Radiation Biology & Health Sciences Division, Bhabha Atomic Research Centre, Mumbai 400 085, India.
Radiation Biology & Health Sciences Division, Bhabha Atomic Research Centre, Mumbai 400 085, India; Homi Bhabha National Institute, Anushakti Nagar, Mumbai 400 094, India.
Tissue Cell. 2019 Oct;60:21-24. doi: 10.1016/j.tice.2019.07.003. Epub 2019 Jul 16.
Cancer stem cells (CSCs) have been implicated in growth, metastasis, recurrence and chemo-/radio-resistance in several cancer types. Despite a plenty of literature about different in vitro techniques to enrich/isolate CSCs, their comparative characterization for stemness is not well established. In the present study, cells obtained following three in vitro assays [clonogenic assay, tumorsphere assay (TSA) and single cell assay (SCA)] were compared for their cancer stem-like cell characteristics in human lung adenocarcinoma (A549) cells. Expression of the pluripotent (OCT4, NANOG) and lung cancer stem cell marker (CD166) genes were studied in these cells. Results showed that in comparison to cells obtained from routine culture (CC), the cells obtained from TSA showed significantly higher expression of OCT-4 and NANOG. These results were further validated with quantification of cell surface cancer stem cell markers i.e. CD44/CD24 in the cells obtained from different methods, which were higher in TSA and SCA. Additionally, functional characterization of cancer stem-like cells (CSLCs) using ALDH assay showed the highest % of ALDH cells in TSA. These results were in agreement with higher resistance of these cells against 5-Fluorouracil suggesting higher fraction of CSLCs in TSA than the other assays. These results showed that TSA provides a better method to enrich CSLCs in A549 lung adenocarcinoma cells.
癌症干细胞 (CSCs) 被认为与多种癌症的生长、转移、复发和化疗/放疗耐药有关。尽管有大量关于不同体外技术富集/分离 CSCs 的文献，但它们在干性方面的比较特征尚未得到很好的建立。在本研究中，比较了三种体外检测方法（克隆形成检测、肿瘤球检测（TSA）和单细胞检测（SCA））获得的细胞在人肺腺癌细胞（A549）中的癌症干细胞样细胞特征。研究了这些细胞中多能（OCT4、NANOG）和肺癌干细胞标志物（CD166）基因的表达。结果表明，与常规培养（CC）获得的细胞相比，TSA 获得的细胞中 OCT-4 和 NANOG 的表达显著升高。这些结果通过定量分析不同方法获得的细胞表面癌症干细胞标志物即 CD44/CD24 得到进一步验证，TSA 和 SCA 中的细胞更高。此外，使用 ALDH 检测对癌症干细胞样细胞（CSLCs）进行功能表征显示，TSA 中具有最高比例的 ALDH 细胞。这些结果与这些细胞对 5-氟尿嘧啶的更高抗性一致，表明 TSA 中的 CSLCs 比例高于其他检测方法。这些结果表明，TSA 为在 A549 肺腺癌细胞中富集 CSCs 提供了一种更好的方法。
