南蛇藤提取物通过DJ-1/PTEN/FAK轴抑制非小细胞肺癌侵袭转移

Nansheteng(Celastrus orbiculatus Thunb.)Extract Inhibits Non-Small Cell Lung Cancer Invasion and Metastasis viaDJ-1/PTEN/FAK Axis

目的研究南蛇藤提取物(Celastrus orbiculatus Thunb.Extract,COE)抑制非小细胞肺癌侵袭转移的具体机制。方法通过CCK-8法测定COE抑制H1299细胞的生物活性,并依此选取低毒浓度进行细胞侵袭转移的功能研究。使用伤口愈合实验和高内涵成像追踪细胞轨迹对细胞运动功能进行测定。通过Western bolt实验进行分子层面的研究,探究COE抑制肺癌侵袭转移的具体分子机制。实验的分组采用剂量梯度进行设置,分别为Control组(0.05%DMSO),COE组:20,40,80μg/mL。结果COE以剂量依赖的形式明显抑制了H1299的细胞活性。伤口愈合实验结果显示,COE显著抑制了H1299细胞的迁徙能力,统计分析显示差异有显著统计学意义。高内涵成像实时动态追踪细胞轨迹显示,80μg/mL浓度的COE显著抑制了H1299细胞的迁徙能力,表现在运动速度的降低和均方位移量的下降。Western bolt实验结果表明,COE通过DJ-1/PTEN/FAK轴抑制非小细胞侵袭转移。蛋白质印迹的归一化分析均具有统计学意义。结论南蛇藤提取物能在低细胞毒浓度下通过DJ-1/PTEN/FAK轴抑制非小细胞侵袭转移。

Objective To investigate the specific mechanism of inhibition of invasion and metastasis of non-small cell lung cancer by Nancheteng(Celastrus orbiculatus Thunb.)extract(COE).Methods The biological activity of COE inhibition in H1299 cells was determined by CCK-8 assay and the low toxic concentration was selected accordingly for the functional study of cell invasion and metastasis.The cell motility function was measured using wound healing assay and high-content imaging to track cell trajectories.Molecular studies were performed by Western bolt assay to investigate the specific molecular mechanism of COE inhibition of lung cancer invasion and metastasis.The grouping of the experiments was set up using a dose gradient for the control group(0.05% DMSO)and COE group(20μg/mL,40μg/mL and 80μg/mL).Results COE significantly inhibited the cellular activity of H1299 in a dose-dependent manner.The results of wound healing assay showed that COE significantly inhibited the migratory ability of H1299 cells,and the statistical analysis showed a significant difference.Real-time dynamic tracking of cell trajectories by high-content imaging showed that COE significantly inhibited the migratory ability of H1299 cells by the concentration of 80μg/mL,as evidenced by reduced migration speed and decreased mean square displacement.Western bolt assays showed that COE inhibited non-small cell invasive metastasis via the DJ-1/PTEN/FAK axis.The normalized analysis of protein blots were all statistically significant.Conclusion COE inhibits non-small cell lung cancer invasion and metastasis via DJ-1/PTEN/FAK axis at low cytotoxic concentrations.