Fang He-Qing, Li Hai-Jun, Liu Ying-Bin, Wang Xu-An, Ma Xiao-Ming, Kong Ying, Chen Yan, Chen De-Qin, Weng Wei-Hong, Zhang Zhi-Ping, Devkota K-R, Wang Jian-Wei, Li Jiang-Tao, Cao Li-Ping, Peng Shu-You
Department of Surgery, Second Affiliated Hospital, Medical School of Zhejiang University, Hangzhou 310009, China.
Zhonghua Yi Xue Za Zhi. 2007 Dec 18;87(47):3329-34.
To investigate the effects of vascular endothelial growth factor (VEGF) antisense oligodeoxynucleotide (ASODN) on the mRNA and protein expression of VEGF, Flt-1, and kinase insert domain containing receptor (KDR) and VEGF excretion in human gallbladder carcinoma cells.
Human gallbladder carcinoma cells of the line GBC-SD were cultured and transfected with VEGF ASODN and sense oligodeoxynucleotide (SODN) mediated by Oligofectamine. The toxicity of SODN and Oligofectamine to the GBC-SD cells was examined by MTT method. RT-PCR was used to detect the mRNA and expression of VEGF, Flt-1, and KDR, and ELISA was used to detect the protein expression of VEGF.
MTT method showed that SODN and Oligofectamine were not toxic to the GBC-SD cells. The mRNA expression levels of VEGF, Flt-1, and KDR of the ASODN and ASODN + Oligofectamine groups were all significantly lower than those of the control group (all P < 0.05), and were the lowest 72 hours after transfection, and then gradually increased. ELISA showed that there were not significant differences in the VEGF protein concentration in the supernatant of the GBC-SD cells among the SODN, SODN + Oligofectamine, and control groups (all P < 0.05), however, the VEGF protein concentration in the supernatant of the GBC-SD cells of the ASDN and ASDN + Oligofectamine groups were significantly lower than that of the control group (both P < 0.05).
VEGF ASODN inhibits the mRNA and protein expression of VEGF, Flt-1, and KDR and VEGF excretion in human gallbladder carcinoma cells. Oligofectamine strengthens the effect of ASODN.
探讨血管内皮生长因子（VEGF）反义寡脱氧核苷酸（ASODN）对人胆囊癌细胞中VEGF、Flt-1和含激酶插入结构域受体（KDR）的mRNA和蛋白表达以及VEGF分泌的影响。
培养人胆囊癌细胞系GBC-SD，并用脂质体介导的VEGF ASODN和正义寡脱氧核苷酸（SODN）进行转染。采用MTT法检测SODN和脂质体对GBC-SD细胞的毒性。应用RT-PCR检测VEGF、Flt-1和KDR的mRNA及表达情况，采用ELISA检测VEGF的蛋白表达。
MTT法显示SODN和脂质体对GBC-SD细胞无毒性。ASODN组和ASODN + 脂质体组的VEGF、Flt-1和KDR的mRNA表达水平均显著低于对照组（均P < 0.05），且在转染后72小时最低，随后逐渐升高。ELISA结果显示，SODN组、SODN + 脂质体组和对照组GBC-SD细胞培养上清液中的VEGF蛋白浓度差异无统计学意义（均P < 0.05），然而，ASDN组和ASDN + 脂质体组GBC-SD细胞培养上清液中的VEGF蛋白浓度显著低于对照组（均P < 0.05）。
VEGF ASODN可抑制人胆囊癌细胞中VEGF、Flt-1和KDR的mRNA和蛋白表达以及VEGF分泌。脂质体可增强ASODN的作用。
