新城疫病毒HN基因诱导人肺癌细胞SPC-A1凋亡的作用机制

PDF(538 KB)

中国生物化学与分子生物学报 ›› 2006, Vol. 22 ›› Issue (03) : 222-222~227.

研究论文

新城疫病毒HN基因诱导人肺癌细胞SPC-A1凋亡的作用机制

连海^1,2);金宁一^1)*;李霄¹⁾;陈立刚¹⁾;张静敏¹⁾;管国芳¹⁾;孙丽丽¹⁾;李雪梅¹⁾;郑洪玲¹⁾;崔英姬³⁾

作者信息 +

Mechanisms of Apoptosis in Human Lung Carcinoma Cells SPC-A1 Induced with Hemagglutininneuraminidase(HN) Gene from Newcastle Disease Virus

LIAN Hai^１）;2) ;JIN Ning-Ｙi ^1）* ;LI Xiao ^1）;CHEN Li-Ｇang ^1）;ZHANG Jing-Ｍin ^1）;GUAN Guo-Ｆang ^1）;SUN Li-Ｌi ^1）;LI Xue-Ｍei ^1）;ZHENG Hong-Ｌing ^1）;CUI Ying-Ji ³⁾

Author information +

文章历史 +

摘要

为了探索新城疫病毒血凝素神经氨酸酶(HN)基因对人肺癌细胞SPC-A1的作用及机制，将含有HN基因的重组质粒pVHN经脂质体介导转染人肺癌细胞SPC-A1，通过MTT方法检测细胞活力；采用吖啶橙/溴化乙锭（AO/EB）染色分析肿瘤细胞凋亡；罗丹明123和DCFA染色测定线粒体跨膜电位（ΔΨm）和活性氧水平变化；流式细胞仪分析MHCⅠ分子表达；底物染色反应检测caspase-3活性结果重组质粒pVHN转染人肺癌细胞SPCA1 48 h后，细胞活力明显降低； AO/EB染色可见明显的细胞凋亡形态学变化；与空质粒对照相比，线粒体ΔΨm下降(Ｐ<0.01)，活性氧水平升高(Ｐ<0.05)；细胞表面MHC-Ⅰ分子表达上调(P>0.05)；caspase-3活性增强(Ｐ<0.01). 以上结果提示，新城疫病毒HN基因能够上调SPC-A1细胞表面MHC-Ⅰ分子表达，并通过上调ROS水平，下调线粒体ΔΨm，进而激活caspase-3，最终诱导人肺癌细胞凋亡.

Abstract

To investigate the effect of hemagglutinin-neuraminidase(HN) gene from Newcastle disease virus on human lung carcinoma cells SPC-A1, the recombinant plasmid pVHN was introduced into human lung carcinoma cells SPC-A1 byliposome-mediated transfection. The viability of SPC-A1 cells was determined by MTT staining and apoptosis was detected by AO/EB staining. The alteration of mitochondrial transmembrane potential and ROS level of the cells was detected by flow cytometry (FCM) with rhodamine 123 and DCFA staining.The expression of MHC-Ⅰ was detected by FCM and the activation of caspase-3 was assayed by its substrate color reaction. Viability of SPC-A1 cells was decreased significantly and recombinant plasmid could lead to obviously morphological apoptotic changes of SPC-A1 cells by introduction of pVHN into SPC-A1 cells for 48 hours. Mitochondrial transmembrane potential was decreased(Ｐ<0.01) , ROS level of the cells was increased (Ｐ<0.05), theexpression of MHC-Ⅰ(P>0.05) and caspase-3 activity (Ｐ<0.01)was increased, compared with control cells.The above results showed increase of MHC-Ⅰ expression andsignificant apoptosis in SPC-A1 cells can be induced by recombinant plasmid pVHN. Apoptosis may be resulted from the increase of ROS level, the decrease of mitochondrial transmembrane potential and activation of Caspase-3.

导出引用

连海,;金宁一;李霄;陈立刚;张静敏;管国芳;孙丽丽;李雪梅;郑洪玲;崔英姬. 新城疫病毒HN基因诱导人肺癌细胞SPC-A1凋亡的作用机制[J]. 中国生物化学与分子生物学报, 2006, 22(03): 222-222~227

LIANHai;;JINNing-Ｙi;LIXiao;CHENLi-Ｇang;ZHANGJing-Ｍin;GUANGuo-Ｆang;SUNLi-Ｌi;LIXue-Ｍei;ZHENGHong-Ｌing;CUIYing-Ji. Mechanisms of Apoptosis in Human Lung Carcinoma Cells SPC-A1 Induced with Hemagglutininneuraminidase(HN) Gene from Newcastle Disease Virus[J]. Chinese Journal of Biochemistry and Molecular Biology, 2006, 22(03): 222-222~227

