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Studies of Expression of Human VEGF Gene in Ovarian Cancer Cells with RNA Interference and Gene Therapy with Endostatin

Author: HanQingWang
Tutor: XuGenXing
School: Second Military Medical University
Course: Cell Biology
Keywords: RNA interference(RNAi) small hairpin RNA( shRNA) vascular endothelial growth factor(VEGF) ovarian cancer proliferation Bijidobacterium,shuttle vector endostatin
CLC: R737.31
Type: Master's thesis
Year: 2005
Downloads: 152
Quote: 0
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Abstract


RNA interference(RNAi) has been shown to interfere the expression of correlating mRNA in various species. This study proposed to inhibit the expression of vascular endothelial growth factor(VEGF) protein and the growth of cells in ovarian cancer cells. The vector which expresses the specific small hairpin RNA of VEGF gene was introduced into ovarian cancer HO-8910 cells by Lipofectamine 2000 and positive colones were screened by G418. The VEGF expression of HO-8910 cells before or after transfection were detected by Western blot, immunofiuorescent and RT-PCR methods, MTT and FCM.were used to observe the effect of proliferation to ovarian cancer cells. The results revealed that the expression of VEGF gene was inhibited in HO-8910-H1-VEGF cells by a small hairpin RNA of VEGF, the growth of ovarian cancer cells transfected with specific small hairpin RNA was slower, the G1 stage of the transfected cells was increased while the S stage was reduced. The specific small hairpin RNA targeting VEGF could inhibit the expression of VEGF protein and the proliferation of ovarian cancer cells.The research provided an experimental basis for cure human cancer with RNAi method. This study also constructed an Escherichia coli(E.coli) - Bifidobacterium longum(B. longum) shuttle expression vector, the expression products of the human endostatin gene was observed both in E. coli and B. longum. This study laid the base of using Bijidobacterium as a cancer gene therapy vector.

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CLC: > Medicine, health > Oncology > Genitourinary tumors > Female genital tumors > Ovarian tumors
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