Gastrointestinal Cancer Research Laboratory, Baylor University Medical Center, Dallas, TX 75246, USA.
Cancer Epidemiol Biomarkers Prev. 2010 Jul;19(7):1766-74. doi: 10.1158/1055-9965.EPI-10-0027. Epub 2010 Jun 15.
Colorectal cancer (CRC) is the second leading cause of cancer-related deaths, but currently available noninvasive screening programs have achieved only a modest decrease in mortality. MicroRNAs (miRNA) play an important role in a wide array of biological processes and are commonly dysregulated in neoplasia. We aimed to evaluate the feasibility of fecal miRNAs as biomarkers for colorectal neoplasia screening.
Total RNA was extracted from freshly collected stool samples from 8 healthy volunteers and 29 samples collected via fecal occult blood testing from subjects with normal colonoscopies, colon adenomas, and CRCs. miRNA expression analyses were done with TaqMan quantitative reverse transcription-PCR for a subset of miRNAs. Illumina miRNA microarray profiling was done to evaluate the differences in expression patterns between normal colonic mucosa tissues and stool samples from healthy subjects.
We efficiently extracted miRNAs from stool specimens using our developed protocol. Data from independent experiments showed high reproducibility for miRNA extraction and expression. miRNA expression patterns were similar in stool specimens among healthy volunteers, and reproducible in stool samples that were collected serially in time from the same individuals. miRNA expression profiles from 29 patients showed higher expression of miR-21 and miR-106a in patients with adenomas and CRCs compared with individuals free of colorectal neoplasia.
Our data indicate that miRNAs could be extracted from stool easily and reproducibly. The stools of patients with colorectal neoplasms have unique and identifiable patterns of miRNA expression.
Fecal miRNAs may be an excellent candidate for the development of a noninvasive screening test for colorectal neoplasms.
结直肠癌(CRC)是癌症相关死亡的第二大主要原因,但目前可用的非侵入性筛查计划仅实现了死亡率的适度降低。microRNAs(miRNA)在广泛的生物过程中发挥着重要作用,并且在肿瘤中通常失调。我们旨在评估粪便 miRNA 作为结直肠肿瘤筛查生物标志物的可行性。
从 8 名健康志愿者和 29 名通过粪便潜血试验采集的样本中提取新鲜采集的粪便样本中的总 RNA,这些样本来自接受正常结肠镜检查、结肠腺瘤和 CRC 的受试者。使用 TaqMan 定量逆转录 PCR 对一组 miRNA 进行 miRNA 表达分析。进行 Illumina miRNA 微阵列分析,以评估正常结肠黏膜组织与健康受试者粪便样本之间的表达模式差异。
我们使用我们开发的方案从粪便标本中有效提取 miRNA。独立实验的数据表明 miRNA 提取和表达的重现性很高。健康志愿者的粪便标本中 miRNA 表达模式相似,并且从同一个体中随时间连续采集的粪便标本中具有可重复性。29 名患者的 miRNA 表达谱显示,腺瘤和 CRC 患者的 miR-21 和 miR-106a 表达较高,而无结直肠肿瘤的个体则较低。
我们的数据表明,miRNA 可以从粪便中轻松且可重复地提取出来。结直肠肿瘤患者的粪便具有独特且可识别的 miRNA 表达模式。
粪便 miRNA 可能是开发结直肠肿瘤非侵入性筛查试验的绝佳候选物。