乳杆菌A-2代谢产物诱导人舌鳞癌CAL-27细胞凋亡的实验研究

发布时间：2018-10-08 08:06

【摘要】：目的：探讨乳杆菌A-2代谢产物对人舌鳞癌CAL-27细胞的增殖抑制与诱导凋亡的作用。方法：乳杆菌A-2分别经复原乳清培养基和复原乳清菊糖培养基培养，制备相应的代谢产物1（LSPM1）和代谢产物2(LSPM2)；采用MTT法检测对CAL-27细胞的体外增殖抑制作用；通过吖啶橙染色，荧光显微镜下观察细胞形态学改变；采用流式细胞仪和DNA琼脂凝胶电泳法检测细胞凋亡情况。结果：在LSPM1和LSPM2在3.0mg/mL~48mg/mL的区间内，随着代谢产物浓度的增加，对CAL-27细胞的抑制作用逐渐增强。LSPM1和LSPM2的半抑制浓度(IC50)分别为15.1mg/mL和10.27mg/mL；荧光显微镜下观察，凋亡细胞的细胞核不规则，细胞皱缩，，凝聚呈新月形或马蹄形，贴壁不良；流式细胞仪结果显示：经LSPM1和LSPM2处理的CAL-27细胞凋亡率均高于空白对照组；琼脂糖凝胶电泳显示出典型的DNA梯形。结论：乳杆菌A-2代谢产物可以在体外抑制人舌鳞癌CAL-27细胞生长，诱导CAL-27细胞凋亡。
[Abstract]:Aim: to investigate the inhibitory effect of Lactobacillus A-2 metabolite on proliferation and apoptosis of human tongue squamous cell carcinoma (CAL-27) cells. Methods: Lactobacillus A-2 was cultured on restoration whey medium and restored whey inulin medium respectively to prepare metabolite 1 (LSPM1) and metabolite 2 (LSPM2). Cell morphology was observed by acridine orange staining, and apoptosis was detected by flow cytometry and DNA agarose gel electrophoresis. Results: in the 3.0mg/mL~48mg/mL region of LSPM1 and LSPM2, with the increase of the concentration of metabolites, the inhibitory effect on CAL-27 cells was gradually enhanced. The half inhibitory concentrations (IC50) of LSPM1 and LSPM2 were 15.1mg/mL and 10.27 mg / mL, respectively. The apoptotic cells showed irregular nuclei, crumpled cells, crescent or horseshoe shape, and poor adhesion. The results of flow cytometry showed that the apoptotic rate of CAL-27 cells treated with LSPM1 and LSPM2 was higher than that of the control group. Agarose gel electrophoresis showed a typical DNA trapezoid. Conclusion: Lactobacillus A-2 metabolites can inhibit the growth of human tongue squamous cell carcinoma (CAL-27) cells and induce apoptosis of CAL-27 cells in vitro.
【学位授予单位】：佳木斯大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R739.86

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