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微小 RNA-466(miR-466)在结直肠癌(CRC)中作为肿瘤抑制因子和预后因素发挥作用。

MicroRNA-466 (miR-466) functions as a tumor suppressor and prognostic factor in colorectal cancer (CRC).

机构信息

Department of Surgery, Lanxi People's Hospital, Zhejiang, China.

出版信息

Bosn J Basic Med Sci. 2018 Aug 1;18(3):252-259. doi: 10.17305/bjbms.2018.2376.

Abstract

MicroRNAs (miRNAs) have an important role in the regulation of tumor development and metastasis. In this study, we investigated the clinical and prognostic value as well as biological function of miR-466 in colorectal cancer (CRC). Tumor and adjacent healthy tissues were obtained from 100 patients diagnosed with CRC. miR-466 expression was determined by quantitative reverse transcription polymerase chain reaction (qRT-PCR). mRNA and protein levels of cyclin D1, apoptosis regulator BAX (BAX), and matrix metalloproteinase-2 (MMP-2) were analyzed by qRT-PCR and Western blot, respectively, in SW-620 CRC cells transfected with miR-466 mimics or negative control miRNA. Effects of miR-466 on SW-620 cell proliferation, cell cycle and apoptosis, and invasion were investigated using CCK-8 assay, flow cytometry and Transwell assay, respectively. miR-466 expression was significantly downregulated in tumor tissues compared to matched adjacent non-tumor tissues. Low expression of miR-466 was significantly correlated with the tumor size, Tumor Node Metastasis stage, lymph node metastasis, and distant metastasis. The overall survival of CRC patients with low miR-466 expression was significantly shorter compared to high-miR-466 expression group (log-rank test: p = 0.0103). Multivariate analysis revealed that low miR-466 expression was associated with poor prognosis in CRC patients. The ectopic expression of miR-466 suppressed cell proliferation and migration/invasion, as well as induced G0/G1 arrest and apoptosis in SW-620 cells. Moreover, the ectopic expression of miR-466 decreased the expression of cyclin D1 and MMP-2, but increased BAX expression in SW-620 cells. In conclusion, our findings demonstrated that miR-466 functions as a suppressor miRNA in CRC and may be used as a prognostic factor in these patients.

摘要

微小 RNA(miRNAs)在肿瘤发生和转移的调控中具有重要作用。在这项研究中,我们研究了 miR-466 在结直肠癌(CRC)中的临床和预后价值以及生物学功能。从 100 名被诊断为 CRC 的患者中获得肿瘤和相邻健康组织。通过定量逆转录聚合酶链反应(qRT-PCR)测定 miR-466 的表达。在转染 miR-466 模拟物或阴性对照 miRNA 的 SW-620 CRC 细胞中,通过 qRT-PCR 和 Western blot 分别分析细胞周期蛋白 D1、凋亡调节因子 BAX(BAX)和基质金属蛋白酶-2(MMP-2)的 mRNA 和蛋白水平。使用 CCK-8 测定法、流式细胞术和 Transwell 测定法分别研究 miR-466 对 SW-620 细胞增殖、细胞周期和凋亡以及侵袭的影响。与匹配的相邻非肿瘤组织相比,肿瘤组织中 miR-466 的表达明显下调。miR-466 表达的低水平与肿瘤大小、肿瘤淋巴结转移分期、淋巴结转移和远处转移显著相关。miR-466 表达低的 CRC 患者的总生存期明显短于 miR-466 表达高的患者(对数秩检验:p=0.0103)。多变量分析显示,miR-466 低表达与 CRC 患者预后不良相关。miR-466 的异位表达抑制了 SW-620 细胞的增殖和迁移/侵袭,并诱导了 G0/G1 期阻滞和凋亡。此外,miR-466 的异位表达降低了 SW-620 细胞中细胞周期蛋白 D1 和 MMP-2 的表达,但增加了 BAX 的表达。总之,我们的研究结果表明,miR-466 在 CRC 中作为一种抑制性 miRNA 发挥作用,可作为这些患者的预后因素。

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