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背景与目的:舌癌是口腔常见的恶性肿瘤,目前常规采用以手术为主结合放疗化疗的综合治疗,总体的5年生存率只有50％左右,抗肿瘤血管生成治疗已 成为舌癌治疗的研究方向之一。本实验以5型E1缺陷型腺病毒携带的人内皮抑素基因(Ad/hEnd)感染舌癌细胞(Tca8113)和人脐静脉内皮细胞株(ECV),并对荷瘤裸鼠舌癌的抑瘤效果进行观察,研究其在舌癌细胞中的表达及对舌癌抑制作用。方法:(1)免疫组化法检测内皮抑素蛋白在Tca8113细胞和ECV细胞中的表达及分布。ELISA法检测上清中内皮抑素含量,Western blot检测内皮抑素基因在Tca8113和ECV细胞的表达特征。(2)流式细胞仪检测Ad/hEnd感染ECV后的细胞周期及凋亡,WST-1法检测Ad/hEnd对ECV细胞增殖的抑制。(3)Ad/hEnd对荷瘤裸鼠的舌癌的生长抑制分析。结果:(1)实验结果显示感染Ad/hEnd后Tca8113细胞和ECV细胞胞浆内可有效合成内皮抑素蛋白,细胞培养液上清中的内皮抑素蛋白表达浓度呈时间剂量依赖关系,最高达到597 ng/ml,可持续到第7天,并且表达产物有抑制人体静脉内皮细胞ECV生长特性,呈剂量依赖关系。(2)Ad/hEnd可延长感染后的ECV细胞的S期及G_2期,并出现细胞凋亡现象。(3)应用Ad/hEnd后第3天肿瘤体积增长受到抑制,第6天开始肿瘤抑制明显增强,第3周抑瘤率达45.8％。结论:本实?
BACKGROUND & OBJECTIVE: Tongue cancer is a common malignant tumor in oral cavity. At present, the comprehensive treatment of radiotherapy and chemotherapy combined with surgery is the routine method. The overall 5-year survival rate is only about 50%. Anti-tumor angiogenesis therapy has become a study of tongue cancer treatment One of the directions. In this study, tongue cancer cells (Tca8113) and human umbilical vein endothelial cell lines (ECV) were infected with human endostatin gene (Ad / hEnd) carried by type 5 E1-deficient adenovirus. Tumor effect was observed to study its expression in tongue cancer cells and tongue cancer inhibition. Methods: (1) The expression and distribution of endostatin protein in Tca8113 cells and ECV cells were detected by immunohistochemistry. The content of endostatin in the supernatant was detected by ELISA, and the expression of endostatin gene in Tca8113 and ECV cells was detected by Western blot. (2) The cell cycle and apoptosis after Ad / hEnd infection with ECV were detected by flow cytometry. The inhibition of Ad / hEnd on the proliferation of ECV cells was detected by WST-1 method. (3) Ad / hEnd growth inhibition of tongue cancer in nude mice. Results: (1) The results showed that endostatin protein was efficiently synthesized in the cytoplasm of Tca8113 cells and ECV cells infected with Ad / hEnd, and the concentration of endostatin protein in the cell culture supernatant was time-and dose-dependent up to 597 ng / ml, can be continued until the 7th day, and the expression product has inhibition ECV growth characteristics of human vein endothelial cells in a dose-dependent manner. (2) Ad / hEnd can prolong the S phase and G 2 phase of infected ECV cells and induce apoptosis. (3) On the third day after application of Ad / hEnd, the growth of tumor volume was inhibited. On the 6th day, the tumor inhibition was significantly enhanced. The inhibition rate of the 3rd week was 45.8%. Conclusion: The reality?