Authors : Xin Fu, Weihua Li, Jian Zhang and YingHui Xu
Abstract: To study the role of P21/53 in directly regulating and controlling TIMP-2 and the important mechanism for promoting invasion of GSCs by P21/53. Transwell in vitro invasion experiment was used to test invasion ability of GSCs and GCs as well as effect of P21/53 on GSCs invasion. Techniques of qRT-PCR and Western blot were used to test and transfect expression variation of TIMP-2, MMP2 and MMP9 in GSCs of P21/53 inhibitor. The luciferase reporter gene experiment was used to test the role of P21/53 in directly regulating and controlling TIMP-2. The number of invasive GSCs significantly decreased following transfection of P21/53 inhibitor (p<0.05). The expression levels of TIMP-2 mRNA in the U87 cell line and GSCs from the two primary glioma specimens rose compared with the control group, respectively while the expression levels of MMP2 mRNA and MMP9 mRNA fell compared with the control group (p<0.05) following transfection of P21 inhibitor. Expression of protein level of TIMP-2, MMP2 and MMP9 were consistent with mRNA level and had significant difference compared with those of the control group following transfection of P21 inhibitor and P53 inhibitor. The luciferase activity of the reporter vector containing wiled-type TIMP-2-3'UTR increased significantly following transfection of P21/53 while the luciferase activity of the reporter decreased significantly following transfection of P21/53 simulator (p<0.05). High expression of P21/53 promotes invasiveness of GSCs and its mechanism is achieved by down-regulating expressions of TIMP-2 and up-regulating expressions of MMP2 and MMP9 as a minimum.
Xin Fu, Weihua Li, Jian Zhang and YingHui Xu, 2013. Study on Mechanism for P21/53 Promoting Invasion of Human Glioma Stem Cells by Inhibiting TIMP-2 Expression. Journal of Animal and Veterinary Advances, 12: 1220-1227.