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.Electronic supplementary materialThe online version of this article (doi:10.1186/s12935-014-0143-3) contains supplementary material, which is available to authorized users.
Highlights
The incidence of head and neck squamous cell carcinoma (HNSCC) varies worldwide but is more prevalent in South Asia [1,2]
The commonly observed presence of neck lymph node metastasis (LNM) in oral squamous cell carcinoma (OSCC) patients signifies the regional spread of the disease
The biopsies from hyperplastic, non-metastatic and metastatic oral tongue squamous cell carcinoma (OTSCC) were incubated in conditioned media to observe their sphere formation patterns in primary cell culture (Table 2)
Summary
The incidence of head and neck squamous cell carcinoma (HNSCC) varies worldwide but is more prevalent in South Asia [1,2]. The commonly observed presence of neck lymph node metastasis (LNM) in oral squamous cell carcinoma (OSCC) patients signifies the regional spread of the disease. The first evidence for the existence of CSCs was reported in blood borne cancer based on expression of CD34 + CD38- markers [5]. This was followed by their identification in solid tumor cancers including breast (CD44 + CD24-) [6,7], brain (CD133+) [8], prostate (CD44+) [9] and pancreatic tumor (CD44 + CD24 + ESA+) [10]
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