Detection of p53 gene mutations by using PCR-RFLP method in cases of colon cancer

Colon cancer is a common disease that can be sporadic or inherited. p53 is the mostimportant tumor suppressor gene that is effective in cancer formation. Normal p53acts by causing G1 cell-cycle arrest to facilitate DNA repair during replication or toinduce apoptosis (programmed cell death). Arg72Pro may play a role in the earlystages of colorectal neoplasia and possibly in progression to invasive disease,depending on site and sex. Nucleotide alterations in p53 intron 6 have been reportedto be associated with the dysregulation of p53 function and tumor development.G13964C base change functioned as dominant mutation similar to the more commonmissense, nonsense and splice-site mutations.IIn this study, DNA was isolated from colon cancer tissue samples of (20 females and15 males) 35 cases diagnosed to be colon carcinoma. Tissues were collected from 35cases with colon cancer ages of female 40-90 (mean age 65.95±2.76) and ages ofmale 40-79 (mean age 61.67±3.07). Genomic DNA was extracted from paraffinembedded tissues using EZNA Tissue DNA Kit (Omega) following the protocol. Todetect the Arg72Pro region (exon 4) and G13964C variant (intron 6) PCR-RFLPassay was used. After then 4th exon of p53 were amplified by PCR method and p53mutation analyses was performed by using BstFNI restriction enzyme determined for4th exon. After then 6th variant of p53 were amplified by PCR method and p53mutation analyses was performed by using BstHHI restriction enzyme determined for6th intron. In 2 (%7.4) cases Pro/Pro, in 14 (%51.9) cases Arg/Arg and in 11 (% 40.7)Arg/Pro were found for Arg72Pro polymorphism. In 7 (%23.3) cases G/G, in 21(%70.0) cases G/C and in 2 (%6.7) C/C were found for G13964C polymorphism2006, 126 pageKey Words: Colon Cancer, p53 gene, Ar72Pro, G13964C, PCR, RFLP, mutation,polymorphism.II