بيان ژن هاي خودبازسازي Nanog, Nucleostemin,Oct-4 و Sox2 در رده هاي سلولي سرطان كولون Caco2 و HT-29

Evaluating the Expression of Oct-4, NANOG, Sox2 and Nucleostemin in Colon Cancer Cell Lines (Caco-2 and HT-29)

Objective: Evaluating the expression of Oct-4, NANOG, Sox2 and Nucleostemin in colon cancer cell lines (Caco-2 and HT-29). Materials and Methods: Caco-2 and HT-29 human colon cancer cell lines were cultured in Dulbeccoʹs 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% C02 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. Results: 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. Conclusion: 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.