MicroRNA-148b regulates tumor growth of non-small cell lung cancer through targeting MAPK/JNK pathway
MicroRNA-148b regulates tumor growth of non-small cell lung cancer through targeting MAPK/JNK pathway
复制标题
MicroRNA-148b通过靶向MAPK/JNK通路调控非小细胞肺癌肿瘤生长
DOI：
10.1186/s12885-019-5400-3
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发表时间：
2019-03-08
期刊：
影响因子：
3.8
通讯作者：
Liu, Guolong
中科院分区：
文献类型：
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作者：
Lu, Lin;Liu, Qiyao;Liu, Guolong
研究背景MicroRNA-148 b（miR-148 b）在多种肿瘤中被检测到，被普遍认为是肿瘤抑制因子。我们前期的研究发现miR-148 b在人非小细胞肺癌（NSCLC）标本和细胞系中表达降低。然而，miR-148 b调控肿瘤生长的机制尚不清楚。方法首先通过动物实验验证miR-148 b是否具有抑制肿瘤生长的作用。然后，通过将含有miR-148 B模拟物或阴性对照（NC）模拟物（shRNA对照）的重组质粒转染到NSCLC细胞系PC 14/B和A549细胞中来研究潜在的机制。用未包装的慢病毒载体转染的肿瘤细胞用作空白对照。CCK-8试剂盒和集落形成实验检测细胞增殖能力。比较细胞周期阻滞以阐明肿瘤细胞增殖的机制。Annexin V-FITC凋亡检测试剂盒检测miR-148 b对细胞凋亡的影响。结果miR 148 b过表达可明显抑制肿瘤生长，而敲低miR 148 b则可明显促进肿瘤生长。进一步的实验表明miR-148 b抑制肿瘤细胞增殖。此外，miR 148 b的过表达可阻止NSCLC细胞进入有丝分裂期，从而减少G2/M期细胞数，促进肿瘤细胞凋亡。结论miR-148 b通过阻断MAPK/JNK信号通路，抑制NSCLC细胞增殖，诱导凋亡，从而抑制肿瘤生长。
BackgroundMicroRNA-148b (miR-148b) has been detected in various types of tumors, and is generally viewed as a tumor suppressor. Our previous study found the decreased expression of miR-148b in human non small cell lung cancer (NSCLC) specimens and cell lines. However, the underlying mechanisms of miR-148b in regulating tumor progression remain unclear.MethodsFirstly animal experiments were performed to verify whether miR-148b could inhibit the tumor growth. Then, the underlying mechanisms were studied by transfecting recombinant plasmids containing a miR-148b mimic or a negative control (NC) mimic (shRNA control) into NSCLC cell lines PC14/B and A549 cells. Tumor cells transfected with unpackaged lentiviral vectors was used as blank control. Cell proliferation capabilities were measured by using CCK-8 kit and colony formation assay. Cell cycle arrest was compared to clarify the mechanism underlying the tumor cell proliferation. Annexin V-FITC Apoptosis Detection kit was applied to investigate the effect of miR-148b on cell apoptosis. Furthermore, western blot analysis were performed to study the targeting pathway.ResultsWe found that over-expression of miR148b could significantly inhibit tumor growth, while knocking down miR148b could obviously promote tumor growth. Further experiment showed that miR-148b inhibited tumor cell proliferation. Besides, over-expression of miR148b decreased the G2/M phase population of the cell cycle by preventing NSCLC cells from entering the mitotic phase and enhanced tumor cell apoptosis. Further western blot analysis indicated that miR148b could inhibit mitogen-activated protein kinase/Jun N-terminal kinase (MAPK/JNK) signaling by decreasing the expression of phosphorylated (p) JNK.ConclusionsThese results demonstrate that miR-148b could inhibit the tumor growth and act as tumor suppressor by inhibiting the proliferation and inducing apoptosis of NSCLC cells by blocking the MAPK/JNK pathway.