穿心莲内酯联合平阳霉素对人口腔鳞癌细胞SCC-9凋亡的影响

Effects of andrographolide and pingyangmycin on apoptosis of human oral squamous cell carcinoma cell line SCC-9

目的探讨穿心莲内酯是否有增强平阳霉素抗人口腔鳞癌细胞的作用,并探讨其潜在作用机制。方法 MTT法检测不同浓度的穿心莲内酯与平阳霉素联合应用对人口腔鳞癌细胞SCC-9细胞活力的影响,确定最佳的浓度组合;流式细胞术、划痕实验分别检测细胞凋亡率及迁徙能力;蛋白质免疫印迹(Western blot)法分析相应的凋亡机制。结果 50μmol·L-1的穿心莲内酯和平阳霉素联合发挥最佳作用,流式细胞术及划痕实验结果显示穿心莲内酯联合平阳霉素的促凋亡作用、抗迁徙能力强于两者单独应用; Western blot结果显示与单独应用穿心莲内酯和平阳霉素相比,两者联合显著提高了凋亡相关蛋白BAX、天冬氨酸半胱氨酸蛋白酶-3(caspase-3)、caspase-9的表达。结论穿心莲内酯对平阳霉素的促SCC-9细胞凋亡作用具有协同效应,并通过caspase-9通路发挥促凋亡作用。

Objective To investigate the synergistic antitumor activity of andrographolide(ANDRO)in combination with pingyangmycin(PYM)in OSCC and the enhanced antitumor activity mechanisms.Methods The combinatorial effect of ANDRO and PYM on the proliferation of OSCC cell line SCC-9 was evaluated using MTT assays.Scratch wound test was carried out to examine cell migration and flow cytometry was conducted to analyze cell apoptosis.Western blot analysis was used to investigate Bcl-2,Bax and caspase proteins,as well as the expression of Stat3 proteins in SCC-9 cells.Results 50μmol·L-1 ANDRO combined with PYM exerted the best anti-proliferation effect.Flow cytometry and scratches experimental results showed that the pro-apoptotic effect,anti-migration ability of ANDRO/PYM combination were stronger than the two groups alone.Western blot showed that ANDRO/PYM combination significantly increased the expression of Bax,caspase-3 and caspase-9 compared with either agent alone.Conclusions Our results indicated ANDRO enhances the antitumor effect of PYM through caspase-9-dependent mitochondrial pathways.