surfactin对舌鳞癌Tca8113细胞株增殖、凋亡和迁移作用的影响

Effect of Surfactin on Proliferation,Apoptosis and Migration of Tongue Squamous Cell Carcinoma Tca8113 Cell Line

目的初步探讨枯草芽孢杆菌发酵产物surfactin对舌鳞癌Tca8113细胞的增殖、凋亡及迁移作用的影响。方法体外培养舌鳞癌Tca8113细胞,加入不同浓度梯度的药物,分别培养24 h、48 h后,SRB(磺酰罗丹明B)法检测surfactin对舌鳞癌细胞的增殖抑制作用;平板克隆形成实验,检测surfactin对舌鳞癌Tca8113细胞生长能力的影响;Hochest/PI双染法观察surfactin对舌鳞癌Tca8113细胞形态变化的影响;Annexin-V-FITC/PI双染流式细胞仪检测surfactin对舌鳞癌Tca8113细胞调亡的影响;wound healing实验观察药物作用前后肿瘤细胞迁移距离的变化;免疫印迹法验证基质金属蛋白酶MMPs表达的情况。结果 SRB结果分析,surfactin能够有效抑制舌鳞癌细胞的增殖,并随药物浓度的升高和作用时间的延长抑制率不断升高,呈时间-剂量依赖性,24 h、48 h的半数抑制浓度(IC50)分别为74 ug/ml和53 ug/ml;平板克隆实验结果显示surfactin能够抑制舌鳞癌Tca8113细胞的克隆形成,药物组与对照组比较具有统计学意义(P<0.05);Hochest/PI双染法、Annexin-V-FITC/PI双染流式细胞仪检测得出surfactin能够诱导舌鳞癌Tca8113细胞凋亡;wound healing实验观察得到,surfactin可明显阻碍舌鳞癌细胞划痕后的愈合能力;免疫印迹法结果得出surfactin可以通过下调MMPs蛋白表达抑制舌鳞癌Tca8113细胞迁移。结论 surfactin能够明显抑制舌鳞癌Tca8113细胞的增殖、迁移并有促凋亡的作用。

Objective Preliminarily investigate the effect of Bacillus subtilis fermentation products surfactin on proliferation, apoptosis and migration of tongue squamous cell carcinoma TcaS113 cell line. Methods The tongue squamous cell carcinoma TcaS113 cell were cuhured in vitro,then treated with different concentrations of surfactin. The proliferation effect on tongue squamous cell carcinoma Tea8113 cells were checked by Sulforhodamine B （SRB） assay when the cells were cultured 24h and 48h; Clonogenic assay was used to detected the effect of surfactin on the growth of tongue squamous cell carcinoma Tca8113 cell ;Morphological changes of tongue squamous cell carcinoma Tea8113 cell was observed by Hoehest/PI staining fluorescence microscope;Cell apoptosis effect was measured by flow eytometry;The change of the migration distance of tongue squamous cell carcinoma TeaS113 cell was observed by wound healing assay;The expression of matrix metalloprotei- nase MMPs was verified by Western blot assay. Results The SRB assay displayed that surfactin can effectively inhabit the proliferation of tongue squamous cell carcinoma Tca8113 cell with dose-and time-dependent manner, and the IC50 were 74ug/ml and 53ug/ml for 24 and 48 hours respectively; Clonogenic assay results showed that surfactin could inhibit the tongue squamous cell carcinoma Tca8113 cell clone formation, The drug group had statistical significance （P 〈 0.05 ） compared with the control group;Hochest/PI staining,Annexin-V-FITC/PI double staining flow cytometry revealed that Surfactin could induce apoptosis of tongue squamous cell carcinoma Tca8113 cell . Healing wound assay observed that surfactin can significantly inhibit the healing of cell scratches;Western blotting results showed that surfaetin can inhibit TeaS113 cell migration by down-regulating the expression of MMPs . Conclusion Surfactin can significantly inhibit the proliferation, migration and pro-apoptosis of tongue squamous cell carcinoma cell Tca8113.