Transfection of the H-RAS gene produces genomic instability in the MOLT-4 cell line

Objective:The aim of this study was to evaluate the effect of the normal and mutated H-RAS gene on genomic instability and apoptosis in MOLT-4 cells.Material and methods: The MOLT-4 cells were cultured and DNA was extracted.Then, the region that compromises both codons (12 and 13) of the exon 1 in the H-RAS gene was amplified by PCR and sequenced.The pSV2neo, pSV2neo-Ras wt , and pSV2neo-Ras mut plasmids were extracted from the DH5-alpha strain, sequenced, and transfected in MOLT-4 cells.Finally, the number of chromosomes was analyzed by means of karyotype and the number of apoptotic bodies was evaluated using acridine orange with propidium iodide.Results: No mutations were found on the H-RAS gene in the region of the 12 and 13 codons of exon 1 in the MOLT-4 cells.On the other hand, the sequences of the pSV2neo-Ras wt and pSV2neo-Ras mut plasmids matched with the normal and mutated versions of H-RAS, respectively.An increase in the number of chromosomes per cell with the normal and mutated H-RAS genes was also observed, as well as, the presence of apoptotic bodies.Conclusions: There are no H-RAS gene mutations in the 12 and 13 codons of exon 1 in MOLT-4 cells.The pSV2neo-Ras wt and pSV2neo-Ras mut plasmids contain normal and mutated versions of H-RAS, respectively.The normal and mutated H-RAS genes induce genomic instability and apoptosis in MOLT-4 cells.Plasmids pSV2neo-Ras wt and pSV2neo-Ras mut contain normal and mutated versions of the H-RAS gene.Normal and mutated H-RAS genes induce genomic instability and apoptosis in MOLT-4 cells.