The Study On Relationship Between HLA-DQB1~*03 Alleles And Cervical Cancer

ObjectiveTo investigate the relationship and degree between HLA-DQB1~*03 alleles(0301,0302,0303,0304)including six heterozygote types and the development of CIN,cervical cancer in population of Tianjin area in order to find pathogenic genetic base of cervical cancer and precancerous lesion,furthermore provide new experimental reference for clinic screening out cervical cancer and precancerous lesion or it may become the checking method of cervical cancer susceptible populations in epidemiology.MethodsExperimental objects were grouped into three teams:invasive carcinoma of cervix uteri team,CIN team and normal controls team.DNA was extracted by phenol-chloroform-amylene alcohol method from the peripheral vein blood of experimental objects.The target DNA fragments of HLA-DQB1~*03 alleles were amplified by using polymerase chain reaction of sequence specific primers (PCR-SSP).PCR productions were checked by agarose gel electrophoresis,if there is special band,it will be sequenced and compared with 2th exon sequence of HLA-DQB1~*03 alleles in order to verify the accurance of this experimental method. Chi-square test of frequency distributions under completely randomized design was Conducted by SAS 9.0 software and frequency distributions of DQB1~*03 alleles including six heterozygote types among three teams were compared.Frequency distributions of DQB1~*0301,0302,0303 multiple alleles were compared also.In addition,the clinical information including the incidence of postcoital vaginal bleeding,the stages,the incidences of pelvic lymph nodes metastasis and pathological type of 52 cases of the cervical cancer was analysed retrospectively.Results 1.PCR productions were separated by agarose gel electrophoresis,special band appeared in corresponding swimming road.PCR productions were sequenced,the sequences were uniform.2.The frequency distributions of DQB1~*0301,0302,0303 multiple alleles were significantly higher in CIN team,invasive carcinoma of cervix uteri team than that in normal controls team,that is to say,frequency distributions of DQB1~*0301,0302,0303 multiple alleles among three teams has statistic significance(p＜0.05),but DQB1~*0304 has no statistic significance(p＞0.05). Six heterozygote types were permuted and combined by HLA-DQB1~*0301-04 multiple alleles:DQB1~*0301/DQB1~*0302,DQB1~*0301/DQB1~*0303,DQB1~*0301/DQB1~*0304,DQB1~*0302/DQB1~*0303,DQB1~*0302/DQB1~*0304 and DQB1~*0303/DQB1~*0304.Only DQB1~*0301/DQB1~*0303 has statistic significance(p＜0.05).3.Frequency distributions of HLA-DQB1~*0301,0302 and 0303 multiple alleles has statistic significance(p＜0.05),DQB1~*0301 and 0303 multiple alleles were higher than DQB1*0302.4.The incidence of postcoital vaginal bleeding was lower in postcoital wemen than that in remenopausal wemen(p＜0.01);The incidence of cervical adeocarcinomai was lower in postmenopausal wemen than that in prmenoposal wemen(p＜0.05).ConclusionsSequencing analyse may validate the accurance of PCR-SSP for screening out HLA-DQB1~*03 alleles.There may be positive correlation between HLA-DQB1~*0301,0302,0303 and DQB1~*0301/DQB1~*0303 heterozygote type and the development of CIN and cervical cancer.The cervical cancer screening is thought to be necessary in postmenopausal wemen.