miR-100-3p inhibits cell proliferation and induces apoptosis in human gastric cancer through targeting to BMPR2
miR-100-3p inhibits cell proliferation and induces apoptosis in human gastric cancer through targeting to BMPR2
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DOI：
10.1186/s12935-019-1060-2
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发表时间：
2019-12-27
影响因子：
5.8
通讯作者：
Wang, Si-Ying
中科院分区：
文献类型：
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作者：
Peng, Chun-Wei;Yue, Ling-Xiao;Wang, Si-Ying
研究背景miR-100与胃癌的发生、发展密切相关。然而，miR-100- 3 p在GC中的潜在机制仍不清楚。在本研究中，我们打算研究miR-100- 3 p如何调节GC恶性肿瘤。方法采用实时荧光定量PCR（qRT-PCR）检测miR-100- 3 p在体外（胃癌细胞系GES-1和胃癌细胞系GC）和体内（胃癌组织和正常胃组织）的表达水平。MTT和PE/Annexin V分析负责测量miR-100- 3 p对GC细胞增殖和凋亡的影响。Transwell法检测有无基质胶作用下胃癌细胞的迁移和侵袭能力。通过转录组学分析和荧光素酶报告基因分析证实miR-100- 3 p与骨形态发生蛋白受体2（BMPR 2）的相互作用。采用qRT-PCR和Western blot方法检测ERK/AKT和Bax/Bcl 2/Caspase 3在miR-100- 3 p功能障碍中的表达。结果miR-100- 3 p在胃癌细胞系和癌组织中表达下调，与BMPR 2表达呈负相关。miR-100- 3 p的缺失促进肿瘤生长和BMPR 2表达。一致地，miR-100- 3 p抑制对GC细胞的作用被BMPR 2的敲低部分中和。过表达miR-100- 3 p可同时抑制肿瘤生长和下调BMPR 2表达。因此，BMPR 2的过表达部分地中和了miR-100- 3 p过表达的作用。进一步的研究表明BMPR 2介导miR-100- 3 p的下游效应，可能间接调控ERK/AKT和Bax/Bcl 2/Caspase 3信号通路。结论miR-100- 3 p作为一种抑癌miRNA，通过下调BMPR 2的表达，抑制ERK/AKT信号通路，激活Bax/Bcl 2/Caspase 3信号通路。这一发现为GC提供了新的见解，并可能有助于确定新的诊断和治疗靶点。
Background miR-100 has been reported to closely associate with gastric cancer (GC) initiation and progression. However, the underlying mechanism of miR-100-3p in GC is still largely unclear. In this study, we intend to study how miR-100-3p regulates GC malignancy. Methods The expression levels of miR-100-3p in vitro (GES-1 and GC cell lines) and in vivo (cancerous and normal gastric tissues) were examined by quantitative real-time PCR (qRT-PCR). MTT and PE/Annexin V analyses were responsible for measurement of the effects of miR-100-3p on GC cell proliferation and apoptosis. Transwell assay with or without matrigel was used to examine the capacity of migration and invasion in GC cells. The interaction of miR-100-3p with bone morphogenetic protein receptor 2 (BMPR2) was confirmed through transcriptomics analysis and luciferase reporter assay. qRT-PCR and Western blot analyses were applied to determine the expression of ERK/AKT and Bax/Bcl2/Caspase3, which were responsible for the dysfunction of miR-100-3p. Results miR-100-3p was down-regulated in GC cell lines and cancerous tissues, and was negatively correlated with BMPR2. Loss of miR-100-3p promoted tumor growth and BMPR2 expression. Consistently, the effects of miR-100-3p inhibition on GC cells were partially neutralized by knockdown of BMPR2. Over-expression of miR-100-3p simultaneously inhibited tumor growth and down-regulated BMPR2 expression. Consistently, over-expression of BMPR2 partially neutralized the effects of miR-100-3p over-expression. Further study demonstrated that BMPR2 mediated the effects downstream of miR-100-3p, which might indirectly regulate ERK/AKT and Bax/Bcl2/Caspase3 signaling pathways. Conclusion miR-100-3p acted as a tumor-suppressor miRNA that down-regulated BMPR2, which consequently inhibited the ERK/AKT signaling and activated Bax/Bcl2/Caspase3 signaling. This finding provided novel insights into GC and could contribute to identify a new diagnostic and therapeutic target.