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Construction of Vectors with sFRP1-Fc、WIF1-Fc and Research the Role in Hepatoma

Author: HuangQingHong
Tutor: QianCheng
School: Zhejiang University of Technology
Course: Biochemistry and Molecular Biology
Keywords: Gene therapy CHO expression system ΦC31integrase Hepatocarcinoma
CLC: R735.7
Type: Master's thesis
Year: 2011
Downloads: 22
Quote: 0
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Abstract


Background :As a key of signaling pathways in the development of animal,Wnt signaling pathways involved in embryonic development from nematodes to humans and maintain adult normal structure of metabolism . It is an open signal pathway that it have lots of sites to be effected. This transduction pathway is extensive activation to regulate cell growth, migration and differentiation in lots of tumor of human. It has already been accepted as a tumor carcinogenesis of key signaling pathways. Thus it shows more and more potential influences in the stem cell biology and tumor and developmental biology. More and more research shows that any abnormal proteins would be concerned with the occurrence of cancer in recent years. Wnt signaling pathways involved the evolution .of breast, colon, cervical cancer, esophageal skin and other kinds cancer. Therefore for Wnt signaling pathways leading to the research and development on the signaling pathways and leading to the drug become a potentially treat cancer prospect direction. Diagnosis reagent and drug targeted Wnt signaling pathway is increasing currently.Objective:The aim of the experiment is to construct four new eukaryotic expression vector containing two inhibitory factor sFRP1- Fc and WIF1 -Fc which can inhibit the Wnt signaling pathways respectively. This kind of eukaryotic expression vector carries aφC31 integration sites attB which could be recombinated specially in the DNA sequences which contains attP or pseudo-attP sites to improve the rate of cloning formation .Before the plasmid polyclonal site there is a insulator sequence which can prevent gene expression reducding and instability of gene by integration. We explore that if the vector could increase the rate of formation of cloning in CHO and the enhance the expression of fusion protein.Therefore we detect if the expression vectors could restrain cancer cell proliferation and promote the rate of apoptosis In order to get a kind of new treatment for curding cancer.Methods: Constructing two eukaryotic expression vector contain fusion protein gene with Fc tag firstly. Then transfect two vectors into the CHO and screen the mono-colne then detect the expression quantity of two fusion protein. We treat normal Liver cells and liver cancer cells with vector to observe its function to human liver cell and conclude that expression of these vector expression capacity and tumor-kill capacity.Result: Construction of two eukaryotic vector which contains the sFRP1-Fc and WIF1-Fc respectively. And the experiment have shown that the vectors got a high expression of protein and theφC31 integrase could improve the rate of clone. The vectors containing sFRP1-Fc and WIF1-Fc could inhibit the proliferation of hepatoma cell.

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CLC: > Medicine, health > Oncology > Gastrointestinal Cancer > Liver tumors
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