Comparison of nucleostemin gene expression in CD133+ and CD133- cell population in colon cancer cell line HT29.
BACKGROUND
Nucleostemin has been shown to be essential for proliferation and survival of colon cancer cells. In this study, we evaluate and comparing nucleostemin expression in CD133+ and CD133- colon cancer cell line HT29.


METHODS
After preparation and culturing of HT29 cell line, isolation was performed using magnetic cell separation system by CD133 MicroBeads and phycoerythrin conjugated to monoclonal anti-human CD133 monoclonal antibody and analyzed with flow cytometry. For quantitative expression of nucleostemin in HT29, CD133+ and CD133- cells used specific nucleostemin primer and glyceraldehyde 3-phosphate dehydrogenase primer as endogenous control.


RESULTS
The results showed the percentage of CD133+ cells in HT29 colon cancer cell line ranged from 36.5% to 41.5%, whereas the percentage of CD133- cell ranged from 58.5% to 63.5%. The expression rate of nucleostemin in HT29, CD133+ and CD133- cells were 1.44 ± 0.78, 1.60 ± 0.70 and 1.00 ± 0.18 (respectively). The comparison of expression rate represents no significant difference in nucleostemin expression in CD133+, CD133- and HT29 colon cancer cells.


CONCLUSIONS
It is concluded that nucleostemin expression could not be specific in a certain type of cells in colon cancer cell line HT29 and controlling strategies in colon cancer must not be focused on one certain type of colon cancer cells as main expressing nucleostemin gene.
Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran.
Noosha
N
Zia-Jahromi
Seyed Hossein
SH
Hejazi
Mojtaba
M
Panjepour
Kazem
K
Parivar
Marjan
M
Gharagozloo
D000071916:AC133 Antigen; D015703:Antigens, CD / Q000235:genetics / Q000378:metabolism*; D003110:Colonic Neoplasms / Q000235:genetics / Q000378:metabolism*; D019204:GTP-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D006023:Glycoproteins / Q000235:genetics / Q000378:metabolism*; D019073:HT29 Cells; D006801:Humans; D016130:Immunophenotyping; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism*; D010455:Peptides / Q000235:genetics / Q000378:metabolism*
D000071916:AC133 Antigen; D015703:Antigens, CD; C469949:GNL3 protein, human; D006023:Glycoproteins; D009687:Nuclear Proteins; C000606023:PROM1 protein, human; D010455:Peptides; D019204:GTP-Binding Proteins