通过汇集样本表达谱分析，骨桥蛋白被确定为结肠癌进展的主要标志物。

Osteopontin identified as lead marker of colon cancer progression, using pooled sample expression profiling.

作者信息

Agrawal Deepak, Chen Tingan, Irby Rosalyn, Quackenbush John, Chambers Ann F, Szabo Marianna, Cantor Alan, Coppola Domenico, Yeatman Timothy J

机构信息

Department of Cell Biology, Interdisciplinary Oncology, H. Lee Moffitt Cancer Center, University of South Florida, Tampa 33612, USA.

出版信息

J Natl Cancer Inst. 2002 Apr 3;94(7):513-21. doi: 10.1093/jnci/94.7.513.

DOI:10.1093/jnci/94.7.513

PMID:11929952

Abstract

BACKGROUND

New tumor markers and markers of tumor progression are needed for improved staging and for better assessment of treatment of many cancers. Gene expression profiling techniques offer the opportunity to discover such markers. We investigated the feasibility of sample pooling strategy in combination with a novel analysis algorithm to identify markers.

METHODS

Total RNA from human colon tumors (n = 60) of multiple stages (adenomas; cancers with modified Astler Collier stages B, C, and D; and liver metastases) were pooled within stages and compared with pooled normal mucosal specimens (n = 10) by using oligonucleotide expression arrays. Genes that showed consistent increases or decreases in their expression through tumor progression were identified. Northern blot analysis was used to validate the findings. All statistical tests were two-sided.

RESULTS

More than 300 candidate tumor markers and more than 100 markers of tumor progression were identified. Northern analysis of 11 candidate tumor markers confirmed the gene expression changes. The gene for the secreted integrin-binding protein osteopontin was most consistently differentially expressed in conjunction with tumor progression. Its potential as a progression marker was validated (Spearman's rho = 0.903; P<.001) with northern blot analysis using RNA from an independent set of 10 normal and 43 tumor samples representing all stages. Moreover, a statistically significant correlation between osteopontin protein expression and advancing tumor stage was identified with the use of 303 additional specimens (human cancer = 185, adenomas = 67, and normal mucosal specimens = 51) (Spearman's rho = 0.667; P<.001).

CONCLUSIONS

Sample pooling can be a powerful, cost-effective, and rapid means of identifying the most common changes in a gene expression profile. We identified osteopontin as a clinically useful marker of tumor progression by use of gene expression profiling on pooled samples.

摘要

背景

为了改善多种癌症的分期以及更好地评估治疗效果，需要新的肿瘤标志物和肿瘤进展标志物。基因表达谱分析技术为发现此类标志物提供了机会。我们研究了样本合并策略与一种新型分析算法相结合以识别标志物的可行性。

方法

将来自多个阶段（腺瘤；改良阿斯特勒 - 科利尔分期为B、C和D期的癌症；以及肝转移瘤）的60例人类结肠肿瘤的总RNA在各阶段内进行合并，并通过寡核苷酸表达阵列与合并的正常黏膜标本（n = 10）进行比较。识别出在肿瘤进展过程中表达持续增加或减少的基因。采用Northern印迹分析来验证研究结果。所有统计检验均为双侧检验。

结果

识别出300多个候选肿瘤标志物和100多个肿瘤进展标志物。对11个候选肿瘤标志物的Northern分析证实了基因表达变化。分泌型整合素结合蛋白骨桥蛋白的基因在肿瘤进展过程中差异表达最为一致。通过对来自另一组独立的10例正常样本和43例代表所有阶段的肿瘤样本进行Northern印迹分析，验证了其作为进展标志物的潜力（Spearman秩相关系数 = 0.903；P <.001）。此外，使用另外303份标本（人类癌症 = 185例，腺瘤 = 67例，正常黏膜标本 = 51例）确定骨桥蛋白蛋白表达与肿瘤进展期之间存在统计学显著相关性（Spearman秩相关系数 = 0.667；P <.001）。