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Ang2,Tie2 RNA Interference and the Function of Them on Inhibiting Angiogenesis in vitro |
WANG Biao1,LIU Zhao-liang1,LIN Ju-li1,SHAN Xiu-ying1,WANG Mei-shui1,ZHANG Sheng1,LU Kai-hua2 |
1.The First Affiliated Hospital, Fujian Medical University, Fuzhou 350005,China
2.The Centre of Plastic Surgery,The Fourth Military Medical University, Xi’an 710043,China |
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Abstract Objective: The expression of Ang2 and Tie2 is silenced by RNA interference in order to inhibit the angiogenesis in vitro. The study will supply the theory basis to the further animal research on the inhibition of tumor angiogenesis in vivo and provide the experimental evidence for tumor gene therapy in future. Methods: Using pSilencer 1.0-U6-Ang2/Tie2-siRNA recombinant plasmid to transfect into human umbilical vein endothelial cells (HUVECs), using RT-PCR to analyze the mRNA expression levels of Ang2 and Tie2 in HUVECs of each groups respectively. And the formation of vascular-like structure in HUVECs,transfected by the plasmids ,was studied in vitro by three-dimensional culture. Results: After the recombinant plasmids of pSilencer1.0-U6-Ang2/Tie2-siRNA transfected HUVECs,the mRNA expression levels of Ang2 、Tie2 in HUVECs were average obviously suppressed by RT-PCR(P<0.05),and there is no significant difference on the expression levels(P>0.05)between the two plasmids of each siRNA . There is a significant decrease on the quantity and length of vascular-like structure in HUVECs,transfected with the recombinant plasmids by three-dimensional culture model in vitro,(P<0.05).It proves that angiogenesis has been inhibited significantly in vitro. Conclusions:Ang2-siRNA、Tie2-siRNA can inhibit the mRNA expression of Ang2、Tie2 in HUVECs, and inhibit angiogenesis in the model of three-dimensional culture in vitro.
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Received: 23 December 2009
Published: 29 April 2010
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