中图分类号： Q255 Q75

参考文献

1 吴一龙, 张明和, 廖美琳，周清华，蒋国梁，支修益，王俊，陈刚，王思愚，杨学宁，孙燕. 肺癌的诊断和分期临床指引. 中国肺癌杂志(Wu YiLong,Zhang MingHe,Liao MeiLin,Zhou QingHua,Jiang GuoLiang,Zhi XiuYi,Wang Jun,Chen Gang,Wang SiYu,Yang XueNing,Sun Yan. Diagnosis and clinical staging of lung cancer. Chin J Lung Cancer), 2003, 6 (5) : 330～334
2 Schirrmacher V, Griesbach A, Ahlert T. Antitumor effects of Newcastle Disease Virus in vivo: local versus systemic effects. Int J Oncol,2001,18(5)：945～952
3 Phuangsab A, Lorence R　M, Reichard K　W, Peeples M　E, Walter R　J.Newcastle disease virus therapy of human tumor xenografts: antitumor effects of local or systemic administration. Cancer Lett,2001,172(1)：27～36
4 Zorn U, Duensing S, Langkopf F, Anastassiou G, Kirchner H, Hadam M, KnuverHopf J, Atzpodien J. Active specific immunotherapy of renal cell carcinoma: cellular and humoral immune responses. Cancer Biother Radiopharm, 1997,12(3)：157～165
5 Zeng J, Fournier P, Schirrmacher V. Induction of interferon-alpha and tumor necrosis factor related apoptosis-inducing ligand in human blood mononuclear cells by hemagglutinin-neuraminidase but not F protein of Newcastle disease virus.Virology， 2002,297： 19～30
6 龚伟，薛丽娟，孙大辉，王宏伟，葛涛，罗琴芳，李萍，金宁一. pIRVVP3/pIRHNVP3核酸表达质粒的构建及对肿瘤细胞的影响.免疫学杂志(Gong Wei, Xue LiＪuan, Sun DaＨui,Wang HongＷei,Ge Tao, Luo QinＦang,Li Ping,Jin NingＹi. Construction of nucleic acid expressing vaccines of pIRVP3/pIRHNVP3 and its effects on the tumor cells.Immunol J), 2002,18(1):30～32
7 米志强，金宁一，龚伟，薛立娟，连海，解丽华，李萍.pVVP3和pVHN核酸疫苗的构建、表达及对肿瘤细胞的影响.中国生物化学与分子生物学报(Mi ZhiQiang,Jin NingYi,Gong Wei,Xue LiJuan,Lian Hai, Xie LiHua, Li Ping. Construction and expression of the nucleic acid vaccine pVVP3 and pVHN and its effect on tumor cell. Chin J Biochem Mol Biol),2003,19(6):704～708
8米志强, 金宁一, 龚伟, 薛立娟, 孙大辉, 葛涛, 连海, 解丽华. 新城疫病毒HN 基因构建的核酸疫苗抗肿瘤作用研究. 中国肿瘤生物治疗杂志,(Mi ZhiＱiang, Jin NingYi, Gong Wei, Xue LiJuan, Sun DaHui, Ge Tao, Lian Hai, Xie LiHua. The antitumor effect of the nucleic vaccine constructed with newcastle disease virus HN gene. Chin J Cancer Biother), 2003,10 (2): 93～96
9 殷震,刘景华主编,动物病毒学.第2版.北京：科学出版社(Yin Zhen ,Liu JingＨua ed . Animal Virology,2nd ed. Beijing:Science Press) , 1997： 351～352
10 Yip D, Strickland A　H, Karapetis C　S, Hawkins C　A, Harper P　G. Immunomodulation therapy in colorectal carcinoma. Cancer Treat Rev, 2000，26: 169～190
11 Pecora A　L, Rizvi N, Cohen G　I, Meropol N　J, Sterman D, Marshall J　L, Goldberg S, Gross P, O′Neil J　D, Groene W　S, Roberts M　S, Rabin H, Bamat M　K, Lorence R　M.Phase I trial of intravenous administration of PV701, an oncolytic virus in patients with advanced solid cancers. J Clin Oncol, 2002， 20: 2251～2266
12 Batliwalla F　M, Bateman B　A, Serrano D, Murray D, Macphail S, Maino V　C, Ansel J　C, Gregersen P　K, Armstrong C　A. A 15-year follow-up of AJCC stage III malignant melanoma patients treated postsurgically with Newcastle disease virus (NDV) oncolysate and determination of alterations in the CD8 T cell repertoire. Mol Med, 1998， 4: 783～794
13 Hu W, Kavanagh J　J. Anticancer therapy targeting the apoptotic pathway.Lancet Oncol, 2003,4(12):721～729
14 Zamzami N, Marchetti P, Castedo M, Decaudin D, Macho A, Hirsch T, Susin S　A, Petit P　X, Mignotte B, Kroemer G. Sequential reduction of mitochondrial transmembrane potential and generation of reactive oxygen species in early programmed cell death. J Exp Med. 1995,182(2):367～377
15 Jacobson M　D, Raff M　C. Programmed cell death and Bcl-2 protection in very low oxygen. Nature. 1995,374(6525):814～816
16 Thornberry N A, Lazebnik Y. Caspases: enemies within. Science 1998, 281(5381):1312～1316

PDF(538 KB)

Accesses

Citation

Detail

段落导航

摘要
Abstract
关键词
Key words
引用本文
参考文献

收稿日期	修回日期	出版日期
2005-07-15	1900-01-01	2006-03-20
发布日期
2006-03-20

选择文件类型/文献管理软件名称

选择包含的内容

摘要

Abstract

关键词

Key words

引用本文

{{custom_sec.title}}

{{custom_sec.title}}

参考文献

{{custom_fnGroup.title_cn}}

脚注

模态框（Modal）标题

选择文件类型/文献管理软件名称

选择包含的内容

摘要

Abstract

关键词

Key words

引用本文

{{custom_sec.title}}

{{custom_sec.title}}

参考文献

{{custom_fnGroup.title_cn}}

脚注