白花蛇舌草对人舌鳞癌细胞Tca83的作用机制研究

Mechanical study of Hedyotis diffusa on tongue squamous cell carcinoma celline Tca83 in vitro

目的通过体外实验研究不同浓度白花蛇舌草提取液作用不同时间后对人口腔舌鳞癌细胞株Tca83凋亡的影响。方法配制0、100、300、600和900μg/ml 5种浓度的白花蛇舌草提取物,分别作用24、48和72 h后,通过免疫组化法、MTT法及流式细胞仪检测Tca83细胞的存活率、Bcl-2蛋白表达及凋亡情况。结果各浓度组的白花蛇舌草提取物均可促进Tca83细胞不同程度凋亡,并观察到抗凋亡的Bcl-2蛋白减少或消失;随着白花蛇舌草提取液浓度的增加和作用时间的延长,细胞凋亡率随之增加;低浓度白花蛇舌草提取液在短时间作用后,将Tca83细胞抑制在G0/G1期;所有浓度组白花蛇舌草提取液经过长时间作用于Tca83细胞后,将其抑制在G2/M期。结论白花蛇舌草在体外实验中可显著促进Tca83细胞凋亡,并且在一定作用时间及药物浓度范围内,细胞凋亡程度与药物作用时间和药物浓度成正比。

Objective To explore the effect of different concentrations of Hedyotis diffusa on tongue squamous cell carcinoma cell line Tca83 about apoptosis in vitro.MethodsTca83 cells were treated by Hedyotis diffusa with different concentrations(0,100,300,600 and 900μg/ml)and different time(24 h,48 h,72 h)in vitro.The induction of apoptosis of Tca83 cells was observed with immunohistochemical staining,MTT assay and flow cytometry.Results Various concentrations of Hedyotis diffusa could promote Tca83 cells apoptosis,and the expression of anti-apoptotic protein Bcl-2 was downregulated or disappeared.The apoptosis rate increased with the increase of concentration and the prolongation of treatment time.Obviously,Tca83 cells showed GO/Gl arrest in low concentration of Hedyotis diffusa,and they showed G2/M arrest after drug action for a long time.Conclusions Hedyotis diffusa can significantly promote the apoptosis of Tca83 cells.Within a certain range of treating time and dose,the degree of apoptosis is proportional to the time and concentration of the drug.