Guo F, Lin S C, Zhao M S, Yu B, Li X Y, Gao Q, Lin D J
Department of Respiratory Medicine, Shandong Provincial Hospital Affiliated to Shandong University, Jinan, Shandong, China.
Department of Respiratory Medicine, Yantai Affiliated Hospital of Binzhou Medical University, Yantai, Shandong, China.
Braz J Med Biol Res. 2017 Jul 3;50(7):e5974. doi: 10.1590/1414-431X20175974.
microRNA (miR)-142-3p is implicated in malignancy and has been identified as a biomarker for aggressive and recurrent lung adenocarcinomas. This study aimed to evaluate the inhibitory effect of miR-142-3p on apoptosis and inflammation induced by bleomycin in MLE-12 cells. MLE-12 cells were first transfected either with miR-142-3p mimic or miR-142-3p inhibitor and then the cells were exposed to 50 μg/mL of bleomycin. Thereafter, cell viability, apoptosis and the expression of pro-inflammatory cytokines were assessed using CCK-8, flow cytometry, RT-PCR and western blot analyses. Cox-2, PI3K, AKT and mTOR expressions were detected by western blotting after bleomycin was administered together with NS-398 (an inhibitor of Cox-2). As a result, cell viability was significantly decreased, as well as apoptosis and the expression of IL-1 and TNF-α were remarkably increased after 50 and 100 μg/mL of bleomycin administration. miR-142-3p overexpression alleviated bleomycin-induced apoptosis and overproduction of these two pro-inflammatory cytokines, while miR-142-3p suppression exhibited completely opposite results. Up-regulation of Cox-2 and inactivation of PI3K/AKT/mTOR were found in bleomycin-pretreated cells, while these abnormal regulations were partially abolished by miR-142-3p overexpression and NS-398. In conclusion, this study demonstrated that miR-142-3p overexpression protected bleomycin-induced injury in lung epithelial MLE-12 cells, possibly via regulating Cox-2 expression and PI3K/AKT/mTOR signaling pathway. These findings provide evidence that miR-142-3p may be a therapeutic strategy for idiopathic pulmonary fibrosis (IPF) treatment.
微小RNA(miR)-142-3p与恶性肿瘤有关,已被确定为侵袭性和复发性肺腺癌的生物标志物。本研究旨在评估miR-142-3p对博来霉素诱导的MLE-12细胞凋亡和炎症的抑制作用。首先用miR-142-3p模拟物或miR-142-3p抑制剂转染MLE-12细胞,然后将细胞暴露于50μg/mL的博来霉素中。此后,使用CCK-8、流式细胞术、RT-PCR和蛋白质印迹分析评估细胞活力、凋亡和促炎细胞因子的表达。在给予博来霉素的同时给予NS-398(一种Cox-2抑制剂)后,通过蛋白质印迹检测Cox-2、PI3K、AKT和mTOR的表达。结果,在给予50和100μg/mL博来霉素后,细胞活力显著降低,凋亡以及IL-1和TNF-α的表达显著增加。miR-142-3p过表达减轻了博来霉素诱导的凋亡和这两种促炎细胞因子的过量产生,而miR-142-3p抑制则表现出完全相反的结果。在博来霉素预处理的细胞中发现Cox-2上调和PI3K/AKT/mTOR失活,而这些异常调节被miR-142-3p过表达和NS-398部分消除。总之,本研究表明miR-142-3p过表达可保护博来霉素诱导的肺上皮MLE-12细胞损伤,可能是通过调节Cox-2表达和PI3K/AKT/mTOR信号通路。这些发现提供了证据表明miR-142-可能是特发性肺纤维化(IPF)治疗的一种策略。