Molecular Pathology Laboratory, Department of Cytology and Gynecological Pathology, Postgraduate Institute of Medical Education and Research, Chandigarh, PIN-160012, India.
Department of Obstetrics and Gynecology, Postgraduate Institute of Medical Education and Research, Chandigarh, India.
BMC Cancer. 2018 Apr 2;18(1):357. doi: 10.1186/s12885-018-4237-5.
Cervical cancer is a major cause of cancer-related mortality in women in the developing world. Cancer Stem cells (CSC) have been implicated in treatment resistance and metastases development; hence understanding their significance is important.
Primary culture from tissue biopsies of invasive cervical cancer and serial passaging was performed for establishing cell lines. Variable Number Tandem Repeat (VNTR) assay was performed for comparison of cell lines with their parental tissue. Tumorsphere and Aldefluor assays enabled isolation of cancer stem cells (CSC); immunofluorescence and flow cytometry were performed for their surface phenotypic expression in cell lines and in 28 tissue samples. Quantitative real-time PCR for stemness and epithelial-mesenchymal transition (EMT) markers, MTT cytotoxicity assay, cell cycle analysis and cell kinetic studies were performed.
Four low-passage novel cell lines designated RSBS-9, - 14 and - 23 from squamous cell carcinoma and RSBS-43 from adenocarcinoma of the uterine cervix were established. All were HPV16+. VNTR assay confirmed their uniqueness and derivation from respective parental tissue. CSC isolated from these cell lines showed CD133 phenotype. In tissue samples of untreated invasive cervical cancer, CD133 CSCs ranged from 1.3-23% of the total population which increased 2.8-fold in radiation-resistant cases. Comparison of CD133 with CD133 bulk population cells revealed increased tumorsphere formation and upregulation of stemness and epithelial-mesenchymal transition (EMT) markers with no significant difference in cisplatin sensitivity.
Low-passage cell lines developed would serve as models for studying tumor biology. Cancer Stem Cells in cervical cancer display CD133 phenotype and are increased in relapsed cases and hence should be targeted for achieving remission.
宫颈癌是发展中国家女性癌症相关死亡的主要原因。癌症干细胞（CSC）被认为与治疗耐药性和转移发展有关；因此，了解其意义很重要。
对浸润性宫颈癌组织活检进行原代培养和连续传代，建立细胞系。可变串联重复（VNTR）分析用于比较细胞系与其亲本组织。肿瘤球和 Aldefluor 分析可分离癌症干细胞（CSC）；免疫荧光和流式细胞术用于细胞系和 28 个组织样本中 CSC 的表面表型表达。进行了干细胞和上皮-间充质转化（EMT）标志物的实时定量 PCR、MTT 细胞毒性测定、细胞周期分析和细胞动力学研究。
从鳞状细胞癌中建立了四个低传代新型细胞系，命名为 RSBS-9、-14 和-23，从子宫颈腺癌中建立了一个细胞系 RSBS-43。它们均为 HPV16+。VNTR 分析证实了它们的独特性及其源自各自的亲本组织。从这些细胞系中分离的 CSC 表现出 CD133 表型。在未经治疗的浸润性宫颈癌组织样本中，CD133 CSC 占总细胞群的 1.3-23%，在放疗耐药病例中增加了 2.8 倍。与 CD133 bulk 群体细胞比较显示，CD133 肿瘤球形成增加，干细胞和上皮-间充质转化（EMT）标志物上调，顺铂敏感性无显著差异。
建立的低传代细胞系将作为研究肿瘤生物学的模型。宫颈癌中的癌症干细胞表现出 CD133 表型，在复发病例中增加，因此应作为实现缓解的靶点。
