Gopalan Vinod, Smith Robert A, Lam Alfred K-Y
Cancer Molecular Pathology, School of Medicine and Griffith Health Institute, Griffith University, Gold Coast, QLD, Australia.
Cancer Molecular Pathology, School of Medicine and Griffith Health Institute, Griffith University, Gold Coast, QLD, Australia.
Exp Cell Res. 2015 Jan 15;330(2):423-428. doi: 10.1016/j.yexcr.2014.08.006. Epub 2014 Aug 13.
miR-498 is a non-coding RNA located intergenically in 19q13.41. Due to its predicted targeting of several genes involved in control of cellular growth, we examined the expression of miR-498 in colon cancer cell lines and a large cohort of patients with colorectal adenocarcinoma. Two colon cancer cancer cell lines (SW480 and SW48) and one normal colonic epithelial cell line (FHC) were recruited. The expression of miR-498 was tested in these cell lines by using quantitative real-time polymerase chain reaction (qRT-PCR). Tissues from 80 patients with surgical resection of colorectum (60 adenocarcinomas and 20 non-neoplastic tissues) were tested for miR-498 expression by qRT-PCR. In addition, an exogenous miR-498 (mimic) was used to detect the miRNA׳s effects on cell proliferation and cell cycle events in SW480 using MTT calorimetric assay and flow cytometry respectively. The colon cancer cell lines showed reduced expression of miR-498 compared to a normal colonic epithelial cell line. Mimic driven over expression of miR-498 in the SW480 cell line resulted in reduced cell proliferation and increased proportions of G2-M phase cells. In tissues, miR-498 expression was too low to be detected in all colorectal adenocarcinoma compared to non-neoplastic tissues. This suggests that the down regulation of miR-498 in colorectal cancer tissues and the direct suppressive cellular effect noted in cancer cell lines implies that miR-498 has some direct or indirect role in the pathogenesis of colorectal adenocarcinomas.
miR-498是一种位于19q13.41基因间区域的非编码RNA。由于其预测可靶向调控多个参与细胞生长控制的基因,我们检测了miR-498在结肠癌细胞系以及一大群大肠腺癌患者中的表达情况。我们选取了两种结肠癌细胞系(SW480和SW48)以及一种正常结肠上皮细胞系(FHC)。通过定量实时聚合酶链反应(qRT-PCR)检测这些细胞系中miR-498的表达。采用qRT-PCR检测了80例接受结肠直肠手术切除患者的组织(60例腺癌组织和20例非肿瘤组织)中miR-498的表达。此外,分别使用MTT比色法和流式细胞术,通过外源性miR-498(模拟物)检测其对SW480细胞增殖和细胞周期事件的影响。与正常结肠上皮细胞系相比,结肠癌细胞系中miR-498的表达降低。在SW480细胞系中,模拟物驱动的miR-498过表达导致细胞增殖减少,G2-M期细胞比例增加。在组织中,与非肿瘤组织相比,所有大肠腺癌组织中miR-498的表达均过低而无法检测到。这表明大肠癌细胞组织中miR-498的下调以及在癌细胞系中观察到的直接抑制细胞效应意味着miR-498在大肠腺癌的发病机制中具有一些直接或间接作用。