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Expression of an intrabody against Human Cyclin Dl Provides Antitumor Efficacy on Hepatocellular Carcinoma both in vitro and in vivo

Expression of an Intrabody against Human Cyclin Dl Provides ...

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“细胞活动生命活力”——中国细胞生物学学会全体会员代表大会暨第十二次学术大会

作者： Bingzhi Li,Dan Yang,Hongliang Xu,Wenhan Wang,Yan Wu,Guiying Li Key Laboratory for Molecular Enzymology and Engineering of the Ministry of Education,Jilin University,Changchun,130021,China

Cyclin D1 is a rate-limiting key factor in controlling the transition from Gl to S in cell *** and overexpression of Cyclin D1 occurs in tumorigenesis of several types of human cancer including hepatocellular carcinoma(HCC).It becomes a potential target for the treatment of ***,by linking appropriate intracellular trafficking signal sequences to their coding genes,can be directed to all intracellular compartments to neutralize and block the function of target *** study was designed to use an intrabody against cyclin D1 to inhibit the growth and proliferation of HCC cells through blocking and inhibiting the biological activity of cyclin D1 overexpressed in cancer *** expression plasmid pER-ADK habouring an endoplasmic reticulum(ER)-retained scFv gene against human Cyclin D1(ER-ADκ) was *** pER-ADK was transfected into HepG2 *** RT-PCR and western blotting showed the anti-cyclinD1 intrabody was expressed efficiently in HepG2 *** assay showed that ER-ADk was localized in the endoplasmic *** results suggested that ER-ADk could recognize and combine its target protein cyclinD1 of HepG2 *** assay showed that ER-ADk significantly inhibited the growth of HepG2 cells(Pintrabody arrested most of HepG2 cells at G1 phase and distinctly induced apoptosis of HepG2 cells(Pintrabody inhibits the growth and proliferation of HCC cells both in vitro and in vivo by blocking cyclin D1 activity and it might be a potential approach for hepatocellular carcinoma therapy.

关键词： cyclin D1 intrabody HCC gene therapy

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The Role of the Anti-Cyclin D1 intrabody in the Hepatocellular Carcinoma Progression

The Role of the Anti-Cyclin D1 Intrabody in the Hepatocellul...

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第九届全国免疫学学术大会

作者： Yan Wu Xun Zhu An Cui Guannan Shen Hanwei Li Weiwei Tang Ying Zhang Ning Yang Guiying Li Key Laboratory for Molecular Enzymology and Engineering of the Ministry of Education Department of Immunology College of Bisic Medical SciencesJilin University

Background/Aims:Cyclin D1 plays an important role in the integration of mitogenic signals and promotion restriction point progression during G1 *** and overexpression of cyclin D1 occur in tumorigenesis of several types of human cancers,including hepatocellular carcinoma(HCC).Intrabodies can be directed to all intracellular compartments to neutralize and block the function of target *** study is designed to construct an intrabody against cyclin D1 and investigate its role in the hepatocellular carcinoma ***:A single-chain fragment of antibody variable region(scFv)against cyclin D1(AD1)was prepared by phage display ***,an expression plasmid plntra-AD1 habouring an endoplasmic reticulum(ER)-retained scFv gene against human cyclin D1(Intra-AD1)was *** expression and subcellular distribution of IntraAD1 in HepG2 cells were detected by RT-PCR,Western blot and fluorescence staining *** efficacy of Intra-AD1 on hepatocellular carcinoma in vivo and in vitro was *** mimic epitope peptides recognized by AD1 were obtained by phage peptides library display *** verify the results,the spatial structure of AD1 was modeled and docked with cyclin D1 by computer ***-immunoprecipitation analysis was used to investigate whether the AD1 affected the interaction between cyclin D1 and CDK4 or ***:The Intra-AD1 was expressed efficiently and could recognize and bind to cyclin D1 in HepG2/pintra-AD1 *** with the control,the expression of Intra-AD1 significantly inhibited the growth of HepG2 cells and tumors in vivo and in vitro,resulted in cell cycle arrest in the G0/G1 phase(p<0.01),and induced more cell apoptosis(p<0.01).Truncated mutation assay showed that the epitopes recognized by anti-cyclin D1 scFv(AD1)were in its N-terminal before amino acid *** this result was verified by computer modeling and *** the key amino acids recognized by

关键词： Cyclin D1 intrabody Hepatocellular carcinoma Protein interaction

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