| ObjectivesPrimary aldosteronism is one of the most reason of symptomatic hypertension, The clinical manifestation of Primary aldosteronism is high blood pressure, hyperaldosteronemia,and hypokalemic. The most common clinical subtypes of primary aldosteronism are aldosterone—producing adrenocortical adenoma(APA)and adrenal cortical hyperplasia(or idiopathic hyperaldosteronism, IHA). The prevalence of APA may be as high as 60%-90%, but the pathogenesis of APA is still not clear. Current clinical diagnosis of PA is complicated, and part of APA ipatients after surgical removal of lesions still have hypertension.This technology have features of high sensitivity,high specificity,and high repeatability.We use to Illumina Infinium HumanMethylation27 BeadChip to analyze DNA methylation of Aldosterone-producing adanoma.Accroding to the result,using real-time PCR to detected the different expression of genes and analysis of Gene Function.Methods5 aldosterone-producing adenoma and 5 normal adrenal tissues were studied, which were confirmed by clinical and pathologic diagnosis. Infinium Methylation Assay were applied to analyze DNA methylation of Aldosterone-producing adanoma, using AVG beta represents to the level of methylation, Methylation scores represented as b values are generated for each site and are computed based on the ratio of methylated to methylated plus unmethylated signal outputs. Thus the b values range from 0 (unmethylated) to 1 (fully methylated) on a continuous scale.and Diffscore represent to the difference between aldosterone-producing adenoma and normal adrenal tissues,rang from-375-375;Secondly,9 aldosterone-producing adenoma and 6 normal adrenal tissues were studied, which were also confirmed by clinical and pathologic diagnosis. According to the results selecting AVG beta in APA group higher than the control group and the highest limits of Diffscore 4 genes, respectively BECN1, ATP2B4, ACAT1, MCCC1, using real-time PCR technology to verify the expression of these genes, make suer if Methylation is affected their expression.ResultsIn our experiment, according to 100≤Diffscore≤375, Adrenocortical adenoma compared with normal adrenal screened 2622 genes methylation is hypermethylated and 6179 gene is hypomethylated.it is show a similar pattern with a high peak of hypomethylated loci and a low peak in the hypermethylated loci.This is the expected U shape of methylation pattern of the genome but due to the array design which is skewed towards promoter region CpGs in CGIs shows the reverse pattern with a larger peak at the low end of the distribution.Among the hypermethylated gene, according to the AVG beta≥70%, Diffscore≥375,there are 8 genes; data analysis by computer and literature search, hypermethylated genes mainly related to the function is to regulate the cell cycle, inhibition of tumor growth, and promote protein phosphorylation, reduced aerobic metabolism; hypomethylated methylation mainly involves to the function is to promote tumor growth, regulating cell adhesion and promote tumor metastasis, stress response, increased endocytosis and defense response to protozoan; in the verification of the six genes, the expression of ATP2B4 is no significant difference between two group, BECN1, ACAT1, MCCC1 is downregulation and the difference is statistically significant, PPMIE, CCNT2 is upregulation and the difference is statistically significance. According to functional to classify genes, including tumor suppressor genes, metabolism related enzymes, including reduced protein phosphatase gene and cell cycle regulatory proteins.Conclusions:APA pathogenesis is still unknown. Gene chip can be used effiectively to find out abnormal gene methylation, and to f verify the expression changes, The genes that screened are decide by analysis, it is very useful for us of understanding the function of these genes. It is surporting that APA has Tumor characteristics, So they have very important significance of investigating the happening and development of APA. |
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