Hypomethylation Of PTEN Gene In Uyghur Patients With Mild Type 2 Diabetes Mellitus Leads To Insulin Resistance

Part One Expression of PTEN gene in peripheral blood mononuclear cells of Uyghur mild T2DM in Xinjiang Objectives: Insulin resistance is the central link of T2DM,and it is consistent throughout the course of T2DM.Phosphatase and tensin homologue on chromosome 10?PTEN?may play an important role in the mechanism of insulin resistance and may be a new target for improving insulin resistance.This study analyzed and compared the expression of PTEN gene in peripheral blood mononuclear cells?PBMC?of Xinjiang Uyghur first-episode T2DM and control group,and explored the relationship between PTEN abnormal expression and Xinjiang Uyghur T2DM.Methods: In this study,qRT-PCR and Western blot were used to detect the level of mRNA and protein expression of PTEN gene in peripheral blood mononuclear cells?PBMC?of Xinjiang Uyghur first-episode T2DM and control group.Results: The mRNA level of PTEN gene in mild T2DM patients in Xinjiang Uyghur was significantly higher than that in the control group.The relative transcript level of PTEN in the control sample was 1 and the relative transcript level in the Uighur T2DM sample was 3.5?min = 0.000211,max = 0.000879,SD: 0.0001765,t-test,P < 0.05?.The PTEN gene protein level in Uyghur patients with T2DM was significantly higher than that in the control group.?NGT: 0.67 ± 0.29,T2DM: 0.88 ± 0.40,t test,P = 0.0041?,while AKT protein levels were significantly reduced?t test,P = 0.013?.Our study found that low-density lipoprotein cholesterol?LDL-C?and Triglyceride and the protein expression level of PTEN showed a positive correlation trend,and there was a statistically significant difference.There was no significant correlation between the transcriptional levels of PTEN in other clinical features.Conclusions: This part of the study suggests that up-regulation of PTEN expression can lead to IR in Uyghur mild T2DM.Part two Methylation level of PTEN gene in peripheral blood mononuclear cells of Uyghur mild T2 DM in XinjiangObjectives: Epigenetics refers to the genetic mechanism that controls gene expression without affecting gene sequences in gene expression regulation.DNA methylation is one of the earliest discovered genetic modifications and is also well known.This study analyzed and compared the methylation status of PTEN gene in peripheral blood mononuclear cells?PBMC?of Xinjiang Uyghur first-episode T2 DM and control group,and screened for methylation site of T2 DM biomarker to explore PTEN promoter.The relationship between aberrant methylation and Xinjiang Uyghur T2 DM.The first part of the study was integrated to analyze the relationship between PTEN gene expression and DNA methylation in its promoter region.Methods: This section uses the high-throughput analysis of Mass ARRAY technology to compare the overall and individual Cp G methylation levels of the PTEN gene promoter region in the Xinjiang Uyghur mild T2 DM and control PBMC.Results:?1?We selected methylation levels relative to the PTEN gene translation initiation site from-2,515 bp to-2,186 bp.Check the 330 bp area using the Mass ARRAY platform.The PTEN promoter region contains 21 Cp G sites,which we divided into 15 Cp G units.Of these units,3 Cp G units?6 Cp G sites?did not successfully obtain the test results.The final data set consisted of 12 Cp G units from 94 samples.?2?The average global DNA methylation level of the Uyghur mild T2 DM samples ranged from 2.08% to 5.58%,and the average DNA methylation frequency ranged from 4.17% to 11.25% in the control samples.Abnormal methylation of PTEN in Uighur T2 DM samples was significantly lower than that of NGT samples,with an average of 3.27% and 7.28%,respectively?t test,P < 0.0001?.In addition,there is no correlation between the abnormal methylation level of PTEN and clinical indicators.In addition,we performed a two-way hierarchical clustering analysis of Cp G unit methylation data of Uighur T2 DM and NGT samples.Significantly different methylation patterns were found.?3?There is some variability in the methylation of individual Cp G sites for different samples.The methylation levels of most Cp G units in Uighur T2 DM samples were numerically lower than those of NGT samples,with Cp G₉ and Cp G₂1 methylation levels significantly different between these groups?one-way ANOVA,P < 0.05?.?4?In the Uighur mild T2 DM and control groups,we determined a negative correlation between DNA methylation levels and PTEN transcript levels?95% CI: 0.00009339 to 0.00002474,P = 0.001,r = 0.45?.Linear regression analysis showed a significant correlation between PTEN DNA methylation levels and PTEN protein expression?two-tailed Pearson test,95% CI: 0.1897 to 0.03355,P = 0.006,r = 0.38?.?5?There was no significant correlation between Cp G₂1 methylation level and clinical features in mild T2 DM group.In the T2 DM group,most clinical features were not significantly associated with Cp G₉ methylation levels,but our study found a negative correlation between waist circumference and BMI and HOMA-IR and Cp G₉ methylation in diabetic patients.Conclusions:?1?In Uyghur patients with mild T2 DM,hypomethylation of PTEN may be responsible for the high expression of PTEN.The hypomethylation level of PTEN is accompanied by a decrease in phosphorylation of PI3K/AKT signaling pathway,which leads to IR;?2?The DNA methylation level of Cp G₉ site is related to IR-related phenotype,which has certain significance for revealing the mechanism of IR in Uyghur T2 DM.