本文關(guān)鍵詞： 三氯卡班 乳腺上皮細(xì)胞 致癌 活性氧簇 DNA損傷　出處：《南京農(nóng)業(yè)大學(xué)學(xué)報(bào)》2017年05期 　論文類型：期刊論文

【摘要】：[目的]廣譜抗菌劑三氯卡班(TCC)在各種環(huán)境介質(zhì)和生物體中被頻繁檢出。為了研究TCC對(duì)動(dòng)物的危害,觀察了TCC對(duì)正常小鼠乳腺上皮細(xì)胞(Ep H4-Ev)的急慢性損傷作用,評(píng)價(jià)其對(duì)乳腺上皮細(xì)胞的誘癌作用,為防止TCC在動(dòng)物用品中的濫用提供理論依據(jù)。[方法]采用MTT法確定TCC的最佳工作濃度后,作用于Ep H4-Ev細(xì)胞,以48 h為1個(gè)藥物作用循環(huán),分別作用3和6個(gè)循環(huán)(命名為T(mén)3和T6細(xì)胞)后,使用MTT法檢測(cè)細(xì)胞增殖,流式細(xì)胞術(shù)檢測(cè)細(xì)胞內(nèi)活性氧簇(ROS)水平,Western blot法測(cè)定細(xì)胞內(nèi)磷酸化組蛋白(p-H2AX)的蛋白表達(dá)量,軟瓊脂克隆試驗(yàn)檢測(cè)細(xì)胞非停泊性生長(zhǎng)能力,細(xì)胞劃痕試驗(yàn)測(cè)定細(xì)胞遷移能力,用此指標(biāo)作為檢測(cè)TCC引起細(xì)胞癌變的指標(biāo)。[結(jié)果]Ep H4-Ev細(xì)胞在0.195μmol·L-1TCC作用下,細(xì)胞活性最大,故0.195μmol·L-1為T(mén)CC的工作濃度;T3和T6細(xì)胞的細(xì)胞增殖水平、ROS水平、p-H2AX蛋白表達(dá)量及軟瓊脂上的細(xì)胞集落個(gè)數(shù)均顯著高于細(xì)胞對(duì)照;T6細(xì)胞的細(xì)胞劃痕之間的距離顯著小于細(xì)胞對(duì)照。[結(jié)論]TCC能夠誘導(dǎo)正常的Ep H4-Ev細(xì)胞發(fā)生癌變。
[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.
【作者單位】： 南京農(nóng)業(yè)大學(xué)中獸醫(yī)學(xué)研究室;山東省平原縣畜牧獸醫(yī)局;
【基金】：國(guó)家自然科學(xué)基金項(xiàng)目(31572557) 國(guó)家公益性行業(yè)(農(nóng)業(yè))科研專項(xiàng)(2013030404,201403051) 江蘇高校優(yōu)勢(shì)學(xué)科建設(shè)工程項(xiàng)目(PAPD)
【分類號(hào)】：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),其化學(xué)名稱為3,4,4-

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