Choosing the right cell line for rectal cancer research
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AuthorMoya Ramírez, Miguel Ángel; Sánchez Martín, Victoria; Herrera Merchán, Antonio; Medina Vico, Pedro Pablo; Cuadros Celorrio, Marta Eugenia
Archivos de Medicina Universitaria
C-MYCRectal tumorResponseCell line
Moya Ramírez, Miguel Ángel; Sánchez Martín, Victoria; Herrera Merchán, Antonio; Medina Vico, Pedro Pablo; Cuadros Celorrio, Marta Eugenia. Choosing the right cell line for rectal cancer research. AMU. 2015; 3: 17-20
Up to date no effective method exists that predicts response to preoperative chemoradiation (CRT) in locally advanced rectal cancer (LARC). Nevertheless, identification of patients who have a higher likelihood of responding to preoperative CRT could be crucial in decreasing treatment morbidity and avoiding expensive and time-consuming treatments. Using the Gng4, c-Myc, Pola1, and Rrm1 signature, we were able to establish a model to predict response to CRT in rectal cancer with a sensitivity of 60% and 100% specificity. The aim of this study was to characterize c-Myc status in DNA, RNA and protein levels in 3 tumoral cell lines (SW480, SW620 and SW837) to establish the best cell line model and, subsequently, carry out genome silencing of c-Myc by means of RNA interference (iRNA). To study the expression levels of c-Myc, we used Polymerase Chain Reaction (PCR) amplifications and sequencing; quantitative real time PCR (qRT-PCR); and western blot analysis in each cell line. SW480 and SW620 showed a variation A > G in exon 2, which caused a substitution of aspargine to serine, and SW837 revealed a G > A transition in the same, which caused a mutation at codon 92. The three cell lines expressed c-Myc mRNA. SW837 showed a decrease of c-Myc expression levels compared with SW480, and SW620. At protein level, SW620 showed the highest expression of c-Myc. According to the results obtained, we can perform c-Myc gene silencing experiments to analyze the role of this biomarker in response to treatment.