NNK对NCTC 1469细胞毒性作用的研究

doi:10.13560/j.cnki.biotech.bull.1985.2015.09.031

生物技术通报 ›› 2015, Vol. 31 ›› Issue (9): 218-223.doi: 10.13560/j.cnki.biotech.bull.1985.2015.09.031

NNK对NCTC 1469细胞毒性作用的研究

韩亚伟, 王西华, 陈利平, 时桂芹, 孙丽萍, 周文珊

(郑州轻工业学院食品与生物工程学院，郑州 450001)

收稿日期:2015-01-09 出版日期:2015-09-15 发布日期:2015-09-16
作者简介:王西华，男，硕士研究生，研究方向：发酵工程；E-mail：13673615603@163.com
基金资助:
郑州市科技局科技攻关项目(121PPTGG362-16)，河南省教育厅青年骨干教师项目(2012GG-117)

Toxic Effects of NNK on NCTC 1469 Cells

Han Yawei, Wang Xihua, Chen Liping, Shi Guiqin, Sun Liping, Zhou Wenshan

(College of Food and Bioengineering，Zhengzhou University of Light Industry，Zhengzhou 450001)

Received:2015-01-09 Published:2015-09-15 Online:2015-09-16

摘要/Abstract

摘要： 研究4-(甲基亚硝氨基)-1-(3-吡啶基)-1-丁酮(NNK)对小鼠肝细胞NCTC 1469细胞的毒性作用。利用MTT比色法检测5个不同NNK浓度(0.1、0.2、0.3、0.4、0.5 mg/mL)对细胞的毒性作用；倒置相差显微镜观察细胞的形态变化；流式细胞仪检测细胞凋亡率；实时荧光定量PCR检测凋亡基因Bcl-2和Bax的表达。MTT显示，NNK能降低NCTC 1469细胞存活率，并呈剂量和时间依赖性。镜检结果可见细胞皱缩，密度降低；流式细胞仪检测显示，0.1、0.2、0.3、0.4、0.5 mg/mL的NNK作用24 h后凋亡率分别为(16.79±2.01)%、(26.87±1.67)%、(41.78±3.19)%、(50.89±3.94)%和(65.86±4.54)%，显著高于对照组(7.46±1.58)%；荧光定量PCR结果显示，随着NNK浓度的增加，Bcl-2 mRNA的表达减少，Bax mRNA的表达逐渐上升。NNK能够降低NCTC 1469细胞存活率并诱导其凋亡，作用呈剂量和时间依赖性。

关键词: NNK, NCTC 1469细胞, 细胞凋亡, 流式细胞仪

Abstract: This work is to investigate the cytotoxicity in NCTC 1469 cells induced by 4-(methyInitrosamino)-1-(3-pyridyl)-1-butanone(NNK). MTT assay was used to analyze the toxic effects on NCTC 1469 cells under various concentrations(0.1, 0.2, 0.3, 0.4, and 0.5 mg/mL)of NNK at different times. Morphologic changes of treated NCTC 1469 cells with NNK were observed by the inverted microscopy. Flow cytometry(FCM)and real-time quantitative PCR were applied to measure the apoptosis rate and the expression of apoptotic gene Bcl-2 and Bax mRNA. NNK depressed the viability of NCTC 1469 cells with a pattern of the dose-dependent and time-dependent. Typical morphological changes of cells shrinkage and the decrease of density were observed by the inverted microscopy. FCW revealed that inhibited cell proliferation and induced death were of dose-dependent, i.e., apoptotic rates were(16.79 ± 2.01)%, (26.87 ± 1.67)%, (41.78 ± 3.19)%, (50.89 ± 3.94)% and(65.86 ± 4.54)% respectively at 24 h after the cells treated with the concentrations(0.1, 0.2, 0.3, 0.4, and 0.5 mg/mL)of NNK, showing much higher than the control of(7.46 ± 1.58)%. Real-time quantitative PCR showed that with the raising of NNH concentration, the expression of gene Bcl-2 decreased and the expression of gene Bax gradually increased. Conclusively, NNK could obviously abate the viability of NCTC 1469 cells and induce the apoptosis of NCTC 1469 cells in a pattern of dose-dependent and time-dependent.

