《Journal of Oral and Maxillofacial Surgery》 ›› 2014, Vol. 24 ›› Issue (5): 336-. doi: 10.3969/j.issn.1005-4979.2014.05.003

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

SUN Zhi-hui, WANG Xing-yu, GUAN Hong, MENG Cun-fang, GUAN Jian   

  1. 1. Department of Oral and Maxillofacial Surgery, Hospital of Stomatology, Jiamusi University, Jiamusi 154004; 2. Research Center of Microecological Engineering Technology, Qiqihaer 161000, Heilongjiang Province, China
  • Online:2014-10-28 Published:2015-02-10

乳杆菌A2代谢产物对人舌鳞癌细胞系Cal-27凋亡的诱导

孙智辉,王心彧,关宏,孟存芳,关键   

  1. 1. 佳木斯大学附属口腔医院颌面外科,黑龙江   佳木斯   154004;
    2. 齐齐哈尔医学院微生态工程技术研究中心,黑龙江  齐齐哈尔   161000
  • 通讯作者: 关键,主任医师. E-mail:guanjian1207@163.com
  • 作者简介:孙智辉,男,(1987—),硕士研究生.
  • 基金资助:

    黑龙江省自然科学基金面上项目(H201357)

Abstract: 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.

Key words:  Lactobacillus sp. A2′s metabolites, whey protein, human tongue squamous carcinoma cells, cells apoptosis

摘要: 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。

关键词:  , 乳杆菌A2代谢产物;乳清蛋白;人舌鳞癌细胞;细胞凋亡

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