Identification of differentially expressed genes associated with HER-2/neu overexpression in human breast cancer cells
Identification of differentially expressed genes associated with HER-2/neu overexpression in human breast cancer cells
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DOI：
10.1093/nar/27.20.4008
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发表时间：
1999-10-15
影响因子：
14.9
通讯作者：
Slamon, DJ
中科院分区：
文献类型：
--
作者：
Oh, JJ;Grosshans, DR;Slamon, DJ
在类似于30%的人类乳腺癌和20%的人类卵巢癌中发现了HER-2/neu原癌基因的扩增和由此产生的过表达。为了更好地了解人类乳腺癌细胞中与该基因过表达相关的分子事件，采用差异杂交技术鉴定了在过表达该受体的细胞中表达水平发生改变的基因。从一个过表达细胞cDNA文库中筛选出16 000个克隆，共分离出19个非冗余克隆，其中7个与HER-2/neu过表达相关的表达降低（C克隆），12个与HER-2/neu过表达相关的表达升高（H克隆），其中5个C克隆和11个H克隆通过northern blot分析证实存在差异表达，其中9个基因功能已知。三个先前测序的功能相对不明确的基因和四个在GenBank中没有匹配的新基因。此外，在乳腺癌细胞系中鉴定出的16个（75%）cdna中，有12个在HER-2/neu过表达的卵巢癌细胞中也存在差异表达，这表明它们与HER-2/neu过表达具有普遍的相关性。最后，两种已鉴定的cdna，一种是新的，另一种是已鉴定但尚未表征的基因，在人类乳腺癌标本中被证实与HER-2/neu过表达有关。进一步表征这些基因可能有助于深入了解HER-2/neu过表达在人类乳腺癌和卵巢癌细胞中的基本生物学和发病作用。
Amplification and resulting overexpression of the HER-2/neu proto-oncogene is found in similar to 30% of human breast and 20% of human ovarian cancers. To better understand the molecular events associated with overexpression of this gene in human breast cancer cells, differential hybridization was used to identify genes whose expression levels are altered in cells overexpressing this receptor. Of 16 000 clones screened from an overexpression cell cDNA library, a total of 19 non-redundant clones were isolated including seven whose expression decreases (C clones) and 12 which increase (H clones) in association with HER-2/neu overexpression, Of these, five C clones and 11 H clones have been confirmed to be differentially expressed by northern blot analysis, This group includes nine genes of known function, three previously sequenced genes of relatively uncharacterized function and four novel genes without a match in GenBank. Examination of the previously characterized genes indicates that they represent sequences known to be frequently associated with the malignant phenotype, suggesting that the subtraction cloning strategy used identified appropriate target genes, In addition, differential expression of 12 of 16(75%) cDNAs identified in the breast cancer cell lines are also seen in HER-2/neu-overexpressing ovarian cancer cells, indicating that they represent generic associations with HER-2/neu overexpression, Finally, up-regulation of two of the identified cDNAs, one novel and one identified but as yet uncharacterized gene, was confirmed in human breast cancer specimens in association with HER-2/neu overexpression, Further characterization of these genes may yield insight into the fundamental biology and pathogenetic effects of HER-2/neu overexpression in human breast and ovarian cancer cells.