Cancer Institute (Key Laboratory of Cancer Prevention and Intervention, China National Ministry of Education, Key Laboratory of Molecular Biology in Medical Sciences, Zhejiang Province, China), The Second Affiliated Hospital, Zhejiang University School of Medicine, 88 Jiefang Road, Hangzhou 310009, Zhejiang, China.
Med Oncol. 2012 Jun;29(2):1037-43. doi: 10.1007/s12032-011-9975-z. Epub 2011 May 12.
Human microRNA-9 (miR-9) has been reported to be involved in the metastasis of several malignancies including brain breast cancer. However, its role in the metastasis of colorectal cancer (CRC) remains to be revealed. Here, we evaluated miR-9 expression in metastatic CRC and investigated its effects on the motility and proliferation of RKO cells. The expressions of miR-9 in 15 primary CRC specimens without distant metastasis (NM group) and 10 primary CRC specimens (M group) with distant metastasis (M group) were determined by quantitative real-time PCR. The alternations in the motility and morphology of RKO cells before and after miR-9 transfection were analyzed by migration assay and F-actin staining. The relationship between miR-9 and α-catenin was identified by Western blotting. Cell growth was examined by MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazoliumbromide) assay. Significant difference of miR-9 expression was observed in M group compared to the NM group (P < 0.001). Ectopic expression of miR-9 enhanced the motility of RKO cells as well as changed their morphological appearance, while cell growth remained unchanged. The overexpression of miR-9 could also down-regulate α-catenin expression. These data suggest that miR-9 may potentially participate in the metastatic process of CRC though facilitating cell motility.
人 microRNA-9 (miR-9) 已被报道参与包括脑乳腺癌在内的几种恶性肿瘤的转移。然而,其在结直肠癌 (CRC) 转移中的作用仍有待揭示。在这里,我们评估了转移性 CRC 中 miR-9 的表达,并研究了其对 RKO 细胞迁移和增殖的影响。通过定量实时 PCR 测定了 15 例无远处转移 (NM 组) 和 10 例有远处转移 (M 组) 的原发性 CRC 标本中 miR-9 的表达。通过迁移实验和 F-肌动蛋白染色分析 miR-9 转染前后 RKO 细胞迁移和形态的变化。通过 Western blot 鉴定 miR-9 与α-连环蛋白的关系。通过 MTT (3-[4,5-二甲基噻唑-2-基]-2,5-二苯基四氮唑溴盐) 实验检测细胞生长。M 组 miR-9 的表达明显低于 NM 组 (P < 0.001)。miR-9 的异位表达增强了 RKO 细胞的迁移能力,并改变了它们的形态外观,而细胞生长保持不变。miR-9 的过表达还可以下调α-连环蛋白的表达。这些数据表明,miR-9 可能通过促进细胞迁移而潜在地参与 CRC 的转移过程。