Key words: NNK, NCTC 1469 cells, apoptosis, flow cytometry

韩亚伟, 王西华, 陈利平, 时桂芹, 孙丽萍, 周文珊. NNK对NCTC 1469细胞毒性作用的研究[J]. 生物技术通报, 2015, 31(9): 218-223.

Han Yawei, Wang Xihua, Chen Liping, Shi Guiqin, Sun Liping, Zhou Wenshan. Toxic Effects of NNK on NCTC 1469 Cells[J]. Biotechnology Bulletin, 2015, 31(9): 218-223.

参考文献

[1]毛友安. 卷烟烟气中烟草特有亚硝胺的新分析技术及其对酶活性影响的研究[R]. 湖南大学博士后研究工作报告. 长沙：2004.
[2]Rodgrman A. The composition of cigarette smoke：problems with lists of tumorigens[J]. Bitr Tabakforsch Int, 2003, 20：402-437.
[3]刘万峰, 王元英. 烟草中烟草特有亚硝胺的研究进展[J]. 中国烟草科学, 2002, 24(2)：11-14.
[4]Hecht SS. Biochemistry, biology, and carcinogenicity of tobacco-specific N-nirosamines[J]. Chem Res Toxicol, 1998, 11：559-603.
[5]Djordjevic MV, Brunnemann KD, Hoffmann D. Identification and analysis of a nicotine-derived N-nitrosamino acid and other nitrosamino acids in tobacco[J]. Carcinogenesis, 1989, 10(9)：1725-1731．
[6]World Health Organization technical series：No. 95 1. The scientific basis of tobacco product regulation[R]. Genewa：World Health Organization, 2008：69.
[7]谢剑平, 刘惠民, 朱茂祥, 等. 卷烟烟气危害性指数研究[J]. 烟草科技, 2009(2)：5-15．
[8]International Agency for Research on Cancer(IARC). IARC monographs on the evaluation of carcinogenic risks to humans：V01. 89. Smokeless tobacco and some tobacco-specific N-nitrosamines[M]. Lyon：International Agency for Research on Cancer, 2007：59.
[9]宁维, 陈利平, 李瑜, 等. 卷烟烟气体内安全评价方法研究[J]. 河南农业科学, 2013, 42(9)：152-156.
[10]宁维, 陈利平, 李瑜, 等. 全烟气致小鼠肺损伤模型的建立[J]. 烟草科技, 2013, 11：36-40.
[11]邹小农. 中国肺癌流行病学[J]. 中华肺癌防治杂志, 2007, 14(12)：881-883.
[12]Wynder EL, Hoffmann D. Smoking and lung cancer：scientific challenges and opportunities[J]. Cancer Res, 1994, 54(20)：5284-5295.
[13]Wu X, Zhao H, Suk R, et al. Genetic susceptibility to tobacco related cancer[J]. Oncogene, 2004, 23(38)：6500-6523.
[14]Akopyan G, Bonavida B. Understanding tobacco smoke carcinogen NNK and lung tumorigen- esis[J]. Int J Oncol, 2006, 29(4)：745-752.
[15]Haussmann HJ. Smoking and lung cancer：future research directions[J]. Int J Toxicol, 2007, 26(4)：353-364.
[16]Zhang H, Yang J Y, Zhou F, et al. Seed Oil of Brucea javanica induces apopotic death of acute myeloid leukemia cells via both the death receptors and the mitochondrial-related pathways[J]. Evid Based Complement Alternat Med, 2011, 28：965-969.
[17]Tsukaharas S, Yamamoto S, Shew TT, et al. Inhalation of low level formaldehyde increases the bcl 2/bax expression ratio in the hippocampus of immunologically sensitized mice[J]. Neuroimmunomodulation, 2006, 13(2)：63-68．
[18]Parikh N, Koshy C, Dhayabaran V, et al. The N-terminus and alpha-5, alpha-6 helices of the pro-apoptotic protein Bax, modulate functional interactions with the anti-apoptotic protein Bcl-xL[J]. BMC Cell Biol, 2007, 8：16-20.

