Identification of differentially expressed genes in human prostate cancer using subtraction and microarray.
Identification of differentially expressed genes in human prostate cancer using subtraction and microarray.
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发表时间：
2000-03
期刊：
影响因子：
11.2
通讯作者：
Jiangchun Xu;J. Stolk;Xinqun Zhang;Sandra J. Silva;Raymond L. Houghton;Masazumi Matsumura;T. Vedvick;Kevin B. Leslie;Roberto Badaró;Steven G. Reed
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作者：
Jiangchun Xu;J. Stolk;Xinqun Zhang;Sandra J. Silva;Raymond L. Houghton;Masazumi Matsumura;T. Vedvick;Kevin B. Leslie;Roberto Badaró;Steven G. Reed
我们已经使用cDNA文库减法结合高通量微阵列筛选鉴定了人类前列腺癌和组织特异性基因。生成前列腺肿瘤和正常前列腺组织的cDNA文库。减文库的特征显示了癌症和组织特异性基因的富集。通过集落杂交消除了高度冗余的克隆。剩余的克隆被选择用于微阵列，以确定各种肿瘤和正常组织中的基因表达水平。选择在前列腺肿瘤和/或正常前列腺组织中显示过表达的克隆并进行测序。在这里，我们报告了两个基因的鉴定，P503S和P504S，从减法文库和第三个基因，P510S，通过减法和微阵列筛选。Northern blot、real-time PCR （TaqMan）和免疫组化进一步证实其表达谱在前列腺组织和/或前列腺肿瘤中过表达。克隆全长cDNA序列，利用生物信息学算法PSORT预测其亚细胞位置为质膜蛋白。通过这些方法鉴定的基因是癌症诊断和治疗的潜在候选者。
We have identified human prostate cancer- and tissue-specific genes using cDNA library subtraction in conjunction with high throughput microarray screening. Subtracted cDNA libraries of prostate tumors and normal prostate tissue were generated. Characterization of subtracted libraries showed enrichment of both cancer- and tissue-specific genes. Highly redundant clones were eliminated by colony hybridization. The remaining clones were selected for microarray to determine gene expression levels in a variety of tumor and normal tissues. Clones showing overexpression in prostate tumors and/or normal prostate tissues were selected and sequenced. Here we report the identification of two genes, P503S and P504S, from subtracted libraries and a third gene, P510S, by subtraction followed by microarray screening. Their expression profiles were further confirmed by Northern blot, real-time PCR (TaqMan), and immunohistochemistry to be overexpressed in prostate tissues and/or prostate tumors. Full-length cDNA sequences were cloned, and their subcellular locations were predicted by a bioinformatic algorithm, PSORT, to be plasma membrane proteins. The genes identified through these approaches are potential candidates for cancer diagnosis and therapy.