Found mutations in codons 601 (p.Lys601Glu), 466 (p.Gly466Glu), and 471 (p.Val471Ala) of BRAF, the latter not previously described in mCRC [23]. A single BRAF mutation, p.Val471Ala, occurred inside a tumor also carrying a PIK3CA p.His1047Arg mutation.PIK3CAPIK3CA mutations had been present in 10.9 (22/201) on the tumors, unequally distributed among exonsGuedes et al. BMC Cancer 2013, 13:169 http://www.biomedcentral.com/14712407/13/Page four ofFigure 1 High resolution melting analysis of PIK3CA exon 9. A) Normalized and B) distinction graph, containing wildtype samples (blue) and mutated samples (green and red).9 and 20: 73 (16/22) have been helical domain mutants (p.Glu542Lys, p.Glu545Lys, p.Glu545Asp, and p.Gln546Lys) and 27 (6/22) kinase domain mutants (p.Met1043Ile, p.His1047Arg, p.His1047Leu, and p.His1047Tyr). Five from the PIK3CA mutants also contained yet another mutation in either KRAS or BRAF.Clinicopathological associationsKRAS mutations had been a lot more frequent in patients older than the median age of diagnosis (21.five vs. 8.two ; P=0.034), whereas no statistically important differences have been identified for BRAF or PIK3CA mutations regarding this parameter.BRAF and PIK3CA mutations were much more frequent within the colon than in the sigmoid or rectum: 20.eight vs. 1.6 vs. 0.0 (P=0.000) for BRAF and 23.four vs. 12.1 vs. 5.four (P=0.011) for PIK3CA mutations (Figure 3). Although the frequency of KRAS mutations is greater in sigmoid and rectum, the distinction is just not statistically substantial. No important differences have been found in between genders with regards to KRAS, BRAF, or PIK3CA mutation frequencies.76271-74-4 Data Sheet Discussion We here show that extra than onefourth of KRAS exon 2 wildtype mCRC patients present other mutations in KRAS,Guedes et al.Formula of 128625-52-5 BMC Cancer 2013, 13:169 http://www.PMID:36014399 biomedcentral.com/14712407/13/Page five ofTable 1 Frequency of KRAS, BRAF and PIK3CA single mutants (N=47/201)Mutation Gene and exon KRAS exon three cDNA sequence c.176_178del c.179GT c.181CA c.182AT c.182AG c.183AC KRAS exon 4 BRAF exon 11 BRAF exon 15 c.436GA c.1397GA c.1799 TA c.1808AG PIK3CA exon 9 c.1624GA c.1633GA c.1635GT c.1636CA PIK3CA exon 20 c.3129GA c.3140AG c.3140AT Protein sequence p.Ala59del p.Gly60Val p.Gln61Lys p.Gln61Leu p.Gln61Arg p.Gln61His p.Ala146Thr p.Gly466Glu pVal600Glu p.Lys601Glu p.Glu542Lys p.Glu545Lys p.Glu545Asp p.Gln546Lys p.Met1043Ile p.His1047Arg p.His1047Leu No. of Total no. of situations instances ( ) 1 1 1 1 3 2 1 10 1 eight 1 5 5 1 three 1 1 1 3 (1.5 ) 14 (7.0 ) ten (five.0 ) 1 (0.five ) 9 (four.5 ) ten (5.0 )c.151_195dup p.Cys51_Ser65dupBRAF, and/or PIK3CA and report 5 novel mutations, namely four in KRAS exon three and a single in BRAF exon 11. The mutational frequencies inside the most normally altered codons of KRAS, BRAF, and PIK3CA genes had been in accordance to those previously described in the literature. We’ve got previously shown that HRM is really a highly sensitive method to detect mutations in mCRC, being significantlyTable two Frequency of double mutants (N=6/201)Coexisting mutations Gene and exon PIK3CA exon 9 KRAS exon 4 PIK3CA exon 9 KRAS exon three PIK3CA exon 20 KRAS exon 3 PIK3CA exon 20 KRAS exon four PIK3CA exon 20 BRAF exon 11 KRAS exon three KRAS exon four cDNA sequence c.1633GA c.436GA c.1624GA c.183AC c.3139CT c.173_217dup c.3140AG c.436GA c.3140AG c.1412 TC c.145GA c.436GA Protein sequence p.Glu545Lys p.Ala146Thr p.Glu542Lys p.Gln61His p.His1047Tyr p.Thr58_Met72dup p. His1047Arg p.Ala146Thr p.His1047Arg p.Val471Ala p.Glu49Lys p.Ala146Thr 1 1 1 1 1 No. of cases 1 Total no. of situations ( ) 6 (three.0 )far more sensitive and less costly than.