| Objective: To investigate the distributional characteristics of genetic variation of HPV 16 E6 in women health screenings of Tianjin urban areas and to analyse the genetic variant condition of HPV16, and study the association between of HPV 16 E6 variants and progression of cervical lesions, find out the high-risk variant among Tianjin, to provide a theoretical basis for the study of HPV16 highly carcinogenic capacity factors and to provide important epidemiological data for the development of HPV therapeutic vaccine.Methods: A cervical cancer screening study for 2000 intelligent women was developed from April to October, 2013 in Tianjin urban areas. All women have had sexual life. Every participant signed informed consent and related questionnaire simultaneously and collect the cervical exfoliated cells. Liquid-based cervical cytological samples of 212 women with suspicious cervical lesion were obtained before cervical biopsy at the Colposcopy outpatient clinic of Tianjin Central Hospital of Gynecology and Obstetrics(Tianjin, China), from October 2013 to April 2014., The cervical exfoliated cells used for DNA extraction. Detection of HPV types was performed with nested PCR or modified general primer PCR combining Pyrosequencing. HPV16-positive samples were further analyzed for 7 variant positions of the E6 gene with special PCR primers, then directly sequenced by pyrosequencing. Statistical analyses were carried out using the SPSS computer software(SPSS for Windows, version17.0).Results: 1. A total of 271 cases(13.6%)with HPV-positive among 2000 cases of cervical cancer screening,and the most common HPV genotype was HPV16, accounts for 107 cases(39.5%). 2. A total of 87 samples with HPV16 E6 gene-positive among HPV16 positive samples, European prototype(E-p) variant was the most common type, accounts for 107 cases(39.5%), followed by Asian(As), accounts for 10 cases(28.0%). The rest of the variation of E-G350, E-C109 and Asian American type(AA), accounts for 3 cases, 1 case and 1 case respectively.3. The percentage of HPV16 E6 variant among different age groups were different, the peak prevalence of HPV16 mutation frequency at the age ≤30, and E-p was the most common variation. However, the difference of percentages of HPV16 E6 mutation frequency in each age group was not significant(P﹥0.05). 4. HPV16 DNA infection rate was 18.6%(11/59), 51.7%(45/87), 83.9%(26/31) in 91 WNL and CINI, 90 CINII, and 31 cervical cancer groups respectively, the rate of HPV16 infection increased with the severity of cervical lesions. 5. 82 cases of HPV16 E6 gene sequencing all successful among 212 cases of in Tianjin region, two variant positions were detected, the most common type of HPV16 variants was the As, which accounts for about 54.9%(45/82), followed by E variant, accounts for about 45.1%(37/82),and the E-p was 83.8%(31/37), E-G350 was 16.2%(6/37). As variant are closely related to the severity of cervical lesions compared with the E mutation(OR: 4.337; 95% CI: 1.248-15.067; P = 0.015). 6. E-G350 variant were detected in 4 cases(9.3%) with CINII-III and 2 cases(8.3%) with cervical cancer, however inflammation and the low grade lesions group was not detected, no statistical difference between the lesion group(P >0.05). 7. The mutation rate of As variant in the younger group(≤35 years) and middle-aged group(> 35 years) was 75.9%(22/29), 50.0%(19/38) respectively, no statistical difference between between the two groups(P = 0.031).Conclusions: 1. HPV16 was the most common type among health screening women and cervical lesion patients of Tianjin. 2. E-p was the most common HPV16 E6 variation among Tianjin health screening women, however, As was the most variant among patients with high-grade cervical lesion and invasive cervical cancer, which suggests As variant was the importan risk factor for the progress of cervical lesions and cervical cancer. 3. E-G350 was the high-risk variant for high grade cervical lesions occured. 4. As variant was the risk factor for the trend of cervical lesions getting younger... |
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