Objective To establish a high quality genomic DNA preparation method from formalin fixed and paraffin embedded tissues (FFPET) by integrating previous DNA extraction methods. Methods We combined classical genomic DNA extraction methods and commercially available DNA affinity column, replaced the de-waxing by dimethylbenzene with water-bath, designed a fast and improved genomic DNA preparation method. We also extracted genomic DNA from paraffin embedded cervical cancer tissues, and checked the quality of DNA by agarose gel electrophoresis and polymerase chain reaction detection. Results The improved genomic DNA extraction method combined the advantages of the water-bath de-waxing and DNA affinity column, making it possible to get high quality genomic DNA from paraffin embedded cervical cancer tissues, and especially efficient to recover genomic DNA fragments larger than 20 kb. Conclusion The improved DNA extraction method is fast and convenient to recover high quality genomic DNA from paraffin embedded tissues.

OBJECTIVETo explore the relationship between human papillomavirus(HPV) infection and expression of human leukocyte antigen class I (HLA-I) family genes (HLA-A, B and C) in cervical cancers of Uighur women, and to investigate their effect on cervical cancer progression.

METHODSFresh tissue samples of 78 Uighur women with cervical squamous carcinoma, cervical intraepithelial neoplasia (CIN) or benign cervicitis were selected. HLA-A, B and C expression and HPV infection were analyzed using RT-PCR and HPV gene chips, respectively.

RESULTSThere was a tendency of increasing the total loss of HLA-A, B and C mRNA as the cervical lesions became more aggressive. Loss of HLA-I mRNA in CIN (I, II and III) and cervical squamous carcinoma was 70.0% (14/20) and 84.8% (39/46) respectively. Poorly differentiated cervical carcinomas had the highest HLA-I expression loss (90.6%). In contrast, HLA-I mRNA loss was seen in only 8% of cases of cervicitis. Moreover, it was found that high risk HPV 16 infection was strongly correlated with the loss HLA-I mRNA expression (r = 0.803, P < 0.01).

CONCLUSIONSThe loss of HLA-I gene expression is strongly correlated with HPV-16 infection, and may serve as a biomarker of cervical cancer progression in Uighur women.

Adult ; Aged ; Carcinoma, Squamous Cell ; ethnology ; genetics ; immunology ; virology ; Cervical Intraepithelial Neoplasia ; ethnology ; genetics ; immunology ; virology ; China ; ethnology ; Female ; HLA Antigens ; genetics ; metabolism ; HLA-A Antigens ; genetics ; metabolism ; HLA-B Antigens ; genetics ; metabolism ; HLA-C Antigens ; genetics ; metabolism ; Human papillomavirus 16 ; isolation & purification ; Humans ; Middle Aged ; Papillomavirus Infections ; ethnology ; genetics ; immunology ; virology ; RNA, Messenger ; metabolism ; Uterine Cervical Neoplasms ; ethnology ; genetics ; immunology ; virology ; Uterine Cervicitis ; ethnology ; genetics ; immunology ; virology