Inhibition of lung cancer growth and metastasis by DHA and its metabolite, RvD1, through miR-138-5p/FOXC1 pathway
Inhibition of lung cancer growth and metastasis by DHA and its metabolite, RvD1, through miR-138-5p/FOXC1 pathway
复制标题
DHA 及其代谢物 RvD1 通过 miR-138-5p/FOXC1 通路抑制肺癌生长和转移
DOI：
10.1186/s13046-019-1478-3
复制
发表时间：
2019-11-29
影响因子：
11.3
通讯作者：
Pan, Jinshun
中科院分区：
文献类型：
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作者：
Bai, Xiaoming;Shao, Jiaofang;Pan, Jinshun
非小细胞肺癌（non-small cell lung cancer，NSCLC）是世界上最常见的恶性肿瘤之一。已知DHA能够抑制NSCLC细胞增殖和转移。然而，DHA显示其抗肿瘤作用的机制尚不清楚。本研究旨在探讨DHA及其代谢产物对肺癌细胞生长和侵袭的影响及其机制。方法采用体外细胞活力测定和transwell法检测细胞增殖和侵袭能力。将omega-6 PUFA转化为omega-3 PUFA的转基因mfat-1小鼠用于检测内源性DHA对肿瘤移植的影响。LC-MS/MS分析鉴定了DHA的几种类二十烷酸代谢物的升高。通过qPCR miRNA芯片、在线预测软件、荧光素酶报告基因分析和Western blot分析，进一步阐明了其作用机制。结果外源性DHA可抑制NSCLC细胞的生长和侵袭。内源性产生的DHA减弱转基因mfat-1小鼠中LLC衍生的肿瘤生长和转移。在DHA代谢产物的升高中，Resolvin D1（RvD 1）显著抑制细胞生长和侵袭。miRNA芯片显示，RvD 1处理后，miR-138- 5 p的表达水平显著升高。miR-138- 5 p模拟物降低细胞活力和侵袭;而miR-138- 5 p抑制剂消除RvD 1介导的细胞活力和侵袭抑制。过表达miR-138- 5 p后，FOXC 1的表达显著降低，而荧光素酶报告基因检测显示FOXC 1是miR-138- 5 p的直接靶点。在体内，通过mfat-1转基因的内源性DHA增强了miR-138- 5 p的表达并降低了FOXC 1的表达。此外，FOXC 1的过表达逆转了RvD 1处理诱导的细胞活力和侵袭力的抑制。结论RvD 1/miR-138- 5 p/FOXC 1通路是DHA及其代谢产物RvD 1作用于NSCLC的一种新机制，靶向该通路可能成为治疗NSCLC的一种新策略。
Background Non small cell lung cancer (NSCLC) is one of the most common cancers in the world. DHA is known to be capable of suppressing NSCLC cell proliferation and metastasis. However, the mechanisms by which DHA exhibits its antitumor effects are unknown. Here we aimed to identify the effects and mechanisms of DHA and its metabolites on lung cancer cell growth and invasion. Methods As measures of cell proliferation and invasion ability, the cell viability and transwell assays were used in vitro. Transgenic mfat-1 mice, which convert omega-6 PUFAs to omega-3 PUFAs, were used to detect the effect of endogenous DHA on tumor transplantation. An LC - MS/MS analysis identified the elevation of several eicosanoid metabolites of DHA. By using qPCR miRNA microarray, online prediction software, luciferase reporter assays and Western blot analysis, we further elucidated the mechanisms. Results Addition of exogenous DHA inhibited the growth and invasion in NSCLC cells in vitro. Endogenously produced DHA attenuated LLC-derived tumor growth and metastasis in the transgenic mfat-1 mice. Among the elevation of DHA metabolites, resolvin D1 (RvD1) significantly contributed to the inhibition in cell growth and invasion. MiRNA microarray revealed that the level of miR-138-5p was significantly increased after RvD1 treatment. MiR-138-5p mimics decreased cell viability and invasion; while miR-138-5p inhibitor abolished RvD1-mediated suppression of cell viability and invasion. The expression of FOXC1 was significantly reduced upon overexpression of miR-138-5p while luciferase reporter assay showed that FOXC1 was a direct target of miR-138-5p. In vivo, endogenous DHA by the mfat-1 transgene enhanced miR-138-5p expression and decreased FOXC1 expression. Furthermore, overexpression of FOXC1 reversed the inhibition in cell viability and invasion induced by RvD1 treatment. Conclusions These data identified the RvD1/miR-138-5p/FOXC1 pathway as a novel mechanism by DHA and its metabolite, RvD1, and the potential of targeting such pathway as a therapeutic strategy in treating NSCLC.