Various malignant cancers have been found to contain a sub-population of stem cell-like tumour cells, or cancer stem cells (CSCs), however, culture methods for CSCs and the size of the fraction of CSCs in C6, which is a commonly used glioma cell line, remain controversial. In this study, we demonstrated that the C6 cell line contains a fraction of tumour cells that can form tumour spheres in a simplified serum-free neural stem cell medium and express CD133 and nestin, which are widely-used markers for brain CSCs. Immunohistochemistry and immunofluorescence confirmed the existence of CSCs both in the C6 cell line and C6 xenografts. Flow cytometry demonstrated that 4.02% of cells in the C6 cell line and 4.21% in the C6 xenografts presented as CSCs. These results confirm the fraction of CSCs in the C6 cell line and provide a simple and effective method for isolation of CSCs to study the initiation and progression of human glioma and, possibly, other malignant tumours.

This study compared clinical features and protein expression profiles in differentiated thyroid tumours to identify protein markers with the potential for indicating malignancy status. Tissue microarrays were constructed using 119 thyroid tumour samples (45 papillary carcinomas, 26 follicular carcinomas, 48 adenomas). Generally, there was overexpression of proliferating cell nuclear antigen (PCNA), p53, matrix metalloproteinase (MMP)-7, Hector Battifora mesothelial-1 (HBME-1), MMP-2, pituitary tumour-transforming gene (PTTG) and human telomerase reverse transcriptase (hTERT) in malignant thyroid carcinomas, and overexpression of fragile histidine triad (FHIT), p16 and Ecadherin in thyroid adenomas. Multiple factor binary logistic regression analysis indicated that MMP-2, HBME-1, p16 and FHIT were independently related to differentiated thyroid tumours. Receiveroperating characteristics for these four factors showed HBME-1 as best for diagnostic accuracy. Sensitivity and specificity were enhanced using an HBME-1 and p16 cluster. HBME-1 expression was not significantly different for papillary and follicular carcinomas, whereas p16 expression was significantly specific.