Li Y, Fan M, Zhang J
Department of Biochemistry and Molecular Biology, China-Japan Friendship Institute of Clinical Medical Sciences, Beijing.
Zhonghua Zhong Liu Za Zhi. 1996 Jan;18(1):16-9.
The recombinant plasmid PGC was constructed for transcription unit of c-myc gene with bi-direction in vitro, to make RNA probes for detection of c-myc mRNA and antisense RNA expression of transfectant HLR60-9, which was obtained from HL60 cells transfected with inducible c-myc antisense RNA expression plasmid. The results from HLR60-9 cells induced by Cd2+ indicated that expression of c-myc antisense RNA increased with Cd2+ concentration and exposure time, while c-myc mRNA expression progressively reduced. Using immunohistochemical technique no c-myc P62 protein expression was detected. The incorporation of 3H-TdR,3H-UR,3H-Leu revealed significant suppression of DNA, RNA and protein biosyntheses. It is suggested that the reversion changes previously reported in malignant phenotypes of HLR60-9 cells and the inhibition of macromolecule biosynthesis mentioned above were associated with the blockade of c-myc gene expression by its antisense RNA.
构建重组质粒PGC用于体外双向转录c-myc基因单位,以制备RNA探针检测转染子HLR60-9中c-myc mRNA及反义RNA的表达,HLR60-9是由转染了诱导型c-myc反义RNA表达质粒的HL60细胞获得的。Cd2+诱导HLR60-9细胞的结果表明,c-myc反义RNA的表达随Cd2+浓度和暴露时间增加,而c-myc mRNA表达逐渐降低。采用免疫组化技术未检测到c-myc P62蛋白表达。3H-TdR、3H-UR、3H-Leu掺入显示DNA、RNA和蛋白质生物合成受到显著抑制。提示先前报道的HLR60-9细胞恶性表型的逆转变化及上述大分子生物合成的抑制与反义RNA阻断c-myc基因表达有关。