Experimental Study Of The Effects Of Angiogenesis Inhibitor TNP-470 And TNP-470 Combined With BCNU On Mouse G422 Glioblastoma In Vivo

Objective: To investigate the feasibility of the treatment for the mouse G422 glioblastoma in vivo with angiogenesis inhibitor TNP-470 and TNP-470 combined with cytotoxic drug BCNU, and also analyze the mechanism of anti-angiogenesis and tumor-growth inhibiting of TNP-470.Methods: 72 adult female Kunming mouse were implanted subcutaneous of 1X107 (O.lml) G422 cells in the right leg. The animals were randomly divided into four groups 4 days after the implantation. In group 1, 4 days after the implantation , TNP-470 was administrated subcutanceously at the dose of 30mg/kg every other day for 8 times. In group 2 , BCNU was injected into peritoneal cavity at the dose of 20mg/kg at day 7> 9, 11 after the implantation and TNP-470 was administrated in the same manner as group 1. In the group 3 , BCNU was administrated in the same manner as group 2 . In group 4, the same volume of 3% ethanol ?5% arabic gum was administrated in the same manner as group 1 . The tumor size and the weight of mice were recorded . The mice were sacrificed by dislocating the neck at the day when the tumor size reached 500mm3 , the time of tumor-bearing was recorded. Theexpression of mVEGF|M mRNA was quantitatively analyzed by RT-PCR. The proliferation index in the tumor cells was evaluated by PCNA immunohistochemical stain and the apoptosis index in the tumor cells was evaluated by TUNEL stain. All the data were analyzed by the SPSS10. 0 for windows statistical software package.Results: The average time of tumor-bearing of the four groups were 25.2?.5d , 46.3 ?.2d , 43.1 ?.8d , 20.0?.7d . The differences in the time of tumor-bearing between every two groups were both significant ( PO.05 ). In the period of experiment , the activity of the mice treated by TNP-470 was normal. The average weight of mice of the TNP-470 group and the control group, at day 14 after the implantation, were 27.3+l.lg , 27.6?.4g . The difference in the weight between the two groups above was not significant ( P>0.05 ). The average weight of mice of TNP-470 combined with BCNU group and BCNU group, at day 34 after the implantation, were 27.9?.9g , 28.3?.0g . There was no significant difference in the weight between the two groups above (P>0.05 ). The results of quantitative analysis by RT-PCR showed the expression of mVEGFi6 56.0?.9%, and there was no significant difference in the proliferation index between the two groups above (P>0.05 ). The apoptosis index in the tumor cells in TNP-470 group and the control group were 5. 7?1.5%, 2.5 ?1.0%, and there was significant difference in the apoptosis index between the two groups above, (P<0.01).Conclusions: Angiogenesis inhibitor TNP-470 can inhibit the growth of mouse G422 glioblastoma in vivo, and TNP-470 probably inhibit the angiogenesis of G422 glioblastoma by inhibiting the expression of mVEGF mRNA of tumor cells, and TNP-470 remarkably inhibit the growth of G422 glioblastoma in vivo induced probably by hypoxia-mediated apoptosis of5tumor cells. TNP-470 combined with cytotoxic drug BCNU can enhance the effect of inhibition of the growth of mouse G422 glioblastoma in vivo. No obvious side effects of TNP-470 were detected.