Single nucleotide variations in cultured cancer cells: effect of mismatch repair

Publication date: Available online 27 July 2017 Source:Mutation Research/Fundamental and Molecular Mechanisms of Mutagenesis Author(s): Igor G. Panyutin, Irina V. Panyutin, Ian Powell-Castilla, Laura Felix, Ronald D. Neumann We assessed single nucleotide variations (SNVs) between individual cells in two cancer cell lines; DU145, from brain metastasis of prostate tumor with deficient mismatch repair; and HT1080, a fibrosarcoma cell line. Clones of individual cells were isolated, and sequenced using Ion Ampliseq comprehensive cancer panel that covered the exomes of 409 oncogenes and tumor suppressor genes. Five clones of DU145 and four clones of HT1080 cells were analyzed. We found from 7 to 12 unique SNVs between DU145 clones, while HT1080 clones showed no more than one unique SNV. We then sub-cloned individual cells from some of these isolated clones of DU145 and HT1080 cells. The sub-clones were expanded from a single cell to approximately one million cells after about 20 cell divisions. The sub-clones of DU145 cells had from one to four new unique SNVs within the sequenced regions. No unique SNVs were found between sub-clones of HT1080 cells. Our data demonstrate that the extent of genetic variation at the single nucleotide level in cultured cancer cells is significantly affected by the status of the DNA mismatch repair system. Graphical abstract

Source: Mutation Research Fundamental and Molecular Mechanisms of Mutagenesis - July 27, 2017 Category: Cytology Source Type: research