Yamamoto Fumiichiro, Yamamoto Miyako
Cancer Genetics and Epigenetics Program, Burnham Institute for Medical Research, La Jolla, CA 92037, USA.
Electrophoresis. 2007 Jun;28(12):1882-95. doi: 10.1002/elps.200700093.
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.
我们运用自行研发的系统多重聚合酶链反应和逆转录聚合酶链反应（SM PCR和SM RT-PCR）方法，检测了乳腺癌和前列腺癌细胞系18q21-qter染色体区域上127个基因的拷贝数和表达差异。获得了半定量数据，其质量与DNA微阵列杂交分析的数据可比，但数量不可比。在乳腺癌、前列腺癌和其他癌症中频繁出现缺失的染色体区域，我们在MDA-MB-468乳腺癌细胞系中检测到SMAD4基因的纯合缺失。我们还观察到在某些乳腺癌和/或前列腺癌细胞系中，CCBE1、CCDC11、CD226、NP_115536.1、NP_689683.2、RNF152、SERPINB8和TCF4等基因出现部分或全部表达缺失。基因表达增加的情况罕见，但在所检测的所有癌细胞系中发现转录因子ONECUT2基因存在这种情况。实时定量逆转录聚合酶链反应实验证实了这些SM RT-PCR结果。通过实时定量逆转录聚合酶链反应对乳腺癌临床标本进行的进一步分析表明，在大多数乳腺癌临床病例中，CCBE1、TCF4、NP_115536.1和NP_689683.2的基因表达下调。
