华西口腔医学杂志

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嗜酸乳杆菌对人舌癌细胞增殖的影响

史晓艳 陈骏 车团结 白德成 何祥一   

  1. 1.兰州大学口腔医院修复科; 2.兰州大学生命科学院细胞与分子生物学实验室;3.兰州大学基础医学院甘肃省新药临床前研究重点实验室, 兰州730000
  • 收稿日期:2012-02-25 修回日期:2012-02-25 出版日期:2012-02-01 发布日期:2012-02-01
  • 通讯作者: 何祥一,Tel:13038730686
  • 作者简介:史晓艳(1983—),女,山东人,硕士
  • 基金资助:

    甘肃省科技攻关计划基金资助项目(0709TCYA053)

A study on human tongue cancer cells’proliferation affected by Lactobacillus acidophilus

Shi Xiaoyan1, Chen Jun1, Che Tuanjie2, Bai Decheng3, He Xiangyi1   

  1. 1. Dept. of Prosthodontics, School of Stomatology, Lanzhou University, Lanzhou 730000, China; 2. Laboratory of Cell and Molecular Biology, School of Life Science, Lanzhou University, Lanzhou 730000, China; 3. Key Laboratory of Preclinical Study for New Drugs of Gansu Province, Basic Medical College, Lanzhou University, Lanzhou 730000, China
  • Received:2012-02-25 Revised:2012-02-25 Online:2012-02-01 Published:2012-02-01
  • Contact: He Xiangyi,Tel:13038730686
  • About author:Shi Xiaoyan(1983—),女,山东人,硕士

摘要:

目的研究嗜酸乳杆菌对人舌癌细胞Tca8113增殖及细胞周期的影响。方法体外培养Tca8113细胞,分别将不同稀释度(原液和4、16倍稀释)的嗜酸乳杆菌上清液、灭活菌液和无细胞提取物与Tca8113细胞共培养,采用倒置显微镜观察细胞形态并行细胞计数,磺酰罗丹明B(SRB)法测定细胞增殖率,流式细胞术分析嗜酸乳杆菌各组分对Tca8113细胞增殖及细胞周期的影响,激光扫描共聚焦显微镜(CLSM)检测细胞内自由基和Ca2+含量。结果嗜酸乳杆菌各组分作用于Tca8113细胞48 h后,在倒置显微镜下观察,细胞由菱形、多角形、铺路石状变为细长形。细胞计数与SRB实验分析:在不同稀释度同一培养时间与不同培养时间同一稀释度培养条件下,嗜酸乳杆菌各组分均可明显抑制Tca8113细胞增殖,抑制力随稀释度增加而降低,随培养时间延长而增强。流式细胞术分析:嗜酸乳杆菌各组分作用Tca8113细胞48 h后,细胞增殖指数降低(P<0.01)。CLSM检测:嗜酸乳杆菌各组分作用Tca8113细胞48 h后细胞内自由基和Ca2+含量均升高(P<0.01)。结论嗜酸乳杆菌代谢产物、灭活菌液、无细胞提取物均可抑制Tca8113细胞增殖,可能与菌体及其代谢产物引起细胞内自由基含量增多、Ca2+超载有关。

关键词: Tca8113细胞, 嗜酸乳杆菌, 细胞增殖, 激光扫描共聚焦显微镜

Abstract:

Objective To study the effects of Lactobacillus acidophilus(L.acidophilus) on the proliferation and cell  cycle distribution of human tongue cancer cells(Tca8113 cells). Methods In vitro cultivated human Tca8113 cells were treated by L.acidophilus supernatant, inactivated bacilli, cell free extracts and normal culture medium respectively, which were 1, 4, 16-fold(s) dilutelly, to investigate the proliferous effects of Tca8113 cells using of inverted microscope, cell counting, sulforhodamine B(SRB) and flow cytometry. The free radicals and Ca2+ in Tca8113 cells were also studied by confocal laser scanning microscope(CLSM). Results At the 48th hour after adding different L. acidophilus components, the Tca8113 cells changed in shape from the diamond-like, polygonal and slabs into the elongated form. In the condition of different times and different culture concentrations, the proliferation of Tca8113 cells was significantly inhibited by L.acidophilus components, which enhanced as the time prolonged and the concentrations of each L. acidophilus components increased according to the cell counting and the SRB experimental analysis. The cell proliferation index(CPI) was significantly reduced(P<0.01). The free radicals and Ca2+ in Tca8113 cells under the effect of each L. acidophilus components for 48 h indicated an obviously rising(P<0.01). Conclusion L. acidophilus restrains the proliferation of Tca8113 cells, which might be due to the increase in quantity of free radicals and Ca2+ in Tca8113 cells, and might be resulted from the release of metabolic products of L.acidophilus.

Key words: Tca8113 cell, Lactobacillus acidophilus, cell proliferation, confocal laser scanning microscope