论文部分内容阅读
目的:探讨Endostatin基因(EScDNA)对大鼠胶质瘤的抑制作用。方法:采用脂质体法将pSecTagA-ES-cDNA和pSecTagA转染人脐静脉内皮细胞(EVC-304),并以 RT-PCR、Western Blot分别从RNA水平和蛋白水平鉴定转染是否成功。然后以 FCM测细胞凋亡情况,MTT实验测细胞活力,验证Endostatin基因对血管内皮细胞增殖的抑制作用。再以C_6胶质瘤细胞植于SD大鼠腋部皮下制作胶质瘤模型。将pSecTagA-EScDNA直接瘤内注射。分别观察记录肿瘤体积,绘制肿瘤生长曲线。结果:转染细胞PCR、Western分别检测到EScDNA和ES;FCM示细胞凋亡增加,MTT示细胞活力降低,肿瘤生长曲线示肿瘤生长明显受抑。说明Endostatin基因体内应用,具有明显抑制C_6胶质瘤的效应。结论:Endostin基因可能通过抑制肿瘤血管新生从而对胶质瘤达到抑制作用,为临床抗血管形成基因治疗胶质瘤提供了实验依据。
Objective: To investigate the inhibitory effect of Endostatin gene (ES cDNA) on glioma in rats. Methods: pSecTagA-ES-cDNA and pSecTagA were transfected into human umbilical vein endothelial cells (EVC-304) by liposome method. The transfection efficiency was evaluated by RT-PCR and Western Blot respectively. Cell apoptosis was measured by FCM and cell viability was measured by MTT assay to verify the inhibitory effect of Endostatin gene on the proliferation of vascular endothelial cells. C_6 glioma cells were transplanted subcutaneously in the axilla of SD rats to make a glioma model. The pSecTagA-ES cDNA was directly intratumorally injected. The tumor volume was observed and recorded, and the tumor growth curve was drawn. Results: EScDNA and ES were detected in transfected cells by PCR and Western blot, respectively. FCM showed that cell apoptosis increased, MTT showed cell viability decreased, and tumor growth curve showed significantly inhibited tumor growth. Endostatin gene in vivo application, with significant inhibition of C_6 glioma effect. Conclusion: Endostin gene may inhibit gliomas by inhibiting tumor angiogenesis and provide experimental evidence for clinical anti-angiogenic gene therapy of glioma.