Department of Pharmacy, College of Pharmacy, CHA University, Yatap-dong, Bundang-gu, Seongnam-si, Gyeonggi-do 463836, Korea.
Mol Cancer Res. 2013 Jan;11(1):43-53. doi: 10.1158/1541-7786.MCR-12-0355. Epub 2012 Oct 18.
MicroRNAs (miRNA) are small noncoding RNAs that regulate gene expression in human diseases, including lung cancer. miRNAs have oncogenic and nononcogenic functions in lung cancer. In this study, we report the identification of a novel miRNA, miR-7515, from lung cancer cells. The novel miR-7515 was characterized using various predictive programs and experimental methods. miR-7515 was able to forming a stem-loop structure and its sequence was conserved in mammals. The expression level of miR-7515 in lung cancer cells and tissues was profiled using TaqMan miRNA assays. miR-7515 was downregulated in lung cancer compared with normal human lung cells and tissues. The target of miR-7515 was determined using a dual luciferase reporter assay. Expression of the target gene was determined by quantitative RT-PCR and Western blot analysis after transfection with miR-7515. miR-7515 directly suppressed human mesenchymal-epithelial transition factor (c-Met) by binding to the 3' untranslated region (UTR). Overexpression of miR-7515 significantly decreased cell-cycle-related proteins downstream of c-Met through c-Met inhibition. Cell proliferation and migration were examined using the XTT proliferation assay and the Transwell migration assay. miR-7515 led to decreased cell proliferation, migration and invasion in a lung cancer cell line. These results suggest that miR-7515 plays an important role in the proliferation and migration of lung cancer cells through c-Met regulation.
微小 RNA(miRNA)是一种小的非编码 RNA,可调节人类疾病中的基因表达,包括肺癌。miRNA 在肺癌中有致癌和非致癌功能。在这项研究中,我们从肺癌细胞中报告了一种新型 miRNA,miR-7515 的鉴定。新型 miR-7515 采用各种预测程序和实验方法进行了特征描述。miR-7515 能够形成茎环结构,其序列在哺乳动物中保守。采用 TaqMan miRNA 检测法对肺癌细胞和组织中的 miR-7515 表达水平进行了分析。miR-7515 在肺癌中与正常人类肺细胞和组织相比下调。通过双荧光素酶报告基因检测确定了 miR-7515 的靶基因。转染 miR-7515 后通过定量 RT-PCR 和 Western blot 分析测定靶基因的表达。miR-7515 通过结合 3'非翻译区(UTR)直接抑制人间质上皮转化因子(c-Met)。通过抑制 c-Met,过表达 miR-7515 显著下调 c-Met 下游的细胞周期相关蛋白。通过 XTT 增殖测定和 Transwell 迁移测定检查细胞增殖和迁移。miR-7515 导致肺癌细胞系中的细胞增殖、迁移和侵袭减少。这些结果表明,miR-7515 通过 c-Met 调节在肺癌细胞的增殖和迁移中发挥重要作用。