Characterization of sphere-forming HCT116 clones by whole RNA sequencing.
10.4174/astr.2016.90.4.183
- Author:
Eunkyung CHUNG
1
;
Inkyung OH
;
Kil Yeon LEE
Author Information
1. R&D Center, L&K Biomed Co. Ltd., Seoul, Korea.
- Publication Type:Original Article
- Keywords:
Neoplastic stem cell;
Colon neoploasms;
RNA sequence analysis;
HCT116 cells
- MeSH:
Cell Separation;
Clinical Coding;
Clone Cells*;
Colonic Neoplasms;
HCT116 Cells;
Humans;
Neoplastic Stem Cells;
Parents;
RNA*;
Sequence Analysis, RNA*;
Transcriptome
- From:Annals of Surgical Treatment and Research
2016;90(4):183-193
- CountryRepublic of Korea
- Language:English
-
Abstract:
PURPOSE: To determine CD133+ cells defined as cancer stem cells (CSCs) in colon cancer, we examined whether CD133+ clones in HCT116 demonstrate known features of CSCs like sphere-forming ability, chemodrug-resistance, and metastatic potential. METHODS: Magnetic cell isolation and cell separation demonstrated that <1% of HCT116 cells expressed CD133, with the remaining cells being CD133- clones. In colon cancer cells, radioresistance is also considered a CSC characteristic. We performed clonogenic assay using 0.4 Gy γ-irradiation. RESULTS: Interestingly, there were no differences between HCT116 parental and HCT116 CD133+ clones when the cells comprised 0.5% of the total cells, and CD133- clone demonstrated radiosensitive changes compared with parental and CD133+ clones. Comparing gene expression profiles between sphere-forming and nonforming culture conditions of HCT116 subclones by whole RNA sequencing failed to obtain specific genes expressed in CD133+ clones. CONCLUSION: Despite no differences of gene expression profiles in monolayer attached culture conditions of each clone, sphere-forming conditions of whole HCT116 subclones, parental, CD133+, and CD133- increased 1,761 coding genes and downregulated 1,384 genes related to CSCs self-renewal and survival. Thus, spheroid cultures of HCT116 cells could be useful to expand colorectal CSCs rather than clonal expansion depending on CD133 expressions.
