INHIBITED NEOPLASTIC PHENOTYPE BY THE c-Ha-ras ANTISENSE RNA

A 2.0 kb fragment DNA plasmid which expresses antisense to the upstream first exon of c-Ha-ras oncogene was transfected into Ha-ras transformed cell lines, GCM-3T3 and REF-4.3. The transfection leads to the inhibition of malignant behaviour, shown by decreasing of growth speed, colony forming ability on soft agar, tumorigenicity in nude mice and increasing ot differentiation degree. In GCM-3T3 cells the lung metastasis frequency became much less (from 60% to 12.5%) after the transfection and the expression of ras oncogene product, and p21 protein was obviously decreased in the transfected cells. This work has first shown the inhibitory effect of antisense RNA on neoplastic behaviour in China.     

c-Ha-ras反义RNA对细胞恶性表型逆转作用的研究

本文应用表达反义c-Ha-ras癌基因上游区第一外显子2.0kb片段的RNA质粒,转染经c-Ha-ras转染的恶性细胞系GCM-3T3与REF-4.3,造成恶性行为的改变.表现在生长速度减慢,软琼脂克隆形成能力下降、裸鼠中致瘤力减小和肿瘤的病理分化程度增加,其中原来可造成转移的GCM-3T3细胞经反义RNA转染后肺转移率从60％降至12.5％,同时,两种细胞经转染后分离的克隆都有显著的ras癌基因产物p21蛋白表达的减少.本工作证明了反义RNA在癌基因表达上的调控和抑制细胞恶性行为的作用.