雷公藤甲素对舌鳞癌细胞自噬的影响及相关机制研究

The effects of triptolide on autophagy of human tongue squamous cell carcinoma cells and the related mechanism of PIM1-STAT3 signaling pathway

目的:研究雷公藤甲素(TP)对人舌鳞状细胞癌细胞(TSCCs)自噬的影响及相关机制。方法:用2.5~20μg/L的TP处理体外培养的TSCCs CAL-27和SCC-25,MTT实验检测细胞活力,MDC染色检测细胞中自噬泡,流式细胞术检测细胞凋亡,Western blot法检测细胞中自噬相关蛋白,凋亡相关蛋白,STAT3-PIM-1信号通路蛋白表达情况,并通过加入自噬抑制剂检测自噬与凋亡的关系。结果:TP能够诱导TSCC细胞发生自噬,并且对肿瘤细胞有明显的凋亡诱导作用,促进凋亡蛋白PARP和Caspase-9剪切体的表达,同时下调SATA3-PIM-1蛋白表达,加入自噬抑制剂后细胞凋亡发生率减少。结论:TP可能通过抑制STAT3-PIM-1信号通路诱导TSCCs发生自噬,并进一步诱导细胞凋亡,发挥抗肿瘤作用。

Objective:To investigate the effects of triptolide(TP)on autophagy of human tongue squamous cell carcinoma cells(TSCCs)and its related mechanism.Methods:TSCC cells of CAL-27 and SCC-25 were cultured in vitro and treated with 2.5-20μg/L TP,the cell viability was detected by MTT assay,autophagic vacuoles in the cells were detected by MDC staining,and cell apoptosis was detected by flow cytometry.Western blot method was used to detect the expression of autophagy-related proteins,apoptosis-related proteins and STAT3-PIM-1 signaling pathway proteins in the cells.The relationship between autophagy and apoptosis was detected by adding autophagy inhibitors.Results:TP induced autophagy of the cancer cells and showed apoptosis-inducing effect on the cells.TP promoted the expression of apoptotic proteins PARP and Caspase-9 spliceosome,and down-regulated SATA3-PIM-1 protein expression,the incidence of apoptosis was reduced after adding the autophagy inhibitor.Conclusion:TP may induce autophagy of tongue cancer cells by inhibiting the STAT3-PIM-1 signaling pathway,and further induce cell apoptosis to play an anti-tumor effect.