞PLC/PRF/5和HBV阴性Bel-7404,设空白对照组、阴性对照组和IGF-ⅠR-shRNA干预组;以荧光定量逆转录PCR和Western Blot分析RNA和蛋白表达;以细胞计数试剂盒分析细胞增殖;以流式细胞术、Annexin-V-PE/7-ADD分析细胞周期与凋亡。计量资料组间比较采用t检验,计数资料组间比较采用Fisher确切概率法。结果 IGF-ⅠR shRNA转染肝癌细胞P...

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Published in临床肝胆病杂志 Vol. 32; no. 8; pp. 1543 - 1548
Main Author 蔡胤 姚敏 王理 董志珍 顾娟娟 严晓娣 邱历伟 姚登福
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 2016
Subjects
受体,IGFⅠ型
奥沙利铂
癌,肝细胞
索拉非尼
索拉非尼
sorafenib
受体,IGFI型
奥沙利铂
receptor,IGF type 1
carcinoma,hepatocellular
癌,肝细胞
oxaliplation
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ISSN1001-5256
DOI10.3969/j.issn.1001-5256.2016.08.022

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Summary:目的探讨干预胰岛素样生长因子Ⅰ型受体（IGF-ⅠR）基因转录,对抑制肝癌细胞增殖药物的协同作用。方法以p GPU6/GFP/Neo-IGF-ⅠR-shRNA高效质粒,转染IGF-ⅠR高表达的HBV阳性肝癌细胞PLC/PRF/5和HBV阴性Bel-7404,设空白对照组、阴性对照组和IGF-ⅠR-shRNA干预组;以荧光定量逆转录PCR和Western Blot分析RNA和蛋白表达;以细胞计数试剂盒分析细胞增殖;以流式细胞术、Annexin-V-PE/7-ADD分析细胞周期与凋亡。计量资料组间比较采用t检验,计数资料组间比较采用Fisher确切概率法。结果 IGF-ⅠR shRNA转染肝癌细胞PLC/PRF/5效率为71%、肝癌细胞Bel-7404为90%,2种肝癌细胞IGF-ⅠR在mRNA和蛋白表达水平上同步减少;干预组细胞较阴性对照组均明显抑制,2组间肝癌细胞Bel-7404 72h时抑制率比较差异有统计学意义[（61.5±1.7）%vs（11.2±0.9）%,t=5.493,P〈0.05],2组间肝癌细胞PLC/PRF/5 72 h时抑制率比较差异有统计学意义[（63.9±3.9）%vs（9.5±1.1）%,t=19.244,P〈0.001]。干预组肝癌细胞Bel-7404凋亡率显著高于空白对照组（35.96%vs 12.16%,P〈0.001）和阴性对照组（35.96%vs 9.43%,P〈0.001）,干预组肝癌细胞PLC/PRF/5凋亡率显著高于空白对照组（44.84%vs 6.62%,P〈0.001）和阴性对照组（44.84%vs 4.02%,P〈0.001）;干预组肝癌细胞Bel-7404和PLC/PRF/5 G0/G1期百分率均明显高于阴性对照组[（59.0±1.3）%vs（48.4±0.8）%,t=12.032,P〈0.001;（65.4±0.5）%vs（53.5±0.7）%,t=22.789,P〈0.001）];干预组肝癌细胞Bel-7404和PLC/PRF/5周期蛋白cyclin D1表达均明显低于阴性对照组[（59.6±4.7）%vs（90.0±3.4）%,t=7.389,P〈0.05;（39.9±0.5）%vs（90.2±14.6）%,t=4.876,P〈0.05）];肝癌细胞Bel-7404和PLC/PRF/5干预组与阴性对照组在索拉非尼浓度分别为0、2.5、5、10、20 nmol/
Bibliography:carcinoma; hepatocellular; receptor; IGF type 1; sorafenib; oxaliplation
CAI Yin, YAO Min, WANG Li, et al. （ Research Center of Clinical Medicine, Affiliated Hospital of Nantong University, Nantong, Jiangsu 226001, China）
Objective To investigate the intervention of gene transcription of insulin- like growth factor- Ⅰ receptor（ IGF- ⅠR） and its synergistic effect with anti- cancer drugs in inhibiting the proliferation of hepatocellular carcinoma（ HCC） cells. Methods The HBV-positive HCC PLC / PRF /5 and HBV- negative Bel- 7404 cells were transfected with the efficient plasmid p GPU6 / GFP / Neo- IGF- ⅠR-shRNA. Fluorescent quantitative RT- PCR and Western blot were used to measure mRNA and protein expression,the Cell Counting Kit-8 was used to analyze cell proliferation,and flow cytometry and Annexin- V- PE /7- ADD were used to analyze cell cycle and apoptosis.The t- test was used for comparison of continuous data between groups,the Fisher＇s exact test was used for comparison of categorical data between groups.
ISSN:1001-5256
DOI:10.3969/j.issn.1001-5256.2016.08.022