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Study of Apoptosis in MCF-7 and Its Mechanism Caused by VEGF siRNA Transfected into It

Author: ZhangZuoZuo
Tutor: GeYinLin
School: Qingdao University
Course: Biochemistry and Molecular Biology
Keywords: RNAi MCF-7 SiRNA VEGF survivin
CLC: R346
Type: Master's thesis
Year: 2011
Downloads: 16
Quote: 0
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Abstract


Objective:To investigate the effect of small interfering RNA(siRNA) targeting vascular endothelial growth factor(VEGF) on apoptosis and apoptosis-related gene survivin when it reduced VEGF expression in human breast cancer MCF-7 cells.Methods:Small interfering RNA(siRNA) targeting VEGF was designed and synthesized in vitro,then transfected it into MCF-7 cells by using LipofectamineTM 2000, Control group、Lipofectamine group、scramble control group (SCR)and 3 siRNA concentration gradient groups (50、100、200nmol/L) have been set up.24 hours after transfection, apoptosis was examined by FCM essay.RT-PCR was used to examine the expression of survivin mRNA and Immunocytochemistry was used to evaluate the expression of survivin protein.Results:Compared with the control group,Small interfering RNA(siRNA) targeting VEGF after transfected into MCF-7 cells. the apoptosis of the 50nmol/L group was Increased (p<0.05),and the 100、200nmol/L groups was significantly increased (p<0.01).But the Lipofectamine group, the scramble control group (SCR) and the Control group were not statistically significant (p>0.05). The expression of survivin mRNA in all siRNA transfected groups were down-regulated,compared with the control group. (p<0.05, and 100nmol/L和200nmol/L groups p<0.01) the lipofectamine group、scramble control group (SCR), compared with the control group, were not statistically significant (p>0.05). survivin protein expression levels in the Down concentration of Transfected group, (p<0.05, and 100nmol/L和200nmol/L groups p <0.01), Relatively with the untreated group, the Lipofectamine group and the scramble control group (SCR) no significant difference(p>0.05).Conclusion:SiRNA targeting VEGF gene transfected in MCF-7 cells can up-regulation the apoptosis of MCF-7 cells, And its molecular mechanism was associated with the expression of survivin

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