Scanning copy number and gene expression on the 18q21-qter chromosomal region by the systematic multiplex PCR and reverse transcription-PCR methods.
We examined differences in copy number and expression of 127 genes located on the 18q21-qter chromosomal region of the breast and prostate cancer cell lines, using the systematic multiplex PCR and reverse transcription-PCR (SM PCR and SM RT-PCR) methods that we developed. Semi-quantitative data were obtained that were comparable in quality, but not in quantity, to data from DNA microarray hybridization analysis. In the chromosomal region where losses are frequent in breast, prostate, and other cancers, we detected a homozygous deletion of the SMAD4 gene in the MDA-MB-468 breast cancer cell line. We also observed partial or entire loss of expression in genes such as CCBE1, CCDC11, CD226, NP_115536.1, NP_689683.2, RNF152, SERPINB8, and TCF4 in certain breast and/or prostate cancer cell lines. An increase in gene expression was rare, but found with the transcription factor ONECUT2 gene in all of the cancer cell lines examined. Real-time qRT-PCR experiments confirmed these SM RT-PCR results. Further analysis of clinical specimens of breast cancer by real-time qRT-PCR demonstrated that the gene expression of CCBE1, TCF4, NP_115536.1, and NP_689683.2 was downregulated in the majority of clinical cases of breast cancer.
Yamamoto
Fumiichiro
F
Cancer Genetics and Epigenetics Program, Burnham Institute for Medical Research, La Jolla, CA 92037, USA. fyamamoto@burnham.org
Yamamoto
Miyako
M
D001943:Breast Neoplasms / Q000235:genetics* / Q000378:metabolism; D044584:Carcinoma, Ductal / Q000235:genetics* / Q000378:metabolism; D002460:Cell Line; D002887:Chromosomes, Human, Pair 18 / Q000235:genetics*; D017931:DNA Primers; D005260:Female; D005784:Gene Amplification / Q000235:genetics; D017353:Gene Deletion; D018628:Gene Dosage; D015870:Gene Expression; D020869:Gene Expression Profiling; D006801:Humans; D008297:Male; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods; D016133:Polymerase Chain Reaction / Q000379:methods; D011471:Prostatic Neoplasms / Q000235:genetics* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D014158:Transcription, Genetic; D014407:Tumor Cells, Cultured
D017931:DNA Primers