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利用细胞表面标志物CD133和CXCR4分离原发性子宫内膜癌中的干细胞样癌细胞

Isolation of Stem-Like Cancer Cells in Primary Endometrial Cancer Using Cell Surface Markers CD133 and CXCR4.

作者信息

Sun Yi, Yoshida Toshiko, Okabe Motonori, Zhou Kaixuan, Wang Fang, Soko Chika, Saito Sigeru, Nikaido Toshio

机构信息

Department of Regenerative Medicine, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, Japan; Intensive Care Unit, The Affiliated Hospital of Inner Mongolia medical university, China.

Department of Regenerative Medicine, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, Japan.

出版信息

Transl Oncol. 2017 Dec;10(6):976-987. doi: 10.1016/j.tranon.2017.07.007. Epub 2017 Nov 2.

Abstract

Endometrial cancer (EC) is the most common familiar gynecologic malignant tumor identified in the female reproductive system and has been increasing yearly. In this study, we will identify the surface markers and stem cell markers related with cancer stem cells (CSCs) of EC. Tissue samples were obtained from endometrial cancer patients during surgical procedures. Single cells were isolated from the tissues for culturing, transfection into nude mice, and histopathology analysis. RT-PCR demonstrated that the cultured cells strongly expressed stemness-related genes, such as c-Myc, Sox-2, Nanog, Oct 4A, ABCG2, BMI-1, CK-18, Nestin and β-actin. The expression of surface markers CD24, CD133, CD47, CD29, CD44, CXCR4, SSEA3 and SSEA4, CD24, and CD133 and chemokine markers such as CXCR4 were measured by flow cytometry. Then the double percentage of CD133+CXCR4+ cells constituted 7.2% and 9.3% in EC cells originated from two different patients, respectively. The CD133+CXCR4+ primary endometrial cancer cells grew faster, exhibited high expression of mRNA of stemness-related genes, produced more spheres, and had higher clonogenic ability than other subpopulations. They are also more resistant to anti-cancer drugs than other subpopulations. These findings indicate that CD133+CXCR4+ cells may possess some characteristics of CSCs in primary endometrial cancer. These cell surface markers may be useful for the development of drugs against CSC molecular targets or as a predictive marker for poor prognosis in primary endometrial cancer.

摘要

子宫内膜癌(EC)是女性生殖系统中最常见的妇科恶性肿瘤,且其发病率逐年上升。在本研究中,我们将鉴定与子宫内膜癌癌症干细胞(CSCs)相关的表面标志物和干细胞标志物。在手术过程中从子宫内膜癌患者获取组织样本。从组织中分离出单细胞用于培养、接种到裸鼠体内以及进行组织病理学分析。逆转录聚合酶链反应(RT-PCR)表明,培养的细胞强烈表达与干性相关的基因,如c-Myc、Sox-2、Nanog、Oct 4A、ABCG2、BMI-1、CK-18、巢蛋白和β-肌动蛋白。通过流式细胞术检测表面标志物CD24、CD133、CD47、CD29、CD44、CXCR4、阶段特异性胚胎抗原3(SSEA3)和阶段特异性胚胎抗原4(SSEA4)、CD24以及CD133的表达,以及趋化因子标志物如CXCR4的表达。然后,在源自两名不同患者的子宫内膜癌细胞中,CD133+CXCR4+细胞的双阳性率分别为7.2%和9.3%。与其他亚群相比,CD133+CXCR4+原发性子宫内膜癌细胞生长更快,干性相关基因的mRNA表达高,产生更多球体,且具有更高的克隆形成能力。它们对抗癌药物的抗性也比其他亚群更强。这些发现表明,CD133+CXCR4+细胞可能具有原发性子宫内膜癌癌症干细胞的某些特征。这些细胞表面标志物可能有助于开发针对癌症干细胞分子靶点的药物,或作为原发性子宫内膜癌预后不良的预测标志物。

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