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Recombinant Plasmid pEGFP-AFP-hTNF-α Delivered by Nano-magnetic Fluids Killed Liver Cancer Cells (HepG2 Cells) in Vitro

Author: ZengJie
Tutor: PengJian
School: Central South University
Course: Surgery
Keywords: Nano - magnetic fluid AFP enhancer hTNFα gene Transfection Hepatic carcinoma Gene therapy
CLC: R735.7
Type: Master's thesis
Year: 2008
Downloads: 32
Quote: 0
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Abstract


Objective: To explore the nano- magnetic fluid as the feasibility of liver cancer gene therapy vectors . The 2 AFP enhancer ability to the foreign gene expression in AFP -positive cells within specific expression . Method : build AFP positive HepG2 cells enhanced sub- regulation containing the reporter gene green fluorescent protein , the target gene recombinant plasmid pEGFP-AFP-hTNFα hTNFα through nano - magnetic fluid were transfected into AFP expression and AFP negative expression of cervical cancer cell Hela. 12 hours after transfection , the dynamic observation under a fluorescence microscope for expression of the green fluorescent protein gene ; 48 hours after transfection , RT-PCR and Westernblot analysis of the the hTNFα the expression of the target gene in HepG2 cells . Taken to obtain the optimal transfection efficiency of DNA / gene transfection vector composite of the proportion of cell transfection, the transfection efficiency to compare different gene transfection vector CytometryAntibodies . HepG2 cell survival , MTT assay results were statistically processing . Apoptosis of HepG2 cells using flow cytometry . Results: Nano - magnetic fluid recombinant plasmid pEGFP-AFP-hTNFα can be transfected into HepG2 cells and transfection efficiency than liposomes ; hTNFα expression of the target gene transfected HepG2 cells by RT - PCR and Westernblot proof ; MTT and The flow cytometry Description hTNFα genetic to play killer cells role . Conclusion : 1 nanometer magnetic fluid capable of recombinant plasmid pEGFP-AFP-hTNFα transfected into HepG2 cells and expression as liver cancer gene therapy transfection vectors. 2 AFP enhancer enables directional rather specific expression of the target gene in HepG2 cells .

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