正常与乳腺肿瘤细胞表面黏附分子CD44活化状态差异分析

doi:10.3969/j.issn.1673-8640.2013.05.020

检验医学 ›› 2013, Vol. 28 ›› Issue (5): 425-429.DOI: 10.3969/j.issn.1673-8640.2013.05.020

正常与乳腺肿瘤细胞表面黏附分子CD44活化状态差异分析

侯利丹,刘鹥雯,何怡青,杨翠霞,杜艳,　　　　　　　　　高锋

上海交通大学附属第六人民医院中心实验室，上海 200233

收稿日期:2013-01-22 修回日期:2013-02-01 出版日期:2013-05-15 发布日期:2013-05-14
通讯作者: 高锋，联系电话：021-64369181。
作者简介:侯利丹，女，1988年生，硕士，主要从事肿瘤靶向治疗研究。
基金资助:
国家自然科学基金（No.81071814、81172027、81272479）；上海市科委重点项目（No.10411950500）；上海市优秀学术带头人计划（No.11XD1404000）

Analysis on the differences of surface adhesion molecule CD44 activation states in normal cells and breast tumor cells

Department of Central Laboratory，Shanghai Sixth People′s Hospital，Shanghai Jiaotong University，Shanghai 200233，China

Received:2013-01-22 Revised:2013-02-01 Online:2013-05-15 Published:2013-05-14

摘要/Abstract

摘要： 摘要：目的　检测黏附分子CD44在正常和乳腺肿瘤细胞表面的表达，并分析其活化状态差异，为以CD44为靶点的肿瘤靶向治疗提供新的理论依据。方法　采用流式细胞术检测20名正常人外周血单个核细胞（PBMCs）、正常小鼠胚细胞NIH3T3及人乳腺肿瘤细胞Hs578T、BT-549表面CD44表达水平；使用荧光标记透明质酸(FL-HA)，通过流式细胞术分析CD44的活化状态；再运用细胞免疫荧光的方法观察HA与不同活化状态CD44的靶向结合情况。结果　正常人PBMCs、NIH3T3与乳腺肿瘤细胞Hs578T、BT-549表面均高表达CD44，阳性表达率＞95%。PBMCs、NIH3T3的CD44与FL-HA结合活性相对较低，分别为（3.61±2.65）% 和（14.33±1.35）%，而乳腺肿瘤细胞表面CD44与FL-HA结合活性高（＞90%）。细胞免疫荧光显示NIH3T3细胞表面基本无荧光，HA不靶向结合CD44，而Hs578T和BT-549细胞表面呈现强荧光，HA靶向结合CD44。结论　正常细胞PBMCs、NIH3T3表面虽然高表达CD44，但与其天然配体HA结合活性较低，而乳腺肿瘤细胞表面高表达CD44且与其配体HA结合活性很高。提示CD44分子在正常和肿瘤细胞表面存在相对静止与活化2种状态，前者与HA结合活性较低，后者与HA结合活性高。提示以HA为靶向分子、CD44为靶点的抗肿瘤药物，能够选择性杀伤高表达CD44的肿瘤细胞而不影响正常细胞。

关键词: CD44, 活化状态, 乳腺肿瘤, 外周血单个核细胞

Abstract: Objective　To detect the expressions of surface adhesion molecule CD44 in normal cells and breast tumor cells， and analyze the differences of CD44 activation states in order to provide a new theoretical reference for CD44 targeted cancer therapy. Methods　The expressions of CD44 in normal peripheral blood mononuclear cells（PBMCs），normal mouse fibroblast NIH3T3 cells and human breast tumor cell Hs578T and BT-549 were determined by flow cytometry. The activation states of CD44 were analyzed by flow cytometry， using fluorescence-labeled hyaluronic acid（FL-HA）， and then the targeted binding ability of HA-CD44 was observed by cell immunofluorescence. Results　CD44 was abundantly expressed in normal PBMCs， NIH3T3 and breast tumor cell Hs578T and BT-549 (the positive expression rate was ＞95%). The binding ability of the breast tumor cell surface CD44 with FL-HA (＞90%) was much stronger than those of PBMCs[(3.61±2.65)%] and NIH3T3[(14.33±1.35)%]. NIH3T3 cell showed almost no fluorescence(HA targeted binding without CD44)， while Hs578T and BT-549 showed strong fluorescence (HA targeted binding with CD44)according to cell immunofluorescence. Conclusions　Normal cell PBMCs and NIH3T3 have high expressions of CD44 and low binding ability with HA， while breast tumor cell Hs578T and BT-549 have high expressions of CD44 and high binding ability with other HA. CD44 in normal cells and breast tumor cells shows 2 different activation states：inactive and active. The anti-tumor drugs with HA as targeted mononuclear and CD44 as target can select high-expression CD44 killer tumor cells and have no influence on normal cells.

