Analysis Of HPV16L1Gene Variants Of Different Cervical Lesions In Qingdao Abstract

ObjeetiveHuman papillomavirus (HPV) has a number of intratypic variants, with racial specificity and different geographical distribution.This study was performed to detect sequence variants of the HPV16late gene L1derived from Qingdao women with different cervical lesions,analyze the structural features and variation rule, discuss the association between the sequence variant and the cervical lesions, and to provide experiment foundation of the development of thigh specific vaccine.Materials and Methods(1)61cervical cancer biopsies.with were diagnosed squamous cervical cancer by pathology, and300cervical-scraping cell lysates (LCT) were collected in the Qingdao area. The LCT tissues include47normal tissues,100Lowgrade Squamous Intraepithelial Lesion (LSIL) tissues,and153Highgrade Squamous Intraepithelial Lesion (HSIL) tissues. In all LCT tissues.181tissues were pathologically diagnosed.including40chronic cervical inflammation and141cervical intraepithelial neoplasia (CIN) I～III.(2) DNA samples were extracted from the cervical scrapes and tissues.(3) DNA samples were amplified by polymerase chain reaction(PCR)、using specific primers MY09/11for HPV infection, specific HPV16primers to identify the positive tissues.(4) DNA samples extracted from the HPV16positive tissues were amplified by the primers for L1.(5) the PCR fragments of HPV16L1gene were sequenced and compared with the type stain by DNASTAR.Results (1) The positive rate of HPV was75.0%(30/40、89.4%(126/141) and93.4%(57/61) separately in inflammation. CIN and cervical cancer. The distribution showed statistical differences (X2=8.370, P=0.015). The positive rate of HPV was55.3%(26/47),61.0%(61/100)、81.7%(125/153)and93.4%(57/61)separately in normal cells, LSIL,HSIL and cervical cancer cells. The distribution showed statistical differences (X2=34.405, P=0.000)(2) The positive rate of HPV16was30.0%(12/40)、41.1%(58/141) and65.6%(40/61) separately in inflammation, CIN and cervical cancer. The distribution showed statistical differences (X2=14.874, P=0.001). The positive rate of HPV16was10.6%(5/47)、32.0%(32/100)、53.6%(82/153) and65.6%(40/61) separately in normal cells. LSIL,HSIL and cervical cancer cells. The distribution showed statistical differences (X2=44.303, P=0.000)(3) A total of96samples were amplified successfully in all samples diagnosed by pathology. The rate were25.0%(10/40)、37.6%(53/141) and54.1%(33/61) separately in inflammation. CIN and cervical cancer. In all LCT samples.136were amplified successfully, with70HSIL.28LSIL and5normal cells.(4) A total of13variants were found in the full length of L1gene. Five mutations, including C6240G、A6432G、6902insATC、6954delGAT and G7061A(nonsense). were found in all of the positive samples. Mutation of A6178C was another hot spot, whose distribution showed no significant difference among inflammation、CIN and cervical cancer (X2=1.265, P=0.531). and also no significant difference among normal cells. l.SIL,HSIL and cervical cancer cells(X2=6.705, P=0.082).Conclusions(1) The C6240G、A6432G、6902insATC、6954delGAT、G7061A(nonsense) and A6178C were the prevalent variants in Qingdao cervical lesions. There are significant geographical features in the HPV161.1gene variation. (2) The HPV16L1gene variation may cause certain changes concerning immunogenicity and antigenicity of the virus, nevertheless has no apparent connection to the development of cervical lesions.