School of Life Science and Technology, China Pharmaceutical University, Nanjing, People's Republic of China.
PLoS One. 2012;7(5):e35435. doi: 10.1371/journal.pone.0035435. Epub 2012 May 31.
MicroRNAs (miRNAs) are a class of small noncoding RNAs that regulate gene expression by targeting mRNAs to trigger either translation repression or mRNA degradation. miR-125b is down-regulated in human breast cancer cells compared with the normal ones except highly metastatic tumor cells MDA-MB-231. However, few functional studies were designed to investigate metastatic potential of miR-125b. In this study, the effects of miR-125b on metastasis in human breast cancer cells were studied, and the targets of miR-125b were also explored. Transwell migration assay, cell wound healing assay, adhesion assay and nude mice model of metastasis were utilized to investigate the effects of miR-125b on metastasis potential in vitro and in vivo. In addition, it was implied STARD13 (DLC2) was a direct target of miR-125b by Target-Scan analysis, luciferase reporter assay and western blot. Furthermore, activation of STARD13 was identified responsible for metastasis induced by miR-125b through a siRNA targeting STARD13. qRT-PCR, immunofluorescent assay and western blot was used to observe the variation of Vimentin and α-SMA in breast cancer cells. In summary, our study provided new insights into the function of miR-125b during the metastasis of breat cancer cells and also suggested the role of miR-125b in pro-metastasis by targeting STARD13.
微小 RNA(miRNAs)是一类小的非编码 RNA,通过靶向 mRNAs 来触发翻译抑制或 mRNA 降解,从而调节基因表达。与正常细胞相比,人乳腺癌细胞中的 miR-125b 下调,除了高度转移性肿瘤细胞 MDA-MB-231 之外。然而,很少有功能研究设计来研究 miR-125b 的转移潜能。在这项研究中,研究了 miR-125b 对人乳腺癌细胞转移的影响,并探索了 miR-125b 的靶标。利用 Transwell 迁移测定、细胞划痕愈合测定、黏附测定和裸鼠转移模型,研究了 miR-125b 对体外和体内转移潜能的影响。此外,通过靶标扫描分析、荧光素酶报告测定和 Western blot 暗示 STARD13(DLC2)是 miR-125b 的直接靶标。此外,通过靶向 STARD13 的 siRNA 鉴定,激活 STARD13 是 miR-125b 诱导转移所必需的。qRT-PCR、免疫荧光测定和 Western blot 用于观察乳腺癌细胞中波形蛋白和α-SMA 的变化。总之,我们的研究提供了对乳腺癌细胞转移过程中 miR-125b 功能的新见解,并表明 miR-125b 通过靶向 STARD13 在促进转移中发挥作用。