Nagy Zsófia Brigitta, Barták Barbara Kinga, Kalmár Alexandra, Galamb Orsolya, Wichmann Barnabás, Dank Magdolna, Igaz Péter, Tulassay Zsolt, Molnár Béla
Molecular Gastroenterology Laboratory, 2nd Department of Internal Medicine, Semmelweis University, Szentkirályi street 46, Budapest, 1088, Hungary.
Molecular Medicine Research Group, Hungarian Academy of Sciences, Budapest, Hungary.
Pathol Oncol Res. 2019 Jan;25(1):97-105. doi: 10.1007/s12253-017-0308-1. Epub 2017 Oct 4.
MicroRNAs (miRNAs) have been found to play a critical role in colorectal adenoma-carcinoma sequence. MiRNA-specific high-throughput arrays became available to detect promising miRNA expression alterations even in biological fluids, such as plasma samples, where miRNAs are stable. The purpose of this study was to identify circulating miRNAs showing altered expression between normal colonic (N), tubular adenoma (ADT), tubulovillous adenoma (ADTV) and colorectal cancer (CRC) matched plasma and tissue samples. Sixteen peripheral plasma and matched tissue biopsy samples (N n = 4; ADT n = 4; ADTV n = 4; CRC n = 4) were selected, and total RNA including miRNA fraction was isolated. MiRNAs from plasma samples were extracted using QIAamp Circulating Nucleic Acid Kit (Qiagen). Matched tissue-plasma miRNA microarray experiments were conducted by GeneChip® miRNA 3.0 Array (Affymetrix). RT-qPCR (microRNA Ready-to-use PCR Human Panel I + II; Exiqon) was used for validation. Characteristic miRNA expression alterations were observed in comparison of AD and CRC groups (miR-149*, miR-3196, miR-4687) in plasma samples. In the N vs. CRC comparison, significant overexpression of miR-612, miR-1296, miR-933, miR-937 and miR-1207 was detected by RT-PCR (p < 0.05). Similar expression pattern of these miRNAs were observed using microarray in tissue pairs, as well. Although miRNAs were also found in circulatory system in a lower concentration compared to tissues, expression patterns slightly overlapped between tissue and plasma samples. Detected circulating miRNA alterations may originate not only from the primer tumor but from other cell types including immune cells.
微小RNA（miRNA）已被发现，在结直肠腺瘤-癌序列中起着关键作用。即使在生物体液（如血浆样本，其中miRNA稳定）中，也可使用特定于miRNA的高通量阵列来检测有前景的miRNA表达改变。本研究的目的是鉴定在正常结肠（N）、管状腺瘤（ADT）、绒毛状管状腺瘤（ADTV）和结直肠癌（CRC）匹配的血浆和组织样本之间显示表达改变的循环miRNA。选择了16份外周血浆和匹配的组织活检样本（N n = 4；ADT n = 4；ADTV n = 4；CRC n = 4），并分离了包括miRNA部分的总RNA。使用QIAamp循环核酸试剂盒（Qiagen）从血浆样本中提取miRNA。通过GeneChip® miRNA 3.0阵列（Affymetrix）进行匹配的组织-血浆miRNA微阵列实验。使用RT-qPCR（miRNA即用型PCR人试剂盒I + II；Exiqon）进行验证。在血浆样本中比较AD组和CRC组（miR-149*、miR-3196、miR-4687）时，观察到了特征性的miRNA表达改变。在N与CRC的比较中，通过RT-PCR检测到miR-612、miR-1296、miR-933、miR-937和miR-1207显著过表达（p < 0.05）。在组织对中使用微阵列也观察到了这些miRNA的相似表达模式。尽管与组织相比，在循环系统中也发现miRNA的浓度较低，但组织和血浆样本之间的表达模式略有重叠。检测到的循环miRNA改变可能不仅源于原发肿瘤，还源于包括免疫细胞在内的其他细胞类型。