Key words: CD44, Activation state, Breast tumor, Peripheral blood mononuclear cell

侯利丹，刘鹥雯，何怡青，杨翠霞，杜艳，高锋. 正常与乳腺肿瘤细胞表面黏附分子CD44活化状态差异分析[J]. 检验医学, 2013, 28(5): 425-429.

HOU Lidan，LIU Yiwen，HE Yiqing，YANG Cuixia，DU Yan，GAO Feng. Analysis on the differences of surface adhesion molecule CD44 activation states in normal cells and breast tumor cells[J]. , 2013, 28(5): 425-429.

参考文献

[1]Orian-Rousseau V. CD44， a therapeutic target for metastasising tumours[J]. Eur J Cancer， 2010，46(7):1271-1277.
[2] Toole BP，Slomiany MG. Hyaluronan，CD44 and Emmprin: partners in cancer cell chemoresistance[J]. Drug Resist Updat，2008 ，11(3):110-121.
[3]Coussens LM， Werb Z. Inflammatory cells and cancer: think different[J]. J Exp Med ， 2001，193(6): F23-F26.
[4] Ponta H， Sherman L， Herrlich PA. CD44: from adhesion molecules to signalling regulators[J]. Nat Rev Mol Cell Biol，2003，4(1):33-45.
[5]Zawadzki V， Perschl A， Rsel M， et al.Blockade of metastasis formation by CD44-receptor globulin[J]. Int J Cancer， 1998，75(6): 919-924.
[6]Auzenne E， Ghosh SC， Khodadadian M， et al. Hyaluronic acid-paclitaxel: antitumor efficacy against CD44 ( ) human ovarian carcinoma xenografts[J]. Neoplasia， 2007， 9(6): 479-486.
[7] Journo-Gershfeld G， Kapp D， Shamay Y， et al. Hyaluronan oligomers-HPMA copolymer conjugates for targeting paclitaxel to CD44-overexpressing ovarian carcinoma[J]. Pharm Res，2012，29(4):1121-1133.
[8]Cho HJ， Yoon IS， Yoon HY，et al. Polyethylene glycol-conjugated hyaluronic acid-ceramide self-assembled nanoparticles for targeted delivery of doxorubicin[J]. Biomaterials，2012 ，33(4):1190-1200.
[9]Rodgers AK， Nair A， Binkley PA，et al. Inhibition of CD44 N- and O-linked glycosylation decreases endometrial cell lines attachment to peritoneal mesothelial cells[J]Fertil Steril， 2011，95(2):823-825.
[10] Tzircotis G， Thorne RF， Isacke CM. Chemotaxis towards hyaluronan is dependent on CD44 expression and modulated by cell type variation in CD44-hyaluronan binding[J]. J Cell Sci， 2005，118(Pt21): 5119-5128.
[11]Lesley J， Gál I， Mahoney DJ， et al. TSG-6 modulates the interaction between hyaluronan and cell surface CD44[J]. J Biol Chem， 2004， 279(24): 25745-25754.
[12]Richter U， Wicklein D， Geleff S， et al. The inter-action between CD44 on tumor cells and hyaluronan under physiologic flow conditions: implications for metastasis formation[J]. Histochem Cell Biol，2012，137(5):687-695.
[13]Hanagiri T， Shinohara S， Takenaka M， et al. Effects of hyaluronic acid and CD44 interaction on the proliferation and invasiveness of malignant pleural mesothelioma[J]. Tumour Biol，2012，33(6):2135-2341.

