The Study Of APC、P53、K-ras And PIK3CA Gene Mutations In Colorectal Cancer Of China

Objectives: Colorectal cancer (Colorectal cancer, CRC), is one of the most commonmalignant tumor, its incidence and mortality rates are highest in the ranking of thedisease before five. Early diagnosis is an effective means to improve colorectal cancersurvival rates, but there is no convenient and sensitive diagnostic methods. The study ofgenetic changes is one of the colorectal cancer diagnosis and treatment based onresearch. This article aims to APC, P53, K-ras and PIK3CA gene mutations in theapplication of PCR and DNA sequencing in human colorectal cancer, and analyze theircorrelation with clinicopathological parameters, and study of these four genes that mayexist between the relationship between, for the process of colorectal tumorigenesis.Method: In this study, using whole genomic DNA target fragment was amplified byPCR and DNA sequencing technology in surgical specimens of79patients withcolorectal cancer, APC, P53, K-ras and PIK3CA gene mutation detection.①Use ofDNA Purification kit (Qiagen) to extract genomic DNA, and DNA quantitative UVspectrophotometer.②According to the classic literature, the NCBI’s Jiyinkuji for thecorresponding gene sequence information and primer5primer design software primerdesign.③using the corresponding primers worked out the right conditions, APC, P53,K-ras and PIK3CA four gene PCR.4Electrophoresis identification of PCRamplification by the specific product whether it had the size to meet the expected. The⑤qualified the product sent to the sequencing of Sanger sequencing.⑥sequencingcompany sent back the results were analyzed using Chromas software to read thesequencing map and the base sequence and confirm the corresponding sites of the corresponding amino acid coding sequence, and ultimately determine each specimenpoint mutation.⑦calculate the mutation frequency of each gene and to analyze themutation distribution characteristics.⑧each gene mutations and clinicopathologicalparameters (gender, age, lymph node metastasis, pathological stage and grading)correlation analysis and analysis of the correlation between the four-gene mutation inorder to explore these gene mutations in colorectal cancer diagnosis or treatment hasclinical significance.Results: The experimental results show that the APC gene mutation rate of25/79(31.6%), mutations in the distribution of performance-intensive for the region, thetypical mutation hotspot; P53gene mutation rate of25/79(31.6%), mutation hotspotcodon175(3/24),245(2/24),248(1/24),282(6/24); has the highest frequency ofK-ras gene mutation,30/79(38.0%), mutation hot spots located outside theobviouscodon on the sub-112,13; of PIK3CA gene mutations were7/79(8.9%),mutation hotspot codon542,545,546,1047. Four groups of gene mutation on the APC,P53, K-ras and PIK3CA correlation with clinicopathological parameters for statisticalanalysis, were not found statistically significant results of the analysis;4gene mutationpositive data between correlation analysis has not found its obvious relationshipbetween mutation. Tip4gene mutation has a catalytic role in the development ofcolorectal cancer. The results suggest that there is a considerable proportion of the APCin human colorectal cancer, P53, K-ras and PIK3CA gene mutation frequency, and doesnot have a significant positive and negative correlation between the mutation cases.Conclusion:①APC, p53, K-ras gene mutation frequency in human colorectal cancer,the APC gene mutation rate of25/79(31.6%), P53gene mutation rate of25/79(31.6%),K-ras gene mutation with the highestfrequency of30/79(38.0%), gene mutation hotspotmutations intensive regional distribution consistent with the Western populationreported.②Results of PIK3CA gene mutations in rectal cancer less frequently in theChinese population of PIK3CA gene mutations were7/79(8.9%), PIK3CA genemutation frequency differences between the Chinese population in Western populations.③Clinicopathological parameters of the four groups of genes APC, p53, K-ras andPIK3CA mutations were associated with age, gender, pathological stage, lymph node metastasis and tumor infiltration depth correlation analysis we found a significantcorrelationresults.④The analysis of the linkages between the four sets of mutations incases nor has obvious positive and negative, suggesting that this the APC, p53, K-rasand of PIK3CA four gene mutations for the development of colorectal cancer has acatalytic role, whichany genetic changes can lead to the occurrence and development ofcolorectal cancer.