BACKGROUND Recent studies have shown 6'-O-galloylpaeoniflorin (GPF), a nature product extracted from the roots of paeoniflorin exerts anti-oxidant and anti-inflammatory activities. However, the effects of GPF on the proliferation and invasion in non-small cell lung cancer (NSCLC) cells have not been clarified. METHODS MTT assay was performed to determine the cytotoxicity of GPF treatment on NSCLC cells. Colony formation assay, cell scratch test and transwell assay were performed to determine the proliferation and invasion of NSCLC cells in vitro, respectively. An A549 cell xenograft mouse model was performed to confirm the growth of NSCLC cells in vivo. Western blotting was used to measure the levels of activating transcription factor 2 (ATF2), AMP-activated protein kinase (AMPK) and phosph-AMPK (p-AMPK). Luciferase assay was used to validate the binding of miR-299-5p on the 3' untranslated region (UTR) of ATF2. RESULTS Administration of GPF (50 or 100 μM) was significantly cytotoxic to A549 cells and H1299 cells, as well as inhibited the clonality, invasion and metastasis of NSCLC cells in vitro. GPF treatment also inhibited the tumor growth of NSCLC cell mouse xenografts in vivo. Exotic expression of miR-299-5p significantly inhibited the growth of NSCLC cells in vitro and in vivo. Downregulation of miR-299-5p expression attenuated the inhibition of the proliferation and metastasis of non-small cell lung cancer cells by GPF treatment. miR-299-5p significantly decreased ATF2 mRNA and protein levels in A549 cells (p < 0.05). Overexpression of ATF2 blocked the inhibitory effect of miR-299-5p on the proliferation and invasiveness of A549 cells. CONCLUSIONS GPF regulates miR-299-5p/ATF2 axis in A549 cells via the AMPK signalling pathway, thereby inhibiting the proliferation and metastasis of non-small cell lung cancer cells.

中文翻译：

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6'-O-galloylpaeoniflorin通过AMPK / miR-299-5p / ATF2轴调节非小细胞肺癌的增殖和转移。

背景技术最近的研究表明，从pa药苷的根中提取的天然产物6'-O-半乳糖苷酸（GPF）具有抗氧化和消炎的作用。但是，GPF对非小细胞肺癌（NSCLC）细胞增殖和侵袭的影响尚不清楚。方法采用MTT法测定GPF处理对NSCLC细胞的细胞毒性。进行集落形成测定，细胞刮擦试验和transwell测定，分别测定NSCLC细胞的体外增殖和侵袭。进行A549细胞异种移植小鼠模型以证实NSCLC细胞在体内的生长。Western印迹法用于测量活化转录因子2（ATF2），AMP活化蛋白激酶（AMPK）和磷酸化AMPK（p-AMPK）的水平。使用萤光素酶测定法验证miR-299-5p在ATF2的3'非翻译区（UTR）上的结合。结果GPF（50或100μM）的给药对A549细胞和H1299细胞具有明显的细胞毒性，并在体外抑制了NSCLC细胞的克隆性，侵袭和转移。GPF治疗还抑制了NSCLC细胞小鼠异种移植物的体内肿瘤生长。miR-299-5p的外源表达在体外和体内均显着抑制了NSCLC细胞的生长。通过GPF处理，miR-299-5p表达的下调减弱了对非小细胞肺癌细胞增殖和转移的抑制作用。miR-299-5p显着降低A549细胞中ATF2 mRNA和蛋白水平（p <0.05）。ATF2的过表达阻止了miR-299-5p对A549细胞增殖和侵袭性的抑制作用。结论GPF通过AMPK信号通路调节A549细胞中的miR-299-5p / ATF2轴，从而抑制非小细胞肺癌细胞的增殖和转移。