title: Patterns of cancer cell sphere formation in primary cultures of human oral tongue squamous cell carcinoma and neck nodes.
abstract: Recently a sub-population of cells with stem cell characteristics, reported to be associated with initiation, growth, spread and recurrence, has been identified in several solid tumors including oral tongue squamous cell carcinoma (OTSCC). The aim of our pilot study was to isolate CD44+ cancer stem cells from primary cultures of OTSCC and neck node Level I (node-I) biopsies, grow cell spheres and observe their characteristics in primary cultures. Parallel cultures of hyperplastic lesions of tongue (non-cancer) were set up as a control. Immunohistochemistry was used to detect CD44/CD24 expression and magnetic activated cell sorting to isolate CD44+ cell populations followed by primary cell culturing. Both OTSCC and node-I biopsies produced floating spheres in suspension, however those grown in hyperplastic and node-I primary cultures did not exhibit self-renewal properties. Lymph node metastatic OTSCC, express higher CD44/CD24 levels, produce cancer cell spheres in larger number and rapidly (24 hours) compared to node negative OTSCC (1 week) and non-cancer specimens (3 weeks). In addition, metastatic OTSCC have the capacity for proliferation for up to three generations in primary culture. This in vitro system will be used to study cancer stem cell behavior, therapeutic drug screening and optimization of radiation dose for elimination of resistant cancer cells.
authors: Basic Sciences Research Department, Shaukat Khanum Memorial Cancer Hospital and Research Center, 7-A Block R-3, Johar Town, Lahore, 54000 Pakistan.
authors: Saira
authors: S
authors: Saleem
authors: Department of Radiation Oncology, Shaukat Khanum Memorial Cancer Hospital and Research Center, 7-A Block R-3, Johar Town, Lahore, 54000 Pakistan.
authors: Arif
authors: A
authors: Jamshed
authors: Clinical Research Office, Shaukat Khanum Memorial Cancer Hospital and Research Center, 7-A Block R-3, Johar Town, Lahore, 54000 Pakistan.
authors: Saima
authors: S
authors: Faisal
authors: Department of Surgical Oncology, Shaukat Khanum Memorial Cancer Hospital and Research Center, 7-A Block R-3, Johar Town, Lahore, 54000 Pakistan.
authors: Raza
authors: R
authors: Hussain
authors: Department of Pathology, Shaukat Khanum Memorial Cancer Hospital and Research Center, 7-A Block R-3, Johar Town, Lahore, 54000 Pakistan.
authors: Muhammad
authors: M
authors: Tahseen
authors: Department of Pathology, Shaukat Khanum Memorial Cancer Hospital and Research Center, 7-A Block R-3, Johar Town, Lahore, 54000 Pakistan.
authors: Asif
authors: A
authors: Loya
authors: Institute of Cancer Therapeutics, University of Bradford, Tumbling hill street, Bradford, BD7 1BD United Kingdom.
authors: Chris
authors: C
authors: Sutton
keywords: Cancer cell sphere; Cancer stem cells; Lymph node metastasis; Oral tongue squamous cell carcinoma; Primary culture; in vitro assay