|
|
|
| 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 |
|
|
|
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.
|
|
Received: 23 December 2009
Published: 29 April 2010
|
|
|
| ［1］ Rana Tariq M. Illuminating the silence: understanding the structure and function of small RNAs. Nat Rev Mol Cell Biol,2007,8:2336.
［2］ McManus M T，Sharp P A．Gene silencing in mammals by small interfering RNAs．Nature,2002，3(10):737747.
［3］ Nykanen A, Haley B, Zamore P D. ATP requirements and small interfering RNA structure in the RNA interference pathway. Cell, 2001,107:309321.
［4］ Hannon Gregory J，Rossi John J．Unlocking the potential of the human genome with RNA interference.Nature,2004,431:371378.
［5］ Malsonpierre P C，Suri C．Jones P F，et at．Angiopoietin2.a natural antagonist for tie2 that disrupts in vivo angiogenesis. Science，1997,277(5322):5560.
［6］ George D Y，Samuel D，Nicholas W G，et al．Vascularspecific growth factors and blood vessel formation．Nature,20O0,407:242248.
［7］ Arai F，Hirao A，Ohmura M，et a1．Tie2／Angiopoietin1 signaling regulates hematopoietic stem cell quiescence in the bone marrow niche．Cell,2004,118:149161.
［8］ Fiedler U，Augustin H G．Angiopoietins：a link between angiogenesis and inflammation．Trends Immunol,2006,27(12):552558.
［9］ Jonathan Oliner，Hosung Min, Juan Leal, et al. Suppression of angiogenesis and tumor growth by selective inhibition of angiopoietin2. Cancer Cell,2004,6(5)：507516.
［10］ Rebekah R White，Siqing Shan，Christopher P, et al. Inhibition of rat corneal angiogenesis by a nucleaseresistant RNA aptamer specific for angiopoietin2. PNAS,2003,100:50285033.
［11］ Lin P，Buxton J A，Acheson A，et al．Antiagiogeni gene therapy targeting the endotheliumspecific receptor tyrosine kinase Tie2．Proc Natl Acad Sci USA,1998,95:88298834.
［12］ 王彪，单秀英，林菊丽，等. 促血管生成素2及其受体Tie2基因RNA干扰表达载体的构建与鉴定.医学综述,2009,15（21）：34983501. Wang B, Shan X Y, Lin J L, et al. Medical Recapitulate,2009,15（21）：34983501.
［13］ 林菊丽，王彪，黄祖根,等. 体外血管生成三维培养模型的构建. 福建医科大学学报，2009,43(01)4952 Lin J L, Wang B, Huang Z G, et al．Journal of Fujian Medical University，2009,43(01)4952.
［14］ Paddison P J, Hannon G J. siRNA and shRNAs: skeleton keys to the human genome. Curr Opin Mol Ther,2003,5(3):217224.
［15］ Zhang H, Fabrice A K, Lukasz J, et al. Single processing center models for human Dicer and bacterial RNase III.Cell, 2004,118: 5768.
［16］ Saharinen P, Kerkela K, Ekman N, et al.Multiple angiopoietin recombinant proteins activate the Tie1 receptor tyrosine kinase and promote its interaction with Tie2.J Cell Biol,2005,169:239240.
［17］ Xia X G, Zhou H, Ding H, et al. An enhanced U6 promoter for synthesis of short hairpin RNA. Nucleic Acids Res, 2003,31(17):100.
［18］ Dykxhoorn D M, Novina C D, Sharp P A. Killing the messenger: short RNAs that silence gene expression. Nat Rev Mol Cell Biol,2003,4(6):457467. |
|
Viewed |
|
|
|
Full text
|
|
|
|
|
Abstract
|
|
|
|
|
Cited |
|
|
|
|
| |
Shared |
|
|
|
|
| |
Discussed |
|
|
|
|
