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Yakhteh Medical Journal. 2010; 12 (2): 223-230
in Persian, English | IMEMR | ID: emr-98593

ABSTRACT

Evaluating the expression of Oct-4, NANOG, Sox2 and Nucleostemin in colon cancer cell lines [Caco-2 and HT-29]. Caco-2 and HT-29 human colon cancer cell lines were cultured in Dulbecco's modified eagles medium [DMEM] and Roswell Park Memorial Institute medium [RPMI] respectively, containing 10% fetal bovin serum [FBS] with 1% peniciline and streptomycinen in 37°, 5% CO2 incubator. Total RNA was isolated using the ISOGEN method. RNA integrity was checked with the use of agarose gel electrophoresis and spectrophotometry. Reverse transcriptase polymerase chain reaction [RT-PCR] was used to examin the samples. The expression of Oct-4 and Nucleostemin at the protein level was further determined using immunocytochemistry. RT-PCR analysis of Caco2 and HT-29 colon cancer cell lines showed expression of Oct-4, NANOG, Sox2 and Nucleostemin genes. Also immunocytochemical analysis confirmed the cytoplasmic and nuclear expression of the Oct-4 protein and Nucleostemin proteins. Collectively, our data confirmed the expression of Oct-4, NANOG, Sox2 and Nucleostemin in colon cancer cells and suggested that their expression can be used as potential tumor markers in diagnosis and /or prognosis of colon tumors. These results confirm the potential value of the cancer stem-cell theory in cancer therapy


Subject(s)
Humans , Neoplastic Stem Cells , Homeodomain Proteins , SOXB1 Transcription Factors , Nuclear Proteins , GTP-Binding Proteins , Cell Line , Spectrophotometry , RNA , Electrophoresis, Agar Gel , Reverse Transcriptase Polymerase Chain Reaction , Immunohistochemistry
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