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Comparison of Circulating miRNAs Expression Alterations in Matched Tissue and Plasma Samples During Colorectal Cancer Progression

机译:结直肠癌进展中匹配组织和血浆样品中循环miRNA表达改变的比较

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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 (p0.05). 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.
机译:已经发现MicroRNAS(miRNA)在结肠直肠腺瘤 - 癌序列中发挥着关键作用。 MiRNA特异性高通量阵列变得可用于检测有希望的miRNA表达改变,即使在生物流体,例如血浆样品,其中miRNA是稳定的。该研究的目的是鉴定循环miRNA,显示正常结肠(N),管状腺瘤(ADT),小管腺瘤(ADTV)和结直肠癌(CRC)匹配的血浆和组织样品之间的改变的表达。六个外周等离子体和匹配的组织活检样品(n n = 4; ADT n = 4; ADTV n = 4; CRC N = 4),分离包括miRNA分数的总RNA。使用QIAAMP循环核酸试剂盒(QIAGEN)提取来自等离子体样品的miRNA。匹配的组织血浆MiRNA微阵列实验通过GeneChip miRNA 3.0阵列(Affymetrix)进行。 RT-QPCR(MicroRNA即用型PCR人类面板I + II; EXIQON)用于验证。在血浆样品中的AD和CRC基团(miR-149 *,miR-3196,miR-4687)比较中观察到特征miRNA表达改变。在N与CRC比较中,通过RT-PCR检测miR-612,miR-1296,miR-933,miR-937和miR-1207的显着过表达(P <0.05)。使用微阵列在组织对中观察这些miRNA的类似表达模式。尽管与组织相比,在较低浓度的循环系统中也发现miRNA,但表达模式略微重叠在组织和等离子体样品之间。检测到循环miRNA改变不仅可以来自引物肿瘤,而是来自包括免疫细胞的其他细胞类型。

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