乳杆菌A2代谢产物对人舌鳞癌细胞系Cal-27凋亡的诱导

Lactobacillus sp. A2′s Metabolites Induces Apoptosis in Human Tongue Cancer Cal-27 Cells

目的：观察乳杆菌A2代谢产物LM1和LM2对人口腔舌鳞癌细胞系Cal-27细胞增殖抑制和诱导凋亡作用。方法：将乳杆菌A2经复原乳清培养基培养制备相应的代谢产物1（LM1）,除去代谢产物中的钙离子得到产物2（LM2）;用MTT法和流式细胞仪检测口腔舌鳞状细胞癌的细胞凋亡的情况;琼脂糖凝胶电泳检测DNA片段的变化。结果：乳杆菌A2代谢产物LM1和LM2作用Cal-27细胞48 h时,最大抑制率分别为（39.9±1.56）%和（30.5±6.85）%;Annexinv-FITC/PI染色法检测LM2（8 mg/mL）培养40、44 h时,细胞早期凋亡率分别为19.93%和21.42%;琼脂糖凝胶电泳LM2（8 mg/mL）培养54 h时,在250~750 bp之间显现出典型的＂梯＂状DNA条带。结论：LM1和LM2能抑制口腔舌鳞癌细胞Cal-27的增殖,呈剂量依赖性关系,LM2能够诱导细胞凋亡。

Objective： To investigate the effects of Lactobacillus sp. A2′s metabolites on the proliferation and apoptosis of tongue squamous cell caricoma Cal-27 cells. Method： The recovery of whey Lactobacillus medium（whey） was used to prepare the corresponding metabolites of culture 1（LM1）, and being removed the calcium ions metabolites to give the product 2（LM2）. MTT assay was used to test the inhibition effect of LM2 on Cal-27 cells. AnnexinV-FITC/PI staining flow cytometry was used for cell apoptosis. Agarose gel electrophoresis was used to detect the DNA fragments in Cal-27 cells after the treatment by LM2. Results： After Cal-27 cells were treated with LM1 or LM2 for 48 h, maximum inhibition rates were（39.9%±1.56%） and（30.5%±6.85%） respectively. Flow cytometry analysis with annexinV-FITC/PI showed that after treated with LM2（8 mg/mL） for 40 h and 44 h, cell apoptosis rate was 19.93% and 21.42%. The typical DNA ladders of the treated cells were detected by agarose gel electrophoresis,when cultured by LM2（8 mg/mL） for 54 h, the DNA ladders were between 250~750 bp. Conclusion： LM2 may inhibit the proliferation and induce the apoptosis of Cal-27 cells in a dose-dependent manner.