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Objective:To explore the changes of mRNA N6-methyladenosine methylation level and methyltransferase-like 3 (METTL3) and demethylase fat mass and obesity-associated (FTO) in the blood of patients with Alzheimer's disease (AD) compared with normal controls.Methods:From January 2020 to June 2021, totally 40 AD patients treated in the outpatient and inpatient department of Neurology of the Affiliated Hospital of Jining Medical University were selected as the patient group, and 40 healthy volunteers as the control group. The blood samples were collected to extract plasma and peripheral blood mononuclear cells for enzyme-linked immunosorbent assay (ELISA), Western blot (WB), quantitative real-time PCR (qPCR) and m6A methylation quantification experiments respectively to detect the methylation levels of METTL3, FTO and m6A. The data were analyzed by SPSS 23.0 statistical software for t-test. Results:The plasma concentrations of METTL3 and FTO protein in AD group were lower than those in control group (METTL3: (22.33±3.01)ng/mL, (25.63±1.70)ng/mL, t=6.055, P<0.01; FTO: (63.51±4.95)pg/mL, (69.60±4.60)pg/mL, ( t=5.704, P<0.01). The band gray values of METTL3 and FTO protein in blood cells in AD group were lower than those in control group (METTL3: 0.399 5±0.028 7, 0.676 6±0.053 3, t=7.935, P=0.001; FTO: 0.439 4±0.017 8, 0.782 6±0.087 6, t=6.652, P=0.003). The expression levels of METTL3 and FTO in blood cell RNA in AD group were lower than those in control group (METTL3: 0.387 8±0.020 3, 1.010 0±0.177 0, t=6.041, P=0.004; FTO: 0.442 8±0.037 1, 1.003 0±0.090 4, t=9.931, P=0.001). The levels of m6A in blood cell RNA in AD group were lower than those in control group((0.000 571±0.000 167)%, (0.002 514±0.001 284)%, t=6.041, P=0.004). Conclusion:The levels of METL3, FTO and m6A methylation are down-regulated in the plasma and peripheral blood mononuclear cells of patients with AD, indicating that there is a certain association between mRNA N6-methyladenosine methylation and AD.
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Objective To evaluate the correlation of maternal FTO expression and insulin secretion levels in pregnant women.Methods In outpatient of Zhuhai Municipal Maternal and Child Healthcare Hospital,60 cases of pregnant women with gestational diabetes mellitus (GDM) and 60 cases of healthy pregnant women were recruited in this study from March 2016 to August 2016.Each pregnant woman was taken two tubes of venous blood.Lymphocytes was isolated from one tube of anticoagulated whole blood.The expression of FTO mRNA in lymphocytes was investigated using RT-PCR.Serum was isolated from another tube of non-anticoagulant whole blood.Its FTO protein was investigated using ELISA,insulin was investigated using chemiluminescence method,and GLU,CHO,TG,HDL-C and LDL-C were investigated by automatic biochemical analyzer.The correlation of the biochemical indicators were analyzed.Results The expression level of FTO mRNA,FTO protein,fasting glucose data,OGTT at 1 h and 2h glucose levels in the case group were 0.17 ％ ± 0.10 ％,61.32 ± 25.23 pg/ml,4.71 ±0.54 mmol/L,10.70±1.36 mmol/L and 8.97 ± 1.60 mmol/L respectively.The values of these biochemical indicators in control group were 0.11％±0.07％,51.47±22.97 pg/ml,4.41± 0.28 mmol/L,8.05±1.04 mmol/L and 6.56±0.75 mmol/L respectively.The differences were statistically significant (t =3.876,2.236,3.817,11.964 and 10.578,P＜0.05).The insulin levels were 6.83±9.76 mU/L in the case group,and 13.15±13.99 mU/L in control group.The differ ences was statistically significant too(t=-2.869,P＜0.05).FTO expression level was positively correlated with FTO protein,OGTT 1h glucose and OGTT 2h glucose levels (r=0.232,0.292,0.242,all P＜0.05),and it was negatively correlated with insulin levels(r=-0.185,P＜0.05).There was no correlation between FTO and fasting glucose expression levels (P ＞0.05).Conclusion The expression of FTO level in pregnant women with GDM was higher than that in healthy pregnant women.The relative expression of FTO mRNA was negatively correlated with insulin.And the regulation of glucose metabolism might be effected by insulin.
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Objective To evaluate the effect of Fat Mass and Obesity Associated (FTO) gene on radiosensitivity of human glioma cell A172 and investigate its potential mechanism by changing the expression of FTO gene.Methods Cells were divided into five groups according to their FTO protein expression level.The normal expression group was recorded as A172 Group,the low-expression and its negative control group was A172/siRNA and A172/NC Group,and the over-expression and its negative control group was A172/FTO and A172/PC group.FTO protein expressions were assayed by Western blot in A172 Group after irradiation.Clonogenic assay was executed to evaluate the relationship between FTO gene and radiosensitivity.Immunofluorescence and Western blot assay were applied to detect the proteins of DNA damage and repair.Results FTO protein expression level in A172 Group was significantly related to the irradiation dose and the time post-irradiation.The radiosensitization ratio (SERD0) of A172/siRNA and A172/FTO group were 1.829 and 0.812 respectively.Not only the number of γ-H2AX foci increased (t =-21.884,P ＜ 0.05) in A172/siRNA 1 h post-irradiation but the decreases of p-p95/NBS1 and Ku80 proteins were also detected (t =24.731,23.293,P ＜ 0.05) together with the increase of Rad50 protein (t =3.140,P ＜ 0.05).But the expressions of these proteins in A172/FTO group were opposite to the above phenomenon (t =0.642,-8.364,26.829,P ＜ 0.05).Conclusions FTO gene is a radiation-resistant gene,which may because the regulation of FTO gene could alter the primary injury and DNA damage repair in the irradiated tumor cells.
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Objective To study the association of fat mass and obesity associated gene(FTO gene) and genetic onset mechanism of obesity in Chinese children.Methods Two hundred and one Chinese children with obesity in Beijing Children's Hospital Affiliated to Capital Medical University from Jan.to Sep.2010,were selected as research subjects,183 healthy adult blood donors were selected as normal controls.Mass Spectrometry techniques were used to study the distributions of the alleles and gene type of FTO in patients and controls.And the relationship between FTO gene polymorphism and obesity in Chinese children were studied.Results The distributions of 5 FTO gene polymorphisms (rs9939609A,rs8050136A,rs3751812T,rs1421085C,rs7193144C) in obesity patients and healthy controls had significant differences.And the Haplotype analysis showed that all of the single nucleotide polymorphisms(SNPs) were in linkage disequilibrium,and three out of six (CTGGTCTGG,TCTGCAAAA,CTGGCCTGG) had significant differences between obesity patients and healthy controls (P ＜ 0.05).Conclusions The gene polymorphisms of rs9939609,rs8050136,rs3751812,rs1421085,rs7193144 of FTO gene confer significant susceptibility to obesity in Chinese children.The haplotypes of CTGGTCTGG,TCTGCAAAA,CTGGCCTGG have significant differences between obesity patients and healthy controls.