Clinical applications of microarray technology: creatine kinase B is an up-regulated gene in epithelial ovarian cancer and shows promise as a serum marker
Clinical applications of microarray technology: creatine kinase B is an up-regulated gene in epithelial ovarian cancer and shows promise as a serum marker
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DOI：
10.1016/j.ygyno.2004.08.047
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发表时间：
2005-01-01
影响因子：
4.7
通讯作者：
Mok, SC
中科院分区：
文献类型：
--
作者：
Huddleston, HG;Wong, KK;Mok, SC
Objective. (1)为了鉴定和（2）验证在卵巢癌中上调的基因，和（3）调查与良性盆腔肿块患者和正常对照相比，卵巢癌患者术前血清中肌酸激酶B（CK B）的活性是否升高。应用MICROMAX基因芯片系统和卵巢癌细胞株及正常卵巢上皮细胞（HOSE）的RNA，筛选差异表达基因。使用来自两种细胞系和通过激光捕获显微切割（LCM）获得的组织的RNA，我们进行定量PCR以验证这些基因之一肌酸激酶B（CK B）的上调。使用市售的酶测定法，CKB活性测定术前血清样品中获得的45例卵巢癌患者，49例良性盆腔肿块，以及37名正常对照。采用受损学生t检验进行统计学分析。微阵列技术显示，CKB基因表达的癌症HOSE比率为18。通过实时PCR测量的CKB的RNA水平，在癌细胞系中与HOSE细胞相比升高了36倍（8.4），在显微切割的卵巢癌上皮细胞中与正常卵巢上皮细胞相比升高了22.75倍（10.45）。在血清中，癌症患者CKB酶活性的平均标准误为24.7 U/L（+/-5.1），良性肿块患者为9.6 U/L（+/-1.6）（P = 0.0088），正常对照组为8.5 U/L（1.7）（P = 0.0096）。微阵列技术提供了一种鉴定具有潜在临床用途的肿瘤生物标志物的方法。我们的数据表明，CKB基因表达上调卵巢癌细胞在体外和体内，CKB酶活性显着升高，从卵巢癌患者的血清中，包括那些与1期疾病。这些发现表明CKB作为早期诊断标志物的潜在作用。(C)2004年爱思唯尔公司All rights reserved.
Objective. (1) To identify and (2) validate genes that are up-regulated in ovarian cancer, and (3) to investigate whether the activity of a candidate gene, creatine kinase B (CKB) is elevated in pre-operative sera from ovarian cancer patients compared to patients with benign pelvic masses and normal controls.Methods. MICROMAX cDNA microarray system and RNA derived from pooled ovarian cancer cell lines and normal ovary surface epithelial cells (HOSE) were used to identify differentially expressed genes. Using RNA from both cell lines and from tissue obtained through laser capture microdissection (LCM), we performed quantitative PCR in order to validate up-regulation of one of these genes, creatine kinase B (CKB). Using a commercially available enzyme assay, CKB activity was measured in pre-operative serum samples obtained from 45 ovarian cancer patients, 49 patients with a benign pelvic mass, as well as 37 normal controls. Statistical analysis was preformed using an impaired Student's t test.Results. Microarray technology revealed that CKB gene expression had a cancer to HOSE ratio of 18. RNA levels of CKB, measured by real-time PCR, were elevated a mean (and standard error) of 36-fold (8.4) in cancer cell lines compared with HOSE cells and 22.75-fold (10.45) in microdissected ovarian cancer epithelial cells compared with normal ovarian epithelial cells. In serum, the mean standard error) of CKB enzyme activity in cancer cases was 24.7 U/L units (+/-5.1) compared to 9.6 U/L (+/-1.6) for benign mass cases (P = 0.0088) and to 8.5 U/L (1.7) for normal controls (P = 0.0096).Conclusions. Microarray technology offers a method to identify tumor biomarkers with potential clinical usefulness. Our data indicated that CKB gene expression is up-regulated in ovarian cancer cells in vitro and in vivo and that CKB enzyme activity is significantly elevated in sera from ovarian cancer patients, including those with stage 1 disease. These findings suggest a potential role for CKB as a marker for early diagnosis. (C) 2004 Elsevier Inc. All rights reserved.