CD44high/ALDH1high head and neck squamous cell carcinoma cells exhibit mesenchymal characteristics and GSK3-dependent cancer stem cell properties

BackgroundCD44 and aldehyde dehydrogenase 1 (ALDH1) have been shown to be useful markers for identification of cancer stem cells (CSCs). We previously reported that glycogen synthase kinase 3 (GSK3) is involved in regulation of the self-renewal ability of head and neck squamous cell carcinoma (HNSCC) CSCs. The purpose of the present study was to clarify the role of GSK3 in CD44(high)/ALDH1(high) HNSCC cells. MethodsCells with greater expression of CD44 and higher ALDH1 enzymatic activity were FACS sorted from the OM-1 HNSCC cell line. The self-renewal ability of CD44(high)/ALDH1(high) cells was then examined using a tumor sphere formation assay. mRNA expressions of the stem cell markers Sox2, Oct4, and Nanog, as well as GSK3 were evaluated by real-time RT-PCR. ResultsCD44(high)/ALDH1(high) cells exhibited higher tumor sphere forming ability and increased expression of stem cell markers as compared with CD44(high)/ALDH1(low) cells. Interestingly, spindle-shaped cells positive for vimentin were found in the CD44(high)/ALDH1(high) but not the CD44(high)/ALDH1(low) cell population. In addition, the ALDH1 activity and sphere forming ability of CD44(high)/ALDH1(high) cells was significantly inhibited by GSK3 knockdown. On the other hand, CD44(high)/ALDH1(low) cells exhibited high epidermal growth factor receptor (EGFR) expression and increased cell growth. ConclusionsOur results show that GSK3 plays a major role in maintenance of stemness of CD44(high)/ALDH1(high) HNSCC cells. Additionally, they indicate a close relationship between CSC and mesenchymal characteristics in HNSCC.