Open AccessMurine melanoma cells incomplete reprogramming using non‐viral vector
Author(s) -
Câmara D.A.D.,
Porcacchia A.S.,
Costa A.S.,
Azevedo R.A.,
Kerkis I.
Publication year - 2017
Publication title -
cell proliferation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.647
H-Index - 74
eISSN - 1365-2184
pISSN - 0960-7722
DOI - 10.1111/cpr.12352
Subject(s) - reprogramming , homeobox protein nanog , sox2 , induced pluripotent stem cell , biology , transfection , lin28 , cell culture , viral vector , microbiology and biotechnology , stem cell , cancer cell , klf4 , cell , embryonic stem cell , gene , cancer , genetics , recombinant dna
Objectives The reprogramming of cancer cells into induced pluripotent stem cells or less aggressive cancer cells can provide a modern platform to study cancer‐related genes and their interactions with cell environment before and after reprogramming. Herein, we aimed to investigate the reprogramming capacity of murine melanoma B16F10 cells. Materials and methods The B16F10 was transfected using non‐viral circular DNA plasmid containing the genes Sox‐2, Oct4, Nanog, Lin28 and green fluorescent protein ( GFP ). These cells were characterized by immunofluorescence, analysis RT ‐ PCR and cell cycle. Results Our results demonstrated for the first time that reprogramming of B16F10 may be induced using non‐viral minicircle DNA containing the four reprogramming factors Oct4, Sox2, Lin 28, Nanog ( OSLN ) and the GFP reporter gene. The resulting clones are composed by epithelioid cells. These cells display characteristics of cancer stem cells, thus expressing pluripotent stem cell markers and dividing asymmetrically and symmetrically. Reprogrammed B16F10 cells did not form teratomas; however, they showed the suppression of tumourigenic abilities characterized by a reduced tumour size, when compared with parental B16F10 cell line. In contrast to parental cell line that showed accumulation of the cells in S phase of cell cycle, the cells of reprogrammed clones are accumulated in G1 phase. Long‐term cultivation of reprogrammed B16F10 cells induces regression of their reprogramming. Conclusions Our data imply that in result of reprogramming of B16F10 cells less aggressive M urine M elanoma R eprogrammed C ancer C ells may be obtained. These cells represent an interesting model to study mechanism of cells malignancy as well as provide a novel tool for anti‐cancer drugs screening.