Relationship Between HPV16 Gene Mutation And Cervical Lesions

Objectives:To investigate the relationship between HPV16 E6 and E7 gene mutations and the occurrence and development of cervical lesions Methods:Under the condition of informed consent,120 cases of HPV positive cervical exfoliated cells were collected from the second Hospital of Jilin University from August 2020 to December 2021.There were 38 cases of cervical cancer,32 cases of CIN,25 cases of cervical exfoliated cells with HPV positive but negative cytology or cervical biopsy,and the clinical data of each sample were collected.We extracted the DNA from the sample,designed the primers of HPV16 E6 and E7 genes and verified their specificity.The E6 and E7 gene fragments in the samples were amplified by PCR,and the obtained gene fragments were sequenced to detect the existence of HPV16 infection in these samples and to find the mutation sites of HPV16 E6 and E7 genes.Statistical methods were used to analyze the relationship between these variants and the occurrence and development of cervical lesions.Results:（1）Statistical analysis showed that there was significant difference in the average age between cervical cancer group and CIN and infection group（P < 0.05）,but there was no distinct difference in the average age between CIN group and infection group（P =0.246）;There was no significant difference in the number of pregnancies,times of delivery and mode of delivery among cervical cancer group,CIN group and infection group（P > 0.05）;There were 28（73.68%）cases of cervical cancer,32（56.14%）cases of CIN,and 7（28.00%）cases in the infection group were infected with HPV16.There was significant difference in the rate of HPV16 infection among cervical cancer group,CIN group and infection group（P = 0.001）.With the increase of the degree of cervical lesions,the rate of HPV16 infection increased.（2）There were 19 cases（65.52%）,19 cases（59.38%）and 4 cases（57.14%）of E6 gene mutation in cervical cancer group,CIN group and infection group respectively.The most common mutation sites in the three groups were T178G/A（D25E）、T350G（L83V）、A131C（R10R）and T241G（A46A）.A131C（R10R）and T241G（A46A）were synonymous mutations.Statistical analysis showed that there was no significant difference in the incidence of HPV16E6 gene mutation among cervical cancer,CIN and infection groups（P > 0.05）.（3）E7 gene mutation occurred in 24 cases（82.76%）,25 cases（78.13%）and 4 cases（57.14%）in cervical cancer group,CIN group and infection group respectively;The common mutation sites in the three groups were A647G（N29S）、G666A（E35E）、T843C（C94C）、T846C（S95S）and T760C（L67L）;A647G（N29S）and T846C（S95S）mutations occurred simultaneously.Statistical analysis showed that there was no significant difference in the incidence of HPV16E7 gene mutation among cervical cancer,CIN and infection groups（P > 0.05）;There was no significant difference in the distribution of A647G（N29S）、G666A（E35E）、T843C（C94C）、T846C（S95S）and T760 C among the three groups（P > 0.05）.（4）In this study,it was found that T178G/A（D25E）、A647G and T846 C mutations often occurred at the same time.The co-mutations of T178G/A-A647G-T846 C in cervical cancer,CIN and infection groups were 11 cases（37.93%）,14 cases（43.75%）and 2 cases（28.57%）.Statistical analysis showed that there was no significant difference in the distribution of T178G/A-A647G-T846 C co-mutation among the three groups（P > 0.05）.（5）Through the analysis of multiple mutation sites,only Eur and As were found this time,and Afr、AA and NA were not found.Conclusion:1.The average age of cervical cancer in this study was higher than that of CIN and HPV infection,and there was no significant difference between CIN and HPV infection.2.The times of pregnancy and delivery and mode of delivery have nothing to do with the occurrence and development of cervical lesions in this study.3.With the aggravation of cervical lesions,the rate of HPV16 infection increased;HPV16 infection can promote the progression of cervical lesions.4.HPV16 E6 gene mutation and the common mutation sites T178G/A（D25E）、T350G（L83V）、A131C（R10R）and T241G（A46A）were not related to the progression of cervical lesions.5.HPV16 E7 gene mutation and the common mutation sites A647G（N29S）、G666A（E35E）、 T843C（C94C）and T846C（S95S）were not related to the progression of cervical lesions.6.A647G（N29S）and T846C（S95S）mutations in E7 gene occur simultaneously;T178G /A-A647G-T846 C co-mutation was not associated with the progression of cervical lesions.7.In this experiment,only Eur and As were found,and no other variants were found.