Evaluation of stem-like side population cells in a recurrent nasopharyngeal carcinoma cell line

Cancer Cell International

Alan Soo Beng Khoo³  Munirah Ahmad³  Wen Cai Zhang⁴  Ching Ching Ng¹  Suat Cheng Peh⁵  Juliana Jamal²  Lu Ping Tan³  Susan Ling Ling Hoe¹ 

[1] Institute of Biological Sciences, Faculty of Science, University of Malaya, Kuala Lumpur, 50603, Malaysia;Current address: Faculty of Medicine, University of Malaya, Kuala Lumpur, 50603, Malaysia;Molecular Pathology Unit, Cancer Research Centre, Institute for Medical Research, Jalan Pahang, Kuala Lumpur, 50588, Malaysia;Genome Institute of Singapore, 60 Biopolis Street, #02-01, Genome 138672, Singapore;Faculty of Medical Sciences, UCSI University, 1 Jalan Menara Gading, UCSI Heights, Cheras, 56000, Malaysia

关键词: Cancer stem cells;    Stem-like;    Slow-cycling;    Nasopharyngeal carcinoma;    Side population;   

Others  :  1121650
DOI  :  10.1186/s12935-014-0101-0

 received in 2014-05-19, accepted in 2014-09-22,  发布年份 2014

【 摘 要 】

Background

Side population (SP) assay identifies cells with dye/drug extrusion ability, a characteristic of stem cells. Here, we determined if SP cells exist in a verified cell line originating from recurrent nasopharyngeal carcinoma (NPC) and a xenograft established from recurrent metastatic NPC. These cells were evaluated for stem-like properties via functional assays as well as for tumourigenicity.

Methods

We used Hoechst 33342 to identify the SP from non-SP (NSP) cells in HK1 NPC cell line and xeno-284 NPC xenograft. The cells were assayed for in vitro characteristics of cancer stem cells (CSC), gene expression and tumourigenicity ability. Student’s t test was used to test for significance.

Results

Five to ten percent and less than 0.5% of HK1 and xeno-284 NPC cells, respectively, were SP cells. Fumitremorgin C (FTC), as opposed to verapamil, was effective in causing the cells to retain Hoechst 33342 dye. HK1 SP cells formed more holoclones, had more aldehyde dehydrogenase (ALDH) activity, divided asymmetrically and contained slow-proliferating cells. ABCG2, SOX2, TERT, MYC, Hedgehog, Notch, TGFβ and Wnt signalling pathway genes were significantly upregulated in the SP cells. However, despite these differences in vitro, both HK1 SP and NSP cells had an overall similar tumourigenic potential in vivo.

Conclusions

HK1 SP cells were ABCG2-specific as confirmed by FTC inhibition and gene expression data. Despite data from in vitro and gene expression experiments suggesting stem-like features, there was no significant difference in tumourigenic potential between SP and NSP cells. We conclude that SP assay alone is not sufficient to identify CSCs in HK1 cells. Our work also suggests the presence of a stem-cell like population among NPC cells which do not display increased tumourigenicity.

【 授权许可】

   
2014 Hoe et al.; licensee BioMed Central Ltd.

【 图 表 】

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