Systematic multiplex polymerase chain reaction and reverse transcription-polymerase chain reaction analyses of changes in copy number and expression of proto-oncogenes and tumor suppressor genes in cancer tissues and cell lines.
Systematic multiplex polymerase chain reaction and reverse transcription-polymerase chain reaction analyses of changes in copy number and expression of proto-oncogenes and tumor suppressor genes in cancer tissues and cell lines.
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系统多重聚合酶链反应和逆转录聚合酶链反应分析癌症组织和细胞系中原癌基因和抑癌基因的拷贝数和表达变化。
DOI：
10.1002/elps.200406090
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发表时间：
2004
期刊：
影响因子：
--
通讯作者：
Yamamoto,Fumiichiro
中科院分区：
文献类型：
--
作者：
Yamamoto,Miyako;Metoki,Rikiya;Yamamoto,Fumiichiro
Systematic multiplex reverse transcription‐polymerase chain reaction (SM RT‐PCR) is distinguishable from other multiplex RT‐PCR methods by (i) utilization of primers that amplify sequences that fall within a single exon of the genes, (ii) utilization of genomic DNA as a calibration standard, and (iii) optimized PCR conditions that allow amplification of bands of similar intensity using genomic DNA template. We previously developed the human experimental systems of 68 glycosyltransferase genes, 39 Hox genes, and 26 integrin subunit genes, and analyzed the expression of those genes in human adult tissues. Here we report the establishment of an SM RT‐PCR system of proto‐oncogenes and tumor suppressor genes and the analysis of gene expression in human cancer tissues and cell lines. We also demonstrate that the SM RT‐PCR system, which was developed for cDNA expression analysis, could also be used successfully for more exquisite analysis of copy number changes in genomic DNA. We observed a decrease in band intensity of HRAS, TP73, CDKN2A, and CDKN2B genes in most of the breast and prostate cancer cell lines examined. The decrease in copy number of HRAS proto‐oncogene leads us to suspect the presence of tumor suppressor genes in the vicinity of this gene on chromosome 11p15.5.
