[1]李宁,陈伟,张亚飞黎川,等.抗肿瘤转移肝素酶特异性抗体的筛选及鉴定[J].第三军医大学学报,2011,33(08):771-775.
 Li Ning,Chen Wei,Zhang Yafei,et al.Screening and identification of specific anti-heparanase monoclonal antibody inhibiting tumor metastasis[J].J Third Mil Med Univ,2011,33(08):771-775.
点击复制

抗肿瘤转移肝素酶特异性抗体的筛选及鉴定(/HTML )
分享到:

《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
33卷
期数:
2011年第08期
页码:
771-775
栏目:
论著
出版日期:
2011-04-30

文章信息/Info

Title:
Screening and identification of specific anti-heparanase monoclonal antibody inhibiting tumor metastasis
作者:
李宁陈伟张亚飞黎川 吴玉云 汪苏敏 房殿春 杨仕明
第三军医大学西南医院:全军消化病研究所,放射科
Author(s):
Li Ning Chen Wei Zhang Yafei Li Chuan Wu Yuyun Wang Shumin Fang Dianchun Yang Shiming
Institute of Gastroenterology;Department of Radiology, Southwest Hospital, Third Military Medical University, Chongqing, 400038, China
关键词:
肝素酶单克隆抗体抗肿瘤转移
Keywords:
heparanase monoclonal antibody tumor metastasis inhibition
分类号:
R392-33;R392.11;R73-362
文献标志码:
A
摘要:
目的    通过鉴定制备的人肝素酶单克隆抗体,筛选出具有抗肿瘤转移特异性的单克隆抗体株,并大量制备。    方法    采用Western blot技术,以HepG2、SGC-7901、MKN45、SW480、U2OS、MCF-7细胞总蛋白为样本,对已制备的3株人肝素酶单克隆抗体进行初步鉴定,筛选出能够与肝素酶蛋白特异性结合的抗体,并采用免疫细胞化学技术进一步验证筛选出的抗体特异性,然后通过Transwell体外侵袭实验观察筛选出的肝素酶单克隆抗体对肿瘤转移的抑制作用。    结果    已制备的3株人肝素酶单克隆抗体均能与不同肿瘤细胞肝素酶蛋白结合,其中12号抗体与商品化肝素酶单克隆抗体在MCF-7细胞中均检测出肝素酶弱阳性表达,其特异性最佳;免疫细胞化学结果也显示在SGC-7901、HepG2、MKN45、SW480和U2OS细胞中12号抗体能检测出肝素酶阳性表达,而在MCF-7细胞中检测出肝素酶弱阳性表达;Transwell体外侵袭实验结果显示12号抗体(100 μg,浓度约1 mg/ml)作用于SGC-7901、HepG2、MKN45、SW480和U2OS细胞48 h后,其穿膜细胞数与正常小鼠IgG对照组相比明显减少(P<0.05)。    结论    纯化制备的人肝素酶单克隆抗体能够与肿瘤细胞表达的肝素酶蛋白特异性结合,并通过抑制肝素酶蛋白达到抑制肿瘤细胞的扩散与转移。
Abstract:
Objective    To identify monoclonal antibodies against human heparanase and to screen a monoclonal antibody specifically inhibiting tumor metastasis.     Methods    Through Western blotting, three self-prepared monoclonal antibodies (No. 7, No. 11, and No. 12) were subjected to heparanase specificity identification and screening with total proteins of HepG2, SGC-7901, MKN45, SW480, U2OS and MCF-7 cells as samples. Immunocytochemical assay was used to further verify the specificity of the screened antibody. Transwell cell invasion assay in vitro was used to observe the inhibitory effect of the screened antibody on tumor metastasis.     Results    All the three monoclonal antibodies could bind heparanase in different tumor cells. Monoclonal antibody No. 12, as well as a commercial anti-heparanase monoclonal antibody as positive control, could detect weak positive expression of heparanase in MCF-7 cells, indicating the best specificity. Immunocytochemical assay results showed that monoclonal antibody No. 12 could detect positive heparanase expression in SGC-7901, HepG2, MKN45, SW480 and U2OS cells, and weak positive heparanase expression in MCF-7 cells. Transwell cell invasion assay results demonstrated that the migrated cell numbers of SGC-7901, HepG2, MKN45, SW480 and U2OS cells treated by monoclonal antibody No. 12 (100 μg, 1 mg/ml approximately) for 48 h were significantly lowered as compared with those treated by normal mouse IgG for 48 h (P<0.05).     Conclusion    A self-prepared monoclonal antibody can specifically bind heparanase in tumor cells and inhibit the invasion and metastasis of tumor cells by suppressing heparanase activity.

相似文献/References:

[1]左国华,梁平,李洪艳.肝细胞癌免疫磁珠的制备及鉴定[J].第三军医大学学报,2005,27(16):1700.
[2]陈曦,黄悦,于彬,等.结直肠癌单克隆抗体的制备及生物学特性鉴定[J].第三军医大学学报,2007,29(10):896.
 CHEN Xi,HUANG Yue,YU Bin,et al.Preparing and identifying a monoclonal antibody against human colorectal carcinoma[J].J Third Mil Med Univ,2007,29(08):896.
[3]罗萍,陈洪章,曾明,等.肠出血性大肠杆菌O157∶H7志贺毒素ⅡA1亚单位单克隆抗体的制备和生物学特性鉴定[J].第三军医大学学报,2008,30(08):698.
 LUO Ping,CHEN Hong-zhang,ZENG Ming,et al.Preparation and biological characterization of monoclonal antibody against shiga toxin Ⅱ A1 subunit of enterohemorrhagic E.coli O157:H7[J].J Third Mil Med Univ,2008,30(08):698.
[4]黄志,徐秀娟.nAChR单克隆抗体建立重症肌无力被动转移小鼠模型的研究[J].第三军医大学学报,2006,28(13):1397.
[5]邹佳,葛晓冬,刘友生,等.人源脂多糖结合蛋白单克隆抗体的筛选及初步鉴定[J].第三军医大学学报,2006,28(10):1032.
[6]孙仁美,王鲁,宋秋荷,等.单克隆抗体Mab03.2C1-C2靶抗原性质的初步研究[J].第三军医大学学报,2006,28(09):976.
[7]何凤田,李蓉芬,郑英如,等.噬菌体抗体库技术制备胃癌单抗MGd1的抗独特型抗体[J].第三军医大学学报,2005,27(19):1911.
[8]宋秋荷,王鲁,郝飞,等.酵母菌烯醇化酶单克隆抗体的制备及鉴定[J].第三军医大学学报,2005,27(13):1329.
[9]唐玮,王鲁,唐书谦,等.单抗Mab03.2C1C2影响白念珠菌芽管形成及粘附作用的实验研究[J].第三军医大学学报,2005,27(09):901.
[10]江凌晓,郭兆彪,陈泽良,等.单克隆抗体修饰玻璃表面制备蛋白质芯片的初步研究[J].第三军医大学学报,2005,27(08):802.

更新日期/Last Update: 2011-04-13