%0 Journal Article
%T Decreased expression of cell adhesion genes in cancer stem
%A Amrendra Mishra
%A Ashok Gopinath
%A Harshini Sriram
%A Pinal Chandarana
%A Raman Govindarajan
%A Rekha V Kumar
%A S. Ramaswamy
%A Subhashini Sadasivam
%A Vivek Tanavde
%J Tumor Biology
%@ 1423-0380
%D 2018
%R 10.1177/1010428318780859
%X The goal of this study was to isolate cancer stem-like cells marked by high expression of CD44, a putative cancer stem cell marker, from primary oral squamous cell carcinomas and identify distinctive gene expression patterns in these cells. From 1 October 2013 to 4 September 2015, 76 stage III每IV primary oral squamous cell carcinoma of the gingivobuccal sulcus were resected. In all, 13 tumours were analysed by immunohistochemistry to visualise CD44-expressing cells. Expression of CD44 within The Cancer Genome Atlas每Head and Neck Squamous Cell Carcinoma RNA-sequencing data was also assessed. Seventy resected tumours were dissociated into single cells and stained with antibodies to CD44 as well as CD45 and CD31 (together referred as Lineage/Lin). From 45 of these, CD44+Lin每 and CD44每Lin每 subpopulations were successfully isolated using fluorescence-activated cell sorting, and good-quality RNA was obtained from 14 such sorted pairs. Libraries from five pairs were sequenced and the results analysed using bioinformatics tools. Reverse transcription quantitative polymerase chain reaction was performed to experimentally validate the differential expression of selected candidate genes identified from the transcriptome sequencing in the same 5 and an additional 9 tumours. CD44 was expressed on the surface of poorly differentiated tumour cells, and within the The Cancer Genome Atlas每Head and Neck Squamous Cell Carcinoma samples, its messenger RNA levels were higher in tumours compared to normal. Transcriptomics revealed that 102 genes were upregulated and 85 genes were downregulated in CD44+Lin每 compared to CD44每Lin每 cells in at least 3 of the 5 tumours sequenced. The upregulated genes included those involved in immune regulation, while the downregulated genes were enriched for genes involved in cell adhesion. Decreased expression of PCDH18, MGP, SPARCL1 and KRTDAP was confirmed by reverse transcription quantitative polymerase chain reaction. Lower expression of the cell每cell adhesion molecule PCDH18 correlated with poorer overall survival in the The Cancer Genome Atlas每Head and Neck Squamous Cell Carcinoma data highlighting it as a potential negative prognostic factor in this cancer
%K Oral squamous cell carcinoma
%K gingivobuccal sulcus
%K RNA-sequencing
%K cancer stem cells
%K CD44
%K PCDH18
%U https://journals.sagepub.com/doi/full/10.1177/1010428318780859