Differential screening and suppression subtractive hybridization identified genes differentially expressed in an estrogen receptor-positive breast carcinoma cell line
Differential screening and suppression subtractive hybridization identified genes differentially expressed in an estrogen receptor-positive breast carcinoma cell line
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DOI：
10.1093/nar/26.4.1116
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发表时间：
1998-02-15
影响因子：
14.9
通讯作者：
Weigel, RJ
中科院分区：
文献类型：
--
作者：
Kuang, WW;Thompson, DA;Weigel, RJ
基因表达的差异可能解释激素反应性乳腺癌和激素无反应性乳腺癌之间的表型差异。我们已经鉴定出雌激素受体（ER）阳性MCF7乳腺癌细胞系与ER阴性MDA-MB-231乳腺癌细胞系相比差异表达的cDNA。差异筛选分离出4个差异表达基因：细胞角蛋白8、细胞角蛋白18、Hsp27和GPCR-Br。为了鉴定丰度较低的差异表达基因，利用抑制消减杂交分离出29个差异表达克隆。序列分析显示，11 个克隆来自先前描述的基因：HEK8、神经肽 Y 受体 Y1、p21(WAF-1)、p55(PIK)、细胞角蛋白 18（克隆两次）、果糖 1,6-二磷酸酶、细胞角蛋白 8、TGF β 1 结合蛋白、延伸因子 1 α 2 和 pS2。其余18个克隆与GenBank/EMBL数据库中的序列不匹配，表明它们可能是新基因。 pS2、神经肽 Y 受体 Y1 和三个新克隆的表达由雌二醇诱导，表明雌激素反应性。一个新基因 DEME-6 的表达模式与一组乳腺癌细胞系中 ER 和 ERF-1/AP-2 γ 的表达相关。 DEME-6 的 2.6 kb cDNA 经测序，包含 574 个氨基酸的开放阅读框，与秀丽隐杆线虫 III 号染色体的基因有 62.4% 的相似性。 RT-PCR 测定，在原发性乳腺癌中也检测到 DEME-6 的表达，但在正常乳腺组织中未检测到 DEME-6 的表达。这些发现支持这样的假设：一组与 ER 协调调节但不一定与雌二醇反应的基因是激素反应性乳腺癌表型的特征。
Differences in gene expression are likely to explain the phenotypic differences between hormone-responsive and hormone-unresponsive breast cancer. We have identified differentially expressed cDNAs in the estrogen receptor (ER)-positive MCF7 breast carcinoma cell line compared with the ER-negative MDA-MB-231 breast carcinoma cell line. Differential screening isolated four differentially expressed genes: cytokeratin 8, cytokeratin 18, Hsp27 and GPCR-Br. To identify differentially expressed genes of lower abundance, suppression subtractive hybridization was utilized and 29 differentially expressed clones were isolated. Sequence analysis revealed that 11 clones were from previously described genes: HEK8, neuropeptide Y receptor Y1, p21(WAF-1), p55(PIK), cytokeratin 18 (cloned twice), fructose-1,6-biphosphatase, cytokeratin 8, TGF beta 1 binding protein, elongation factor 1 alpha 2 and pS2. The remaining 18 clones did not match sequences in the GenBank/EMBL database, indicating that they may be novel genes. Expression of pS2, neuropeptide Y receptor Y1 and three novel clones was induced by estradiol, indicating estrogen-responsiveness. The expression pattern of one novel gene, DEME-6, correlated with expression of ER and ERF-1/AP-2 gamma in a panel of breast carcinoma cell lines. A 2.6 kb cDNA of DEME-6 was sequenced and contains an open reading frame of 574 amino acids that demonstrates 62.4% similarity with a gene from Caenorhabditis elegans chromosome III. Expression of DEME-6 was also detected in primary breast carcinomas but not in normal breast tissue, as determined by RT-PCR. These findings support the hypothesis that a set of genes coordinately regulated with ER, but not necessarily estradiol-responsive, are characteristic of the hormone-responsive breast cancer phenotype.