Department of Otolaryngology-Head and Neck Surgery, Eye, Ear, Nose and Throat Hospital, Fudan University, Shanghai, People's Republic of China.
PLoS One. 2013 Jun 11;8(6):e65750. doi: 10.1371/journal.pone.0065750. Print 2013.
Cancer stem-like side population (SP) cells have been identified in many solid tumors; however, most of these investigations are performed using established cancer cell lines. Cancer cells in tumor tissue containing fibroblasts and many other types of cells are much more complex than any cancer cell line. Although SP cells were identified in the laryngeal squamous cell carcinoma (LSCC) cell line Hep-2 in our pilot study, it is unknown whether the LSCC tissue contains SP cells. In this study, LSCC cells (LSCCs) were primary cultured and purified from a surgically resected LSCC specimen derived from a well-differentiated epiglottic neoplasm of a Chinese male. This was followed by the verification of epithelium-specific characteristics, such as ultrastructure and biomarkers. A distinct SP subpopulation (4.45±1.07%) was isolated by Hoechst 33342 efflux analysis from cultured LSCCs by using a flow cytometer. Cancer stem cell (CSC)-associated assays, including expression of self-renewal and CSC marker genes, proliferation, differentiation, spheroid formation, chemotherapy resistance, and tumorigenicity were then conducted between SP and non-SP (NSP) LSCCs. In vitro and in vivo assays revealed that SP cells manifested preferential expression of self-renewal and CSC marker genes, higher capacity for proliferation, differentiation, and spheroid formation; enhanced resistance to chemotherapy; and greater xenograft tumorigenicity in immunodeficient mice compared with NSP cells. These findings suggest that the primary cultured and purified LSCCs contain cancer stem-like SP cells, which may serve as a valuable model for CSC research in LSCC.
癌症干细胞样侧群 (SP) 细胞已在许多实体瘤中被鉴定出来；然而，大多数这些研究是使用已建立的癌细胞系进行的。肿瘤组织中的癌细胞含有成纤维细胞和许多其他类型的细胞，比任何癌细胞系都要复杂得多。尽管在我们的初步研究中在喉鳞状细胞癌 (LSCC) 细胞系 Hep-2 中鉴定出了 SP 细胞，但尚不清楚 LSCC 组织中是否含有 SP 细胞。在这项研究中，LSCC 细胞（LSCCs）是从一位中国男性分化良好的会厌肿瘤手术切除的 LSCC 标本中进行原代培养和纯化的。随后验证了上皮细胞的特异性特征，如超微结构和生物标志物。通过使用流式细胞仪，从培养的 LSCC 中通过 Hoechst 33342 外排分析分离出明显的 SP 亚群（4.45±1.07%）。然后，在 SP 和非 SP（NSP）LSCC 之间进行了与癌症干细胞 (CSC) 相关的测定，包括自我更新和 CSC 标记基因的表达、增殖、分化、球体形成、化疗耐药性和肿瘤发生。体外和体内测定表明，SP 细胞表现出自我更新和 CSC 标记基因的优先表达、更高的增殖、分化和球体形成能力、增强的化疗耐药性以及在免疫缺陷小鼠中的异种移植物肿瘤发生能力。这些发现表明，原代培养和纯化的 LSCC 含有癌症干细胞样 SP 细胞，可作为 LSCC CSC 研究的有价值模型。
