三氯卡班诱导小鼠乳腺上皮细胞癌变的研究

发布时间：2018-01-24 16:32

本文关键词： 三氯卡班乳腺上皮细胞致癌活性氧簇 DNA损伤　出处：《南京农业大学学报》2017年05期 　论文类型：期刊论文

【摘要】：[目的]广谱抗菌剂三氯卡班(TCC)在各种环境介质和生物体中被频繁检出。为了研究TCC对动物的危害,观察了TCC对正常小鼠乳腺上皮细胞(Ep H4-Ev)的急慢性损伤作用,评价其对乳腺上皮细胞的诱癌作用,为防止TCC在动物用品中的滥用提供理论依据。[方法]采用MTT法确定TCC的最佳工作浓度后,作用于Ep H4-Ev细胞,以48 h为1个药物作用循环,分别作用3和6个循环(命名为T3和T6细胞)后,使用MTT法检测细胞增殖,流式细胞术检测细胞内活性氧簇(ROS)水平,Western blot法测定细胞内磷酸化组蛋白(p-H2AX)的蛋白表达量,软琼脂克隆试验检测细胞非停泊性生长能力,细胞划痕试验测定细胞迁移能力,用此指标作为检测TCC引起细胞癌变的指标。[结果]Ep H4-Ev细胞在0.195μmol·L-1TCC作用下,细胞活性最大,故0.195μmol·L-1为TCC的工作浓度;T3和T6细胞的细胞增殖水平、ROS水平、p-H2AX蛋白表达量及软琼脂上的细胞集落个数均显著高于细胞对照;T6细胞的细胞划痕之间的距离显著小于细胞对照。[结论]TCC能够诱导正常的Ep H4-Ev细胞发生癌变。
[Abstract]:[Objective] TCCs, a broad-spectrum antibacterial agent, were frequently detected in various environmental media and organisms. In order to study the harm of TCC to animals. To observe the acute and chronic injury effect of TCC on mammary epithelial cells of normal mice, and to evaluate its carcinogenic effect on mammary epithelial cells. In order to prevent TCC abuse in animal supplies to provide theoretical basis. [Methods] the optimal working concentration of TCC was determined by MTT method, and EpH4-Ev cells were treated with TCC for 48 h. After 3 and 6 cycles (named T3 and T6 cells), cell proliferation was detected by MTT assay, and intracellular reactive oxygen species (Ros) levels were detected by flow cytometry. The protein expression of phosphorylated histone (p-H2AX) was measured by Western blot assay, and the ability of non-moored growth was detected by soft Agar clone assay. The cell migration ability was measured by cell scratch test, which was used as a marker for the detection of cell carcinogenesis induced by TCC. [Results: the activity of EpH4-Ev cells was the highest under the treatment of 0.195 渭 mol 路L -1 TCC, so 0.195 渭 mol 路L -1 was the working concentration of TCC. The cell proliferation level of T3 and T6 cells and the expression of p-H2AX protein at Ros level and the number of colony on soft Agar were significantly higher than those in the control group. The distance between scratch of T 6 cells was significantly smaller than that of control cells. [Conclusion: TCC can induce carcinogenesis of normal EpH4-Ev cells.
【作者单位】：南京农业大学中兽医学研究室;山东省平原县畜牧兽医局;
【基金】：国家自然科学基金项目(31572557) 国家公益性行业(农业)科研专项(2013030404,201403051) 江苏高校优势学科建设工程项目(PAPD)
【分类号】：S852.3
【正文快照】： (1.Institute of Traditional Chinese Veterinary Medicine,Nanjing Agricultural University,Nanjing 210095,China;2.Animal Husbandry and Veterinary Bureau of Pingyuan Country,Shandong Province,Dezhou 253100,China)三氯卡班(triclocarban,TCC),其化学名称为3,4,4-

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