Department of Thoracic Surgery, Tongren Hospital, Shanghai Jiao Tong University, School of Medicine, Shanghai 200336, P.R. China.
Department of Thoracic Surgery, Tongren Hospital, Shanghai Jiao Tong University, School of Medicine, Shanghai 200336, P.R. China
Biosci Rep. 2019 Feb 1;39(2). doi: 10.1042/BSR20181523. Print 2019 Feb 28.
The present study aims to investigate the mechanism of miR-384 in non-small cell lung cancer (NSCLC) cell apoptosis and autophagy by regulating Collagen α-1(X) chain (COL10A1). Bioinformatics methods were applied to evaluate potential miRNAs and genes that might correlate with NSCLC. Tumor tissues and adjacent tissues from 104 NSCLC patients were collected and human NSCLC A549 cell line was selected for subsequent experiments. A549 cells were treated with miR-384 mimic, miR-384 inhibitor, or knockdown of COL10A1. Quantitative real-time PCR (qRT-PCR) and Western blotting were utilized to detect the levels of miR-384, COL10A, Survivin, Bcl-2, Bax, Bcl-xl, Beclin 1, and LC3 in tissues and cells. A series of biological assays including MTT assay, Annexin V-FITC/PI (propidium iodide) staining, immunofluorescence, monodansylcadaverine (MDC) staining were conducted to investigate the effects of miR-384 and COL10A1 on NSCLC cells. Tumorigenicity assay for nude rats was applied. Results obtained from the present study indicated that miR-384 down-regulated COL10A1 by targetting it. Compared with adjacent tissues, miR-384 expression was obviously reduced while COL10A1 expression was significantly enhanced in NSCLC tissues (all <0.05). Outcomes and suggested that cell proliferation and tumorigenicity were inhibited while cell apoptosis and autophagy were induced in NSCLC cells treated with up-regulation of miR-384 or silence of COL10A1. In miR-384 inhibitor group, cell proliferation was improved, while cell apoptosis was reduced and cell autophagy was decreased whereas tumorigenicity of cells was strengthened. Based on the findings of our study, it was established that miR-384 could down-regulate COL10A1 levels, subsequently inhibiting cell proliferation and promoting cell apoptosis and autophagy in NSCLC cells.
本研究旨在通过调控 Collagen α-1(X) 链（COL10A1）来探讨 miR-384 在非小细胞肺癌（NSCLC）细胞凋亡和自噬中的作用机制。采用生物信息学方法评估可能与 NSCLC 相关的潜在 miRNA 和基因。收集了 104 例 NSCLC 患者的肿瘤组织和相邻组织，并选择人 NSCLC A549 细胞系进行后续实验。用 miR-384 模拟物、miR-384 抑制剂或 COL10A1 敲低处理 A549 细胞。运用实时定量 PCR（qRT-PCR）和 Western blot 检测组织和细胞中 miR-384、COL10A、Survivin、Bcl-2、Bax、Bcl-xl、Beclin 1 和 LC3 的水平。通过 MTT 测定、Annexin V-FITC/PI（碘化丙啶）染色、免疫荧光、单丹磺酰尸胺（MDC）染色等一系列生物学实验研究 miR-384 和 COL10A1 对 NSCLC 细胞的影响。并进行裸鼠成瘤实验。研究结果表明，miR-384 通过靶向 COL10A1 下调其表达。与相邻组织相比，miR-384 在 NSCLC 组织中表达明显降低，而 COL10A1 表达明显升高（均<0.05）。结果表明，上调 miR-384 或沉默 COL10A1 可抑制 NSCLC 细胞增殖，促进细胞凋亡和自噬。在 miR-384 抑制剂组中，细胞增殖改善，细胞凋亡减少，细胞自噬减少，而细胞致瘤性增强。根据本研究的结果，miR-384 可下调 COL10A1 水平，进而抑制 NSCLC 细胞增殖，促进细胞凋亡和自噬。
