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KDR反义寡核苷酸对人胃癌细胞的作用
Effect of antisense oligodexynucelotide specific to KDR gene on human gastric carcinoma cell
【摘要】 目的 用 KDR反义寡核苷酸作用于人胃腺癌细胞 ,观察对细胞增殖及超微结构的影响 ,检测作用后 KDR基因的表达 ,并测定细胞分泌 VEGF的能力 .方法 MTT法测定细胞的增殖能力 ,电镜下观察细胞的超微结构 .原位杂交及免疫组化法检测 KDR m RNA及蛋白的表达 .ELISA法测细胞培养液中人 VEGF的含量 .结果 KDR ASODN可明显的抑制人胃腺癌细胞增殖 .增殖抑制率最高可达 55.4% .当剂量为 2 0μmol· L-1 时还可诱发细胞凋亡 .KDR ASODN能明显抑制细胞内 KDR m RNA和蛋白的表达 .胃癌细胞还能分泌一定量的人 VEGF:(92± 1 .7) ng· L-1 .结论 胃癌细胞能分泌一定量的人 VEGF.KDR ASODN能抑制细胞内 KDR基因的表达 ,并显著抑制胃癌细胞的增殖 .在一定剂量下还可诱发其凋亡 .说明 VEGF及受体 KDR在人胃腺癌细胞的增殖、凋亡的调控中起重要作用
【Abstract】 AIM VEGF and its receptor KDR play a key role in the tumor growth. To interrupt VEGF selfscrine way by using KDR ASODN acting on human gastric carcinoma cell (HGCC). To detecting the cell proliferation and expression of KDR gene. To observing the cellular ultrastructure and determining the VEGF level in the culture medium. METHODS Cell proliferation was determined by MTT. The ultrastructure was observed under electron microscope. VEGF level was determined by ELISA. KDR mRNA level was detected by in situ hybridization. The protein expression was detected by immunohistochemistry. RESULTS KDR ASODN can inhibit the proliferation of HGCC remarkably. The highest growth inhibitory rate is up to 55.4%. When its dose up to 20 μmol·L -1 , it can induce apoptosis. KDR ASODN can inhibit the expression of KDR mRNA and protein remarkably. HGCC can secret a certain level of VEGF: (92± 1.7) ng·L -1 . CONCLUSION VEGF and KDR play a key role in regulating the proliferation and apoptosis of HGCC. KDR ASODN can inhibit the proliferation of HGCC and reduce the expression of KDR gene remarkably. When up to a certain dose, it can induce apoptosis. HGCC can secret a certain level of VEGF. KDR ASODN may have the inhibitory effect on the tumor development.
- 【文献出处】 第四军医大学学报 ,Journal of The Fourth Military Medical University , 编辑部邮箱 ,2002年04期
- 【分类号】R735.2
- 【被引频次】15
- 【下载频次】59