NNK对NCTC 1469细胞毒性作用的研究

Toxic Effects of NNK on NCTC 1469 Cells

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价

[1]	米粱波, 曾伟主, 黄科学, 王得明, 堵国成, 周景文, 陈坚. 高通量筛选地衣芽孢杆菌DW2高产杆菌肽诱变菌株[J]. 生物技术通报, 2020, 36(7): 90-96.
[2]	尹晓梦, 曹雪玮, 王富军, 赵健, 张惠展. 雷公藤红素与凋亡蛋白突变体通过强化Nur77诱发凋亡通路发挥协同抗肿瘤作用[J]. 生物技术通报, 2020, 36(7): 119-129.
[3]	朱平, 杜力杰, 孟昆, 薛娟, 杨瑾, 李姗. 三型分泌系统效应蛋白调控细胞凋亡和焦亡的研究进展[J]. 生物技术通报, 2019, 35(4): 178-187.
[4]	胡建燃, 李平, 铁军, 金山. 紫丁香花精油的抗氧化和抗肿瘤活性研究[J]. 生物技术通报, 2019, 35(12): 16-23.
[5]	翟逸舟 ,卢美雅 ,赵健 ,王富军. 白树毒素融合蛋白的筛选及其抗肿瘤作用和凋亡途径研究[J]. 生物技术通报, 2018, 34(6): 204-212.
[6]	郭红艳, 高涵, 吴琦, 孙晓杰, 刘秀财, 赵立群. SGK3基因RNAi慢病毒载体的构建及其对乳腺癌MB-474细胞增殖和凋亡的影响[J]. 生物技术通报, 2018, 34(1): 247-252.
[7]	邓雨青, 李平, 周彦, 熊克才, 李中安. 植物细胞程序性死亡检测技术研究进展[J]. 生物技术通报, 2017, 33(3): 52-57.
[8]	侯欢欢, 卢佳, 章纬菁, 黄芳任文华. 少棘蜈蚣抗菌肽scolopin 2的克隆、表达与抗癌机制研究[J]. 生物技术通报, 2016, 32(9): 172-178.
[9]	齐仁立, 黄金秀, 龙定彪, 黄萍. MicroRNA与NF-κB调控的细胞凋亡[J]. 生物技术通报, 2015, 31(5): 27-31.
[10]	刘凤霞,刘文娟,李建勇,陈胜国. ERK1/2在食管鳞状细胞癌中促进细胞增殖、抑制凋亡及其调控机制的研究[J]. 生物技术通报, 2015, 31(10): 242-248.
[11]	齐仁立, 黄金秀, 杨飞云, 刘作华. 细胞凋亡中MicroRNA的调控作用[J]. 生物技术通报, 2014, 0(7): 20-25.
[12]	齐仁立,黄金秀,宋凡,陈英,黄萍. 共轭亚油酸诱导多种细胞凋亡[J]. 生物技术通报, 2014, 0(1): 53-57.
[13]	和四梅, 张丽莉, 王艺磊, 王国栋. Caspase细胞凋亡通路及其在水生无脊椎动物的研究进展[J]. 生物技术通报, 2013, 0(9): 54-61.
[14]	郑娟娟, 张雨, 许文涛, 黄昆仑. 锌对哺乳动物细胞损伤的保护机制[J]. 生物技术通报, 2013, 0(4): 21-26.
[15]	吴薇,刘朝奇. 钙网蛋白通过内质网应激介导细胞凋亡的分子机制[J]. 生物技术通报, 2013, 0(10): 24-27.