Tang Ming-Rui, Guo Jia-Yan, Wang Di, Xu Nan
Department of Plastic Surgery, The First Hospital of China Medical University, Shenyang 110001, People's Republic of China.
Onco Targets Ther. 2018 Jun 12;11:3401-3406. doi: 10.2147/OTT.S157043. eCollection 2018.
Cutaneous melanoma (CM) is a common skin cancer. Surgery is still the primary treatment for CM, as melanoma is resistant to chemotherapy. In the recent years, it has been found that cancer stem-like cells (CSCs) are responsible for this drug resistance. CD24 is a widely used marker to isolate CSCs. In this study, we aimed to analyze the properties of CD24 and CD24 subpopulation of melanoma cells.
We isolated CD24 cells CSCs using magnetic-activated cell sorting system. We extracted total RNA and carried out reverse transcription polymerase chain reaction analysis. We counted the cell colonies using soft agar assay and assessed the cell invasion using cell migration assay. We implanted CD24 or CD24 cells into the flank of non-obese diabetic severe combined immunodeficiency mice, and measured the tumor volumes every 5 days until the end of the experiment. We carried out immunohistochemical analysis to study the tissue sections.
We demonstrated that the CD24 subpopulation has self-renewal properties in vitro and in vivo by using soft agar assay and xenograft tumor model. Furthermore, we confirmed that CD24 expression is accompanied by activation of Notch1 signaling pathway.
This study provides new knowledge on the role of CD24 in the tumorigenic ability of melanoma.
皮肤黑色素瘤(CM)是一种常见的皮肤癌。手术仍然是CM的主要治疗方法,因为黑色素瘤对化疗耐药。近年来,已发现癌症干细胞样细胞(CSCs)是这种耐药性的原因。CD24是一种广泛用于分离CSCs的标志物。在本研究中,我们旨在分析黑色素瘤细胞中CD24及其亚群的特性。
我们使用磁激活细胞分选系统分离CD24细胞CSCs。我们提取总RNA并进行逆转录聚合酶链反应分析。我们使用软琼脂试验计数细胞集落,并使用细胞迁移试验评估细胞侵袭。我们将CD24或CD24细胞植入非肥胖糖尿病严重联合免疫缺陷小鼠的侧腹,并每5天测量一次肿瘤体积,直至实验结束。我们进行免疫组织化学分析以研究组织切片。
通过软琼脂试验和异种移植肿瘤模型,我们证明了CD24亚群在体外和体内具有自我更新特性。此外,我们证实CD24表达伴随着Notch1信号通路的激活。
本研究为CD24在黑色素瘤致瘤能力中的作用提供了新知识。