Induction of epigenetic reprogramming in fibroblast by extracts of carcinoma

Recently, differentiated somatic cells had been reprogrammed to pluripotential state in vitro, and various tissue cells had been elicited from those cells. Epigenetic modifications allow differentiated cells to perpetuate the molecular memory to retain their identity. In this study, NIH/3T3 fibroblasts were dedifferentiated with F9 cellular extract, and were detected using the gene expression pattern and the DNA methylation level in the dedifferentiated cells. The expression of embryonic marker Sox2, klf4, c-Myc and Oct4 were analyzed by real-time polymerase chain reaction (RT-PCR), and the total levels of cellular DNA methylation in dedifferentiated NIH/3T3 fibroblasts were measured by flow cytometry. According to the results, Sox2, klf4, c-Myc and Oct4 were expressed in NIH/3T3 fibroblasts after F9 cellular extract co-incubation. Also, the total DNA methylation level significantly decreased after the treatment. The data indicated that the extract of F9 cells can dedifferentiate NIH/3T3 cells, and make modified NIH/3T3 cells express transcription factors associated with totipotent and pluripotent cells.    Key words: Fibroblasts, reprogramming, methylation, epigenesis, carcinoma.