>
网站首页期刊介绍通知公告编 委 会投稿须知电子期刊广告合作联系我们在线留言
最新消息:
人TRB3过表达载体的构建及检测
作者:李杨1 2  胡斌2  关亚萍2  徐敏1 2  孟祥军2 3 
单位:1. 南京医科大学上海一院临床医学院 消化科, 上海 200080;
2. 上海交通大学附属第一人民医院 消化科, 上海 200080;
3. 上海交通大学附属第九人民医院 消化科, 上海 200030
关键词:TRB3 过表达 肝细胞癌 
分类号:R734.7
出版年·卷·期(页码):2017·36·第一期(17-20)
摘要:

目的:构建TRB3真核表达重组质粒,并用肝癌细胞MHCC-97H检测其表达。方法:从肝癌组织中提取并扩增TRB3基因的编码区,将其克隆到真核表达载体pcdh-CMV-MCS-GFP中,酶切并检定,将克隆成功的质粒转染至MHCC-97H细胞中,用qPCR和Western Blot检测TRB3的表达水平。结果与结论:成功扩增了TRB3的编码区,并克隆至真核表达载体中;转染后72、96h,MHCC-97H细胞的TRB3 mRNA和蛋白表达显著增高;成功构建的TRB3过表达载体可用于后续实验。

Objective:To construct human TRB3 gene overexpression vector and identify its expression in hepatocellular carcinoma cells MHCC-97H.Methods:TRB3 gene was extracted from liver cancer tissue of human and amplified it by reverse transcription.Then it was cloned into pcdh-CMV-MCS-GFP vector for constructing eukaryotic expression vector.The successful cloned vector was transfected into hepatocellular carcinoma cells MHCC-97H,and TRB3 expression was determined by qPCR and Western blot.Results and Conclusion:The coding region of human TRB3 gene is successfully amplified,and cloned into the vector;the expression of TRB3 mRNA and protein are upregulated in MHCC-97H cells after transfected;TRB3 overexpression vector is successfully constructed,which could be used into the post-study.

参考文献:

[1] TORRE L A,BRAY F,SIEGEL R L,et al.Global cancer statistics,2012[J].CA Cancer J Clin,2015,65(2):87-108.
[2] 田贵金,刘军红,刘涛,等.白介素2联合胸腺肽α1对肝癌术后患者肝纤维化指标和肝功能的影响[J].现代医学,2016,35(1):68-71.
[3] GROSSHANS J,WIESCHAUS E.A genetic link between morphogenesis and cell division during formation of the ventral furrow in Drosophila[J].Cell,2000,101(5):523-531.
[4] DU K,HERZIG S,KULKARNI R N,et al.TRB3:a tribbles homolog that inhibits Akt/PKB activation by insulin in liver[J].Science,2003,300(5625):1574-1577.
[5] HEGEDUS Z,CZIBULA A,KISS-TOTH E.Tribbles:A family of kinase-like proteins with potent signalling regulatory function[J].Cell Signal,2007,19(2):238-250.
[6] XU J,LV S,QIN Y,et al.TRB3 interacts with CtIP and is overexpressed in certain cancers[J].Biochim Biophys Acta,2007,1770(2):273-278.
[7] 冯作化,药立波,方定志,等.生物化学与分子生物学[M].3版.北京:人民卫生出版社,2015:500-530.
[8] HAMID A S,LI J,WANG Y,et al.Recombinant human decorin upregulates p57KIP2 expression in HepG2 hepatoma cell lines[J].Mol Med Rep,2013,8(2):511-516.
[9] ZHANG J,WEN H J,GUO Z M,et al.TRB3 overexpression due to endoplasmic reticulum stress inhibits AKT kinase activation of tongue squamous cell carcinoma[J].Oral Oncol,2011,47(10):934-939.
[10] MIYOSHI N,ISHII H,MIMORI K,et al.Abnormal expression of TRIB3 in colorectal cancer:a novel marker for prognosis[J].Br J Cancer,2009,101(10):1664-1670.
[11] ERAZO T,LORENTE M,LOPEZ-PLANA A,et al.The new antitumor drug ABTL0812 inhibits the Akt/mTORC1 axis by upregulating Tribbles-3 pseudokinase[J].Clin Cancer Res,2016,22(10):2508-2519.
[12] SANO R,REED J C.ER stress-induced cell death mechanisms[J].Biochim Biophys Acta,2013,1833(12):3460-3470.
[13] BROMATI C R,LELLIS-SANTOS C,YAMANAKA T S,et al.UPR induces transient burst of apoptosis in islets of early lactating rats through reduced AKT phosphorylation via ATF4/CHOP stimulation of TRB3 expression[J].Am J Physiol Regul Integr Comp Physiol,2011,300(1):R92-100.
[14] LI T,SU L,ZHONG N,et al.Salinomycin induces cell death with autophagy through activation of endoplasmic reticulum stress in human cancer cells[J].Autophagy,2013,9(7):1057-1068.

服务与反馈:
文章下载】【发表评论】【查看评论】【加入收藏
提示:您还未登录,请登录!点此登录
您是第 124164 位访问者

copyright ©《东南大学学报(医学版)》编辑部
联系电话:025-87232483 83272481
电子邮件: bjb@pub.seu.edu.cn