| Objective: To clarify the distribution of different types of HPV(human papillomavirus)in Xinjiang and its relationship with the occurrence and development of cervical cancer,and analyze the viral load of HPV infection in Xinjiang.To analyze the genomic polymorphisms of HPV16 E1 and E2 of cervical cancer in xinjiang women and explore the molecular evolution law of cervical cancer caused by HPV16.Methods:(1)A total of 2042 cases of cervical exfoliated cells were collected.(2)Detection of HPV infection and determination of infection type by PCR technology,extraction of positive sample DNA and detection of HPV virus copy number.(3)A total of 165 samples of cervical exfoliated cells infected with HPV16 virus were collected for research on the evolutionary rules of HPV16 virus and analysis of sequence characteristics.The E1 and E2 genes of HPV16 virus were amplified by PCR,and the PCR amplified fragments were analyzed by Sanger dideoxy sequencing method.The evolution of HPV16 virus was analyzed by using evolutionary tree analysis software,and the evolutionary tree of HPV16 virus was established.To analyze the polymorphism characteristics of HPV16 E1 and E2 genes and clarify the evolutionary rules,grouping and source of HPV16 virus.(4)The HPV16 E2 prototype and variant expression vectors were constructed and transfected into cervical cancer He La cells and Si Ha cells.The effects of E2 prototype and variant expression vectors on HPV cancer protein E6,cell proliferation and immune-related molecules were detected.Results:(1)HPV genotypes of cervical exfoliated cells were detected by PCR reverse dot hybridization.The results showed that the infection rates of HPV16,52 and 53 in women in Xinjiang were higher,and the infection rate of high-risk HPV was higher than that of low-risk HPV and high-risk and low-risk combined infection types.Most of the infection types are HPV infection with only one genotype,followed by double infection.(2)By detecting the virus copy number of cervical samples infected with HPV16,it was found that the HPV16 virus load of cervical cancer samples was higher than that of non-cervical cancer samples.The viral load of HPV16 is higher than that of HPV52 and HPV53,and that of HPV18 is higher than that of HPV52.(3)Most of the samples infected by HPV16 are European strains,followed by Asian strains.African strain and North American/Asian strain were not found.(4)According to the analysis of HPV16 E1 gene variation,the variation of locus A978 G and locus G1473 A is significantly different between the control group and the case group.According to the analysis of HPV16 E2 gene variation,there are significant differences between the control group and the case group in the mutations of loci G1964 A,C2295A and G2385 A located in the transactivation domain of HPV16 E2,loci T2520 C,C2546T and G2585 A located in the hinge region and C2820 A,C2873T and C2923 A located in the DNA binding domain.(5)The overexpression of HPV16 E2-2585G+2820C prototype and HPV16 E2-G2585 A,E2-C2820 A,and E2-G2585A+C2820A variants could inhibit the expression of E6 protein.The overexpression of HPV16 E2 could inhibit the expression of immunemolecules IFN-κ and MHC-I.The overexpression of E2 can inhibit the proliferation of cervical cancer cells.The He La and Si Ha cells overexpressing HPV16 E2-G2585 A and E2-C2820 A variants have greater proliferation capacity than the overexpressed E2 prototype.Conclusion:(1)The infection rates of HPV16,52 and 53 among women in Xinjiang are high,and the HPV viral load of cervical cancer patients infected with HPV16 is higher than that of non-cervical cancer patients group.(2)The majority of HPV16 infection sample strains were European,followed by Asian.No African or North American/Asian strains were found.(3)Overexpression of HPV16 E2 could inhibit the expression of E6 protein,the immune molecules IFN-κ and MHC-Ⅰ.The inhibitory effect of E2 prototype on the proliferation of cervical cancer He La and Si Ha cells was greater than that of E2 mutant. |
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