Study Of Mechanism Of Endostatin In The Treatment Of Gliomas

PARTⅠThe Expression and the Biological Significance ofβ-catenin and Cyclin D1 in GliomasObject:To invest the expression and the biological significance ofβ-catenin and Cyclin D1 in gliomas。Methods:The SABC immunohistochemistry methods were used to detect the expression ofβ-catenin and Cyclin D1 in 49 brain gliomas and 5 normal brain tissue。Result:Theβ-catenin positive ratio in normal brain tissues and glioma samples are 2/5 and 38/49(p<0.01),and the Cyclin D1 positive ratio in normal brain tissue are 1/5 and 42/49(p<0.01);Compared to the normal brain tissue, both the immunohistochemistric expression ofβ-catenin and Cyclin D1 were upgraded (p<0.05), and the immunohistochemistric expression ofβ-catenin and Cyclin D1 between the normal brain tissue and both groups of high grade and low grade glioma exhibited extremely significance statistics difference (p<0.01); correlation between the immunohistochemistry expression ofβ-catenin and Cyclin D1 shows statistics difference(r=0.6395,p<0.05)。Conclusion:β-catenin participated the development of gliomas, it activated the Cyclin D1, precipitate cell proliferation and tumorigenesis.PARTⅡThe study of the co-effect of Endostatin and Adriamycin to the proliferation and apoptosis in human glioma sarcomatosum cell line(GL15)Objects:To investigate the co-effect of Endostatin(ES) and combination of ES and adriamycin(ADM) to the proliferation and apoptosis in human glioma sarcomatosum cell line(GL15). Methods:Transfected the pcDNA 3.1 ES into GL15 cell, detected the effect of proliferation and apoptosis in GL15 cell by MTT and flow cytometer.Results:The inhibition of ADM to growth of the GL15 cell transfected pcDNA 3.1 ES was enhanced significantly, and proliferation was slowed significantly by MTT(P< 0.05).The flow cytometer detecte indicated that after transfected pcDNA 3.1 ES, the G0/G1 stage cell rate of GL15 cell line was increased significantly(P< 0.05), the decrease of the S stage cell rate and the upgrade of apoptosis were significantly(P< 0.01) too; Co-therapied by ADM, all the changes were more significantly(P< 0.01).Conclusion:The combination of ES and ADM could inhibit the growth of human glioma sarcomatosum cell line(GL15), their co-use could generate cooperational anti-tumor effect.PARTⅢEffect of endostatin on the expressiom ofβ-catenin and Cyclin D1 in GL15 glioma cellsObject: To investigate the effect of endostatin plasmid on the expressiom ofβ-catenin and Cyclin D1 in GL15 glioma cells.Methods: Transfected the pcDNA 3.1 ES plasmid into the GL15 glioma cells, and decteted the expression ofβ-catenin and Cyclin D1 protein in the transfected and not transfected GL15 glioma cells by SABC immunohistochemistry methods,and then analyse the difference beteen them.Result: The average ofβ-catenin and Cyclin D1 protein in the control group are 0.3758±0.0371 and 0.4092±0.0417,respectively;while the average ofβ-catenin and Cyclin D1 protein are 0.2462±0.0275 and 0.2046±0.0357 in the pcDNA 3.1 ES plasmid transfected group, respectively。The expression ofβ-catenin protein and Cyclin D1 protein are higher in the control groups than in the pcDNA 3.1 ES plasmid transfected groups, statistics analyze showed significantly statistics difference in the expression ofβ-catenin protein between the control group and the pcDNA 3.1 ES plasmid transfected groups (p<0.05), and the expression of Cyclin D1 protein between the control group and the pcDNA 3.1 ES plasmid transfected groups exhibited extremely significantly statistics difference (p<0.01).Conclusion: Endostatin could inhibit the overexpression and eptolexpression ofβ-catenin protein in the GL15 glioma cells,and inhibit the overexpression of Cyclin D1 protein in the GL15 glioma cells by direct and indirect pathway,and then inhibit the tumorigenesis.PartⅣThe study of co-therapy of endostatin and CTX in gliomas in vivoObject: To investigate the effect of co-therapy of endostatin and CTX in gliomas。Methods: Made the model of brain glioma by seeding cultuered the GL15 human glioma cell line into the subcutaneous of nude mouse; cured them by endostatin,CTX and both of them respectively, then object their effect。Result: Before study, the volumes of the subcutaneous tumor were 63.09±9.95mm3,61.42±6.07mm3,60.59±4.21mm3,62.22±7.10mm3, respectively; on the 6th day after therapy, the volumes of the subcutaneous tumor were 138.04±10.06mm3 ,108.61±8.57mm3,123.85±18.32mm3,103.26±8.32mm3,respectively ; on the 14th day after therapy, the volumes of the subcutaneous tumor were 450.12±73.23mm3 ,180.06±16.27mm3,322.69±99.40 mm3,154.91±16.79mm3;,respectively ; on the 22nd day after therapy, the volumes of the subcutaneous tumor were 2003.96±257.93mm3,176.61±30.13 mm3,844.67±110.44 mm3,146.4±8.30 mm3,respectively ; on the 28th day after therapy, the volumes of the subcutaneous tumor were 4267.96±313.59mm3 ,156.76±13.19mm3,1609.77±113.19mm3,134.52±10.38 mm3, respectively.Compared with the control group and the chemotherapy group, the endostatin group and the co-therapy group exhibited statistics difference since the 6th day after the treatment begining(p<0.05), exhibited extremely significance statistics difference since the 14th day(p<0.01), statistics difference appeared between the endostatin group and co-therapy group and between the control group and the chemotherapy group since 22nd day(p<0.05)。both the immunohistochemistry expression ofβ-catenin and Cyclin D1 exhibited extremely significance statistics difference (p<0.01) too; the immunohistochemistry expression betweenβ-catenin and Cyclin D1 shows statistics difference(p<0.05)。Conclusion: Endostatin could inhibit the growth of gliomas effectively,it could inhence CTX,seffect when they were used together。The mechanism is : at first ,endostatin inhibit the angiogenesis of glioma; secondly, endostatin regulate the expression ofβ-catenin and Cyclin D1 directly, and regulate the expression of Cyclin D1 indirectly, mediate the cell cycle of tumor cell,cause G1 arrest of GL15 cells.