ABSTRACT
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.