microRNA-148b 通过靶向 MAPK/JNK 通路调节非小细胞肺癌的肿瘤生长。
MicroRNA-148b regulates tumor growth of non-small cell lung cancer through targeting MAPK/JNK pathway.
机构信息
Department of Medical Oncology, Guangzhou First People's Hospital, Guangzhou Medical University, Guangzhou, 510180, Guangdong, China.
Department of Medical Oncology, Guangzhou First People's Hospital, School of Medicine, South China University of Technology, Guangzhou, 510180, Guangdong, China.
出版信息
BMC Cancer. 2019 Mar 8;19(1):209. doi: 10.1186/s12885-019-5400-3.
BACKGROUND
MicroRNA-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.
METHODS
Firstly 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.
RESULTS
We 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.
CONCLUSIONS
These 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.
背景
miR-148b（微小 RNA-148b）已在多种类型的肿瘤中被检测到，通常被视为肿瘤抑制因子。我们之前的研究发现，miR-148b 在人非小细胞肺癌（NSCLC）标本和细胞系中的表达降低。然而，miR-148b 调节肿瘤进展的潜在机制尚不清楚。
方法
首先进行动物实验以验证 miR-148b 是否能抑制肿瘤生长。然后，通过转染含有 miR-148b 模拟物或阴性对照（shRNA 对照）的重组质粒，研究 NSCLC 细胞系 PC14/B 和 A549 细胞中的潜在机制。未包装的慢病毒载体转染的肿瘤细胞作为空白对照。使用 CCK-8 试剂盒和集落形成实验测量细胞增殖能力。比较细胞周期停滞以阐明肿瘤细胞增殖的机制。应用 Annexin V-FITC Apoptosis Detection kit 研究 miR-148b 对细胞凋亡的影响。此外，进行 Western blot 分析以研究靶向途径。
结果
我们发现过表达 miR148b 可显著抑制肿瘤生长，而敲低 miR148b 则可明显促进肿瘤生长。进一步的实验表明，miR-148b 抑制肿瘤细胞增殖。此外，过表达 miR148b 通过阻止 NSCLC 细胞进入有丝分裂期，降低细胞周期的 G2/M 期细胞群，增强肿瘤细胞凋亡。进一步的 Western blot 分析表明，miR148b 通过降低磷酸化（p）JNK 的表达抑制丝裂原激活蛋白激酶/Jun N 末端激酶（MAPK/JNK）信号通路。
结论
这些结果表明，miR-148b 通过抑制 MAPK/JNK 通路，抑制 NSCLC 细胞的增殖并诱导其凋亡，从而抑制肿瘤生长并发挥肿瘤抑制作用。
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