Qiu Meng-Biao, Zhang Ji-Xiang, Liu Liang-Ming, Gong Bang-Dong, Wu Bo-Lin, Zhu Shui-Shan, Wen Yi
Molecular Center, Second Affiliated Hospital of Jiangxi Medical College, Nanchang 330006, China.
Hepatobiliary Pancreat Dis Int. 2004 Nov;3(4):552-7.
It is widely recognized that the growth of solid tumor depends on angiogenesis. Vascular endothelia growth factor (VEGF) is an endothelial cell-specific mitogen that promotes angiogenesis in solid tumor. Inhibition of angiogenesis is considered a promising approach for cancer therapy, and treatments including administration of antisense drugs and RNA interference for the VEGF gene are geared to the suppression of tumor angiogenesis.
As a new approach for gene therapy of hepatocellular carcinoma (HCC), four groups of antisense oligodeoxynucleotide (ASODN) (A-Cap, A-AUG, A-UGA and A-Exon-3) were used to block the expression of VEGF, then VEGF mRNA and protein were detected by RT-PCR and Western blot.
After treatment with ASODN, the relative VEGF mRNA levels of A-Cap, A-AUG, A-UGA, and A-Exon-3 were decreased significantly to (32+/-9)%, (63+/-1)%, (86+/-3)%, and (70+/-5)%, respectively(F=64.18, P<0.001). The relative VEGF protein levels of A-Cap, A-AUG, A-UGA and A-Exon-3 were decreased significantly to (41+/-5)%, (59+/-3)%, (88+/-7)%, and (79+/-9)% respectively (F=60.64, P<0.001).
Among the four ASODNs, the ASODN for Cap structure showed the strongest inhibitory effect and that for A-UGA, the least (P<0.05 ). The inhibitory effect of ASODN on the expression of VEGF proteins was similar to that of VEGF mRNA expression.
实体瘤的生长依赖于血管生成，这一点已得到广泛认可。血管内皮生长因子（VEGF）是一种内皮细胞特异性促有丝分裂原，可促进实体瘤中的血管生成。抑制血管生成被认为是一种有前景的癌症治疗方法，包括给予针对VEGF基因的反义药物和RNA干扰在内的治疗方法都旨在抑制肿瘤血管生成。
作为肝细胞癌（HCC）基因治疗的一种新方法，使用四组反义寡脱氧核苷酸（ASODN）（A-Cap、A-AUG、A-UGA和A-Exon-3）来阻断VEGF的表达，然后通过逆转录聚合酶链反应（RT-PCR）和蛋白质印迹法检测VEGF mRNA和蛋白质。
用ASODN处理后，A-Cap、A-AUG、A-UGA和A-Exon-3的相对VEGF mRNA水平分别显著降低至（32±9）%、（63±1）%、（86±3）%和（70±5）%（F = 64.18，P < 0.001）。A-Cap、A-AUG、A-UGA和A-Exon-3的相对VEGF蛋白水平分别显著降低至（41±5）%、（59±3）%、（88±7）%和（79±9）%（F = 60.64，P < 0.001）。
在这四种ASODN中，针对帽结构的ASODN抑制作用最强，针对A-UGA的ASODN抑制作用最弱（P < 0.05）。ASODN对VEGF蛋白表达的抑制作用与对VEGF mRNA表达的抑制作用相似。
