| Objective: To investigate the genotyping characteristics of E6 295T/G and E7 647A/G polymorphic loci of HPV16 virus in different cervical tissues,to study the influence on immune-related molecules,and to explore the mechanism of these two polymorphic loci affecting immune-related molecules,and to analyze the relationship between these two polymorphic loci and the occurrence and development of cervical cancer.Methods:(1)461 cases of paraffin-embedded tissues were collected.(2)DNA was extracted from paraffin-embedded tissues,and the type of HPV infection was detected by PCR technology,and the relationship between different genotypes of infection and the degree of disease was analyzed by combining with medical records.(3)The E6 and E7 genes of 295 cervical tissue samples infected with HPV16 virus were amplified by PCR,and the distribution of HPV16 virus E6-T295 G and E7-A647 G mutations in cervical tissue was analyzed by Sanger method.(4)Immunohistochemical techniques were used to detect the distribution of IFNκ,STAT1,IRF9,IFI27,IFI44 L,OAS1,SUV39H1,STING,c GAS,RIG-I,IFNβ and IFNλ1 molecules in different cervical tissues and their expressions in different variant groups and normal groups.Results:(1)The genotypes of HPV16,HPV18,HPV52,and HPV53 in paraffin-embedded tissue samples of the cervix were detected by PCR.The infection rates of HPV16,HPV18,HPV52,and HPV53 were 64.0%,2.6%,4.8%,and 3.4% in women in Xinjiang.The total infection rate of HPV16 in Uygur ethnic group was 75.7%,the infection rate of CIN1 and CIN2 was 60.6%,the infection rate of CIN3 and cervical cancer was 82.9%.The total infection rate of HPV16 in Han ethnic group was 54.5%.The infection rate of CIN1 and CIN2 was 34.0%,the infection rate of CIN3 and cervical cancer was 66.5%.(2)The variation rate of E6-295T/G in Uyghur CIN1 patients was 41.7%.The variation rate of E6-295T/G in Uyghur patients with squamous cell carcinoma was 43.4%.The variation rate of E6-295T/G was 33.3% in Han ethnic group of CIN1 patients,and 16.1% in squamous cell carcinoma patients.The variation rate of E7-647A/G in Uygur CIN1 patients was 41.7%,and that of E7-647A/G in squamous cell carcinoma patients was 9.2%.The variation rate of E7-647A/G was 45.8% in Han ethnic group of CIN1 patients and 50.0% in squamous cell carcinoma patients.(3)The results showed that the expression levels of IFN κ,IRF9,OAS1,IFI27 and IFI44 L in normal cervical tissues without HPV infection were higher than those of CIN1,The difference was statistically significant(P<0.05).The expressions of IFN-κ,STAT1,IRF9,OAS1,IFI27 and IFI44 L in CIN3,moderately differentiated and poorly differentiated squamous cell carcinoma were higher than those of CIN1,and those in moderately differentiated cervical squamous cell carcinoma and adenocarcini were higher than those in normal tissue,and the expressions in moderately differentiated cervical squamous cell carcinoma and adenocarcini were higher,The difference was statistically significant(P<0.05).The expression levels of STING,c GAS,RIG-I,IFNβ and IFNλ1 in SUV39H1 pathway in normal cervical tissues without HPV infection were higher than those of CIN1,The difference was statistically significant(P<0.05).The expression levels of STING,c GAS,RIG-I,IFNβ and IFNλ1 in CIN3,moderately differentiated and poorly differentiated squamous cell carcinoma and adenocarcinoma were higher than those of CIN1,and those in moderately differentiated squamous cell carcinoma were higher than those in normal tissue,The difference was statistically significant(P<0.05).(4)IFN κ pathway include IFN κ,IRF9,STAT1,OAS1,IFI27 and IFI44L The expression level of IFI44 L in E6-295G/E7-647 A variant was higher than that in E6-295T/E7-647 A unvariant.The expression levels