TITLE:
CHK21100delC, I157T, IVS2 +IG > A, BRCA1 and BRCA2 Mutation Analysis in JF305: A Pancreatic Cancer Cell Line
AUTHORS:
Charles Ndawula Jr., Xueli Yang, Xiaohai Gong, Jian Jin
KEYWORDS:
BRCA1; BRCA2; CHK21100delC; CHK2I157T; CHK2IVS2 +IG > A; Homologous Mechanism; PARP; Brca1; Brca2
JOURNAL NAME:
Journal of Biosciences and Medicines,
Vol.2 No.1,
March
21,
2014
ABSTRACT: JF305 is a highly prolific pancreatic cancer cell line that originated from a Chinese patient. The cell line bears a functional HR double strand DNA repair mechanism but very responsive to PARP treatment a phenomenon clearly suggesting presence of an anomaly in the mechanism. Brca1, Brca2 and CHK2 proteins are very important constituents of the HR mechanism whose respective gene coding mutations are strongly associated with several cancers and are widely exploited in anticancer chemotherapy. In this current study, the BRCA1, BRCA2 gene mutation status in JF305 was determined together with the presence of 3 widely reported cancer linked CHK2 founder mutations (1100delC, I157T, IVS2 +IG > A). CHK21100delC genotype was determined using allele specific PCR, while the PCR-RFLP assay was used for I157T, IVS2 +IG > A analysis. PCR and direct sequencing were used for assessing the BRCA1 and BRCA2 gene. Results revealed that JF305 is CHK21100delC heterozygous mutant, CHK2I157T and CHK2IVS2 +IG > A wild type. Furthermore, it was observed that JF305 lacked BRCA1 and BRCA2 gene mutations. The mutation status identification of CHK2 and BRCA1/2 in JF305 provides a major milestone towards elucidating the properties of the cell line which subsequently promises to be an excellent model for evaluating the role of parp inhibitors in pancreatic cancer chemotherapy most especially in the respective cancer cell lines without BRCA1 and BRCA2 gene mutations.