The Association Of Akt Oncogene And The Pathogenic Of Congenital Heart Disease

Objective:The purpose of this study was to analyses the sequence of the exon1-4 (encoding the pleckstrin homologue (PH) domain) of Akt1 and Akt2 genes in 18 cases of patients with congenital heart diseases (CHD). We expect to supply new experimental evidence for the molecular mechanism of pathogenic of CHD.Methods:Lymphocytes were isolated from peripheral blood which were obtained from eighteen CHD patients before the cardiopulmonary bypass(CPB). DNA extraction was processed according to the manufacturer's protocol. Akt1 exon1-4 and Akt2 exon1-4 were PCR amplified using forward and reverse primer sequences. The purification of the PCR products and the sequencing were completed. Sequence analysis was based on Akt1 and Akt2 DNA sequences (GenBank access no. NM₀01014432 for Akt1, and NM₀01626 for Akt2). The Akt1 protein levels were determined by Western Blotting Analysis.Results:No mutation in coding sequence was identified in this group of patients. However, a total of five different SNPs were identified in Akt1, yet none was found in Akt2. These Akt1 SNPs and their frequency are: i) A19853G (rs2498797) present in 16 of the 18 cases (88.9%); ii) A15756T found in 13 of the 18 cases (72.2%) ; iii) A19115G identified in 16 of the 18 cases (89%); iv) C15674T displayed in 1of the 18 cases (6%) and, v) C19155T (rs17846822) shown in 1 of the 18 cases (6%). Of the five SNPs, the one of C15674T (rs3730358) was previously found related with schizophrenia and affected Akt1 protein levels. We therefore examined the Akt1 protein levels in lymphocytes from peripheral blood of some of the 18 patients with CHD. The results demonstrated that the Akt1 protein levels are low in the patient with SNP of C15674T (rs3730358).Conclution:The cardiovascular defects in Akt knockout mice have demonstrated that disruption of Akt1 signaling contribute abnormal heart formation.The reduced Akt1 protein levels mean decreased Akt1 signaling which may compromise heart development. Here our study with CHD patients implicates that Akt1 may play a role in the pathogenesis of congenital heart diseases.