Sugimoto T, Tsukamato F, Fujita M, Takai S
Department of Oncologic Surgery, School of Medicine, Osaka University, Japan.
Biochem Biophys Res Commun. 1994 Jun 15;201(2):574-80. doi: 10.1006/bbrc.1994.1740.
Ki-ras and c-myc oncogene mRNA in carcinoma tissue was quantitatively detected by the coamplification polymerase chain reaction. After the reverse transcription of the mRNA mixture with random hexanucleotide primer the coamplification polymerase chain reaction of the oncogene and the internal standard beta-actin cDNA was done. The amount of oncogene mRNA was calculated from the coamplified product ratio. Ki-ras expression in a human gastric cancer strain H-111 was estimated to 5 percent of beta-actin. The expression of c-myc mRNA in colorectal carcinoma tissue was observed to be ten to one hundred times higher than normal mucosa. Gene expression was compared numerically in mole/liter without using hybridization and radioactive probes.
采用共扩增聚合酶链反应定量检测癌组织中的Ki-ras和c-myc癌基因mRNA。用随机六核苷酸引物对mRNA混合物进行逆转录后，进行癌基因与内标β-肌动蛋白cDNA的共扩增聚合酶链反应。根据共扩增产物比例计算癌基因mRNA的量。估计人胃癌细胞系H-111中Ki-ras的表达量为β-肌动蛋白的5%。观察到结直肠癌组织中c-myc mRNA的表达比正常黏膜高10至100倍。无需使用杂交和放射性探针，以摩尔/升为单位对基因表达进行数值比较。