[1]	杨艳敏, 彭梦乐, 连文萍. 直肠癌患者MDSC百分比和RORα mRNA、IL-17E mRNA、FOXP3 mRNA表达的临床意义[J]. 检验医学, 2023, 38(1): 23-27.
[2]	刘蓉, 王德刚. 卵巢癌组织中SIRT-1、CD44的表达及临床意义[J]. 检验医学, 2022, 37(7): 632-6358.
[3]	何亚亚, 陈闪闪, 李永胜, 刘娟. AS患者sLAIR-1和LAIR-1 mRNA的表达及其与疾病活动度的关系[J]. 检验医学, 2022, 37(10): 915-920.
[4]	苏曦, 周琰, 沈敏娜, 李懿皞, 王蓓丽, 潘柏申, 郭玮. 不同样本类型对EBV DNA检测结果的影响[J]. 检验医学, 2020, 35(8): 803-805.
[5]	王春玮, 蔡星垚, 张世宽, 郭景景. 不同样本中EBV DNA检出率的差异[J]. 检验医学, 2019, 34(8): 763-764.
[6]	张翠. 微小RNA-132和高迁移率族蛋白1在脓毒症患儿外周血单个核细胞中的表达及临床意义[J]. 检验医学, 2018, 33(9): 815-818.
[7]	丁冰玉, 林萍, 李擎天. 植物血凝素刺激外周血单个核细胞Th1/Th2细胞因子分泌对阿尔茨海默病的诊断价值[J]. 检验医学, 2018, 33(4): 290-294.
[8]	姜小华, 卫宇, 孙建文, 任孔华, 崔北年, 汪伟, 程怡. 原发性乳腺癌AS160分子磷酸化水平与肿瘤细胞增殖活性指标的相关性分析[J]. 检验医学, 2016, 31(3): 220-223.
[9]	肖敏敏，王毅，邵慧，张艳，李萍. 皖南地区HIV感染者中HCV感染状况的调查[J]. 检验医学, 2014, 29(7): 705-707.
[10]	黄红莉，周刚，牛立志，石红霞，孔小峰，王峰，徐克成. 胰腺癌患者冷冻消融治疗前后血清CD44v6和整合素β1的变化及在预后评估中的意义[J]. 检验医学, 2013, 28(12): 1083-1087.
[11]	刘阳，胡梅，喻红波，黎健. 肝癌患者外周血单个核细胞HLA-G表达研究[J]. 检验医学, 2013, 28(12): 1088-1091.
[12]	钱　雷;汪晓莺;吕丽君;陈德训 . 类风湿性关节炎患者早期外周血单个核细胞Toll样受体2、4表达及意义[J]. 检验医学, 2012, 27(8): 659-662.
[13]	王翠玲;张宜俊;李茹冰;王增松;夏书奇;段祥;谢飞群;曾振飞;揭燕;傅泳航. HBsAg联合rhIL-12对慢性乙型肝炎患者PBMC诱生IFN-γ的作用[J]. 检验医学, 2009, 24(09): 671-675.
[14]	王颖智;杨翠霞;刘鷖雯;何怡青;杨思;崔练;高锋. 实时荧光定量PCR检测CD44和RHAMM在胃肠肿瘤中的表达研究[J]. 检验医学, 2008, 23(04): 338-343.
[15]	钱(王争);邓安梅;陈孙孝;王燕;周晔;陈燕;谷明莉;仲人前. 人颗粒溶素的克隆、原核表达及其抗体制备[J]. 检验医学, 2007, 22(05): 571-574.

正常与乳腺肿瘤细胞表面黏附分子CD44活化状态差异分析

Analysis on the differences of surface adhesion molecule CD44 activation states in normal cells and breast tumor cells

RichHTML

PDF (PC)

可视化

被引次数

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