Department of Respiratory Medicine, The Second Affiliated Hospital of Soochow University, Suzhou, Jiangsu Province, China.
Department of Respiratory Medicine, The Second Affiliated Hospital of Soochow University, Suzhou, Jiangsu Province, China.
Kaohsiung J Med Sci. 2018 Nov;34(11):593-605. doi: 10.1016/j.kjms.2018.06.005. Epub 2018 Jul 5.
The aim of this study was to investigate the role of microRNA-26b (miR-26b) in regulating the proliferation, migration, and apoptosis of small cell lung cancer (SCLC) cells. First, we examined the expression level of miR-26b in human normal fetal lung fibroblasts (NFLFs) and three SCLC cell lines NCI-H466, NCI-H1688, and NCI-H196. In the following experiments, the three SCLC cell lines were transfected with miR-26b mimic and inhibitor. Cell growth and survival, as well as migration and invasion capacities were determined by MTT, colony formation, Transwell migration and invasion, and wound healing assays. Cell apoptosis, production of reactive oxygen species, and mitochondrial membrane potential were also measured in the three cell lines following various treatments. As a result, we found that the level of miR-26b was significantly lower in SCLC cells than in NFLFs. Additionally, transfection with miR-26b mimic could inhibit proliferation, colony formation, and migration, as well as induce apoptosis in these SCLC cell lines; while miR-26b inhibitor showed the opposite effects. Further mechanistic experiment revealed that miR-26b could suppress the expression of myeloid cell leukemia 1 protein (Mcl-1) and the 3'-untranslated region (3'-UTR) of Mcl-1 may be the direct binding site of miR-26b, suggesting that the effect of miR-26b may be mediated by targeting Mcl-1. Collectively, our findings offer a new insight into the role of miR-26b in the pathogenesis of SCLC, and provide primary evidence supporting the potential of miR-26b-based therapy for the treatment of SCLC.
本研究旨在探讨 microRNA-26b(miR-26b)在调节小细胞肺癌(SCLC)细胞增殖、迁移和凋亡中的作用。首先,我们检测了 miR-26b 在人正常胎儿肺成纤维细胞(NFLFs)和三种 SCLC 细胞系 NCI-H466、NCI-H1688 和 NCI-H196 中的表达水平。在后续实验中,我们将这三种 SCLC 细胞系分别转染 miR-26b 模拟物和抑制剂。通过 MTT、集落形成、Transwell 迁移和侵袭以及划痕愈合实验检测细胞生长和存活以及迁移和侵袭能力。还在三种细胞系中测量了细胞凋亡、活性氧(ROS)产生和线粒体膜电位。结果发现,SCLC 细胞中 miR-26b 的水平明显低于 NFLFs。此外,转染 miR-26b 模拟物可抑制这些 SCLC 细胞系的增殖、集落形成和迁移,并诱导细胞凋亡;而 miR-26b 抑制剂则表现出相反的效果。进一步的机制实验表明,miR-26b 可抑制髓样细胞白血病 1 蛋白(Mcl-1)的表达,且 Mcl-1 的 3'-非翻译区(3'-UTR)可能是 miR-26b 的直接结合位点,表明 miR-26b 的作用可能是通过靶向 Mcl-1 介导的。综上所述,本研究结果为 miR-26b 在 SCLC 发病机制中的作用提供了新的见解,并为基于 miR-26b 的治疗 SCLC 的潜在应用提供了初步证据。