文档介绍：第三军医大学
硕士学位论文
反义核苷酸抑制GA表达对胃癌增殖能力影响的实验研究
姓名:刘孟刚
申请学位级别:硕士
专业:外科学(普外)
指导教师:顾红光
20040501
第三军医大学硕士学位论文
反义核苷酸抑制 GA 表达对胃癌增殖能力影响的实验研究

摘要

目的通过把转染了含谷氨酰***酶基因 C 末端反义序列质粒的胃癌细胞接种于裸
鼠观察其成瘤能力及增殖能力的变化检验反义核酸技术抑制谷氨酰***酶表达对肿
瘤生长的抑制作用为肿瘤的基因治疗提供新的理论依据
方法本课题前期阶段已将含谷氨酰***酶基因 C 末端反义序列的质粒转染入胃癌
细胞并命名为 SGC-7901GA 先把 SGC-7901GA细胞和空白对照组细胞 SGC-7901Neo 进
行体外培养再把两种胃癌细胞接种于 6 周龄 BALB/C 裸鼠皮下建立胃癌的裸鼠动
物模型观察裸鼠成瘤情况采用 MTT 法检测两种胃癌细胞在体外生长能力的差异
免疫组织化学方法(S-P 法)检测肿瘤组织增殖细胞核抗原(PCNA)表达 TUNEL 法检测
肿瘤组织细胞凋亡采用 RT-PCR 技术检测肿瘤组织中谷氨酰***酶 mRNA 的含量
结果
1. 两组细胞的生长曲线显示 SGC-7901GA 细胞增生速度明显低于对照组
2. 以相同的细胞浓度接种于裸鼠实验组成瘤时间明显晚于对照组成瘤后肿瘤
生长速度实验组也明显慢于对照组实验组肿瘤平均重量明显低于对照组
3. 实验组肿瘤组织的细胞增殖指数明显低于对照组而细胞凋亡指数明显高于对
照组
4. 肿瘤组织中谷氨酰***酶 mRNA 的含量实验组明显低于对照组
结论通过谷氨酰***酶基因 C 末端反义序列封闭谷氨酰***酶的基因抑制了谷氨
酰***酶的表达降低了胃癌细胞内谷氨酰***酶的活性阻断了胃癌对谷氨酰***的利用
从而抑制了肿瘤的增殖促进了肿瘤细胞的凋亡反义核苷酸技术抑制谷氨酰***酶表
达的方法是治疗肿瘤的一个新途径

关键词谷氨酰***谷氨酰***酶反义核苷酸胃癌肿瘤增殖细胞核抗原
细胞凋亡裸鼠 RT-PCR

5
第三军医大学硕士学位论文
An experimental study on the effect of using antisense
nucleotide technique to inhibit the expression of
gulutaminase gene

Abstract

Objectives: To estimate the inhibition effect on tumor growth after the glutaminase
gene was blocked with antisense nucleotide technique, we injected the gastric carcinoma
cells transfected with plasmid containing the C’ distal end antisense sequence of
glutaminase gene into nude mouse body pared its proliferation and neoplasia
capability with control gastric carcinoma provid a new academic basis for the gene
therapy of tumor.
Methods: The gastric carcinoma cells had been essfully transfected with plasmid
containing the C’ distal end antisense sequence of glutaminase gene and called
SGC-7901GA in previous researches. The SGC-7901GA cells and control cells SGC-7901Neo
were cultured in vitro,and then were respectively injected into the endermic tissue of
6-weeks-old BALB/C node mice to create the animal models of tumor. The neoplasia and
growth of tumor were observed. Use MTT method to