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ObjectiveTo evaluate the relationship and significant between low molecular-weight protein (LMP) methylation and expression,and Uyghur women cervical lesions with human papillomavirus (HPV) infection.MethodsGenetic information was obtained from the GenBank database,specialized software was used to scan gene promoter regions,and CpG island fragment specific primers was designed,gene methylation and CpG site sequences related information were gained by the PCR amplification,vector cloning and sequencing of the bisulfate-modified cervical cancer cell DNA.Methylation status of LMP was quantitative evaluated by Sequenom MassARRAY DNA in 78 subjects with different cervical lesions; mRNA and protein of LMP2 and LMP7 were evaluated by RT-PCR and immunohistochemistry.HPV infection status determined use HPV gene chips.ResultsGene promoter region CpG fragments methylation sequencing it was detected that selected CpG sites of cervical cancer cell genome LMP7 had methylation.However no methylation site was found in gene promoter region of LMP2.The percent of LMP7 methyiation was increased steadily with the severity of cervical lesions.showing 0.0652±0.0488,0.0728+0.0548 and 0.1864+0.0893 of which with normal control,CIN and CSCC (P＜0.01).LMP7 was significantly reduced in CSCC and CIN compared with normal cervical epithelium,and its mRNA expression consistent with the results of proteins,while no significant difference in LMP2 expression.Moreover,the methylated proportion of LMP7 was negatively associated with the protein expression in cervical lesions ( F =8.69,P =0.035 ).Stratified analysis indicated that the percents of LMP7 methylation in subjects with HPV16 positive still increased( t=1.996,P=0.049).ConclusionOur findings indicated that LMP7 methylation was significantly associated with cervical lesions and HPV infection.
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<p><b>BACKGROUND</b>It has been confirmed that defective expression of human leukocyte antigen class I (HLA-I) molecules can contribute to the immune evasion of cancer cells in some types of cancer. The aim of this study was to examine the expression of HLA class I antigen and the antigen-processing machinery (APM) components in esophageal squamous cell carcinoma (ESCC) and their role in high risk human papillomavirus (HPV) infection, and to analyze their association with histopathological characteristics in the Kazak ethnic group.</p><p><b>METHODS</b>A total of 50 formalin-fixed, paraffin-embedded ESCC lesions were collected from the First Affiliated Hospital of Xinjiang Medical University, China. The expression levels of HLA-I antigen and APM components were determined by immunohistochemistry; the HPV DNA were detected using polymerase chain reaction (PCR).</p><p><b>RESULTS</b>A high frequency of down-regulation or loss of expression of HLA and APM components were found in esophageal cancer in Kazak people. HLA-I, TAP1, CNX, LMP7, Erp57, Tapasin and ERAP1 were down-regulated in 68%, 44%, 48%, 40%, 52%, 32% and 20% of ESCC lesions then, respectively. The loss of expression of HLA-I antigen was significantly correlated with part of the APM components and positively correlated with high risk HPV16 infection. TAP1, CNX, LMP7, Erp57 and Tapasin loss were significantly associated with tumor grading, lymph node metastasis and depth of invasion (P < 0.05).</p><p><b>CONCLUSION</b>Our results suggest that APM component defects are a mechanism underlying HLA-I antigen down-regulation in ESCC lesions, and indicate that the loss expression of HLA-I and APM components will become an important marker of ESCC and analysis of HLA-I and APM component expression can provide useful prognostic information for patients with ESCC from the Kazak ethnic group.</p>
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , ATP Binding Cassette Transporter, Subfamily B, Member 2 , ATP-Binding Cassette Transporters , Genetics , Metabolism , Aminopeptidases , Genetics , Metabolism , Antigen Presentation , Genetics , Physiology , Calnexin , Genetics , Metabolism , Esophageal Neoplasms , Metabolism , Histocompatibility Antigens Class I , Genetics , Metabolism , Human papillomavirus 16 , Genetics , Immunohistochemistry , In Vitro Techniques , Membrane Transport Proteins , Genetics , Metabolism , Minor Histocompatibility Antigens , Polymerase Chain Reaction , Proteasome Endopeptidase Complex , Genetics , Metabolism , Protein Disulfide-Isomerases , Genetics , MetabolismABSTRACT
Objective To study metabonomics in the urine of rats of different genders by magnetic resonance (MR) with 2 normalization methods. Methods Different normalization methods such as mean-centering not scaling (Ctr) and unit variance scaling (UV) were used before orthogonal to partial least squares discriminant analysis(OPLS-DA). Distinguished metabolites in the urine of different gender rats were analyzed by calculating the correlation coefficients. Results The data normalized by Ctr before OPLS-DA analysis revealed high degree conception metabolites in the urine such as valine, alanine, acetate, ornithine, aminohippurate, phenylethylamine, cytosine, citrate, dimethylamine, allantoin, methylamine, fumarate and one unknown metabolite whose chemical shift was δ4.14. Data normalized by UV before OPLS-DA analysis revealed the above 12 high degree conception metabolites except citrate, and also low degree conception metabolites such as thiamine, creatinine, formate and one unknown metabolite whose chemical shift was δ2.92.Conclusion Unit variance scaling is a more effective normalization method in metabonomic analysis.
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<p><b>OBJECTIVE</b>To analyze the metabonomic (1)H-MRS of plasma samples from patients with esophageal cancer and healthy controls applying different pattern recognition methods, and to explore the potential of application of (1)H-MR-based metabonomics in clinical research.</p><p><b>METHODS</b>(1)H-MR was performed on plasma samples from 109 EC patients and 50 health controls to analyze the metabonomic variation between EC patients and healthy subjects and the corresponding (1)H-MRS were recorded on Varian Unity ANOVA 600 MHz to perform principal components analysis (PCA), partial least squares discriminant analysis (PLS-DA), orthogonal partial least squares discriminant analysis (OPLS-DA), respectively.</p><p><b>RESULTS</b>OPLS-DA analysis could correctly separate almost all the plasma samples from EC patients and health controls, better than both the PCA and PLS-DA. The plasma levels of leucine, alanine, isoleucine, valine, glycoprotein, lactate, acetone, acetate, choline, isobutyrate, unsaturated lipid, VLDL, LDL, 1-methylhistidine were significantly decreased in EC patients (r total > 0.27, P < 0.05), while that of dimethylamine, α-glucose, β-glucose, citric acid increased in the EC patients (r total < -0.27, P < 0.05).</p><p><b>CONCLUSIONS</b>The analysis of metabonomic (1)H-MRS of plasma samples by OPLS-DA method can eliminate the influence of non-experimental factors and decrease the heterogeneity of samples. It is useful and of great potential for application in clinical diagnosis and research of esophageal cancer.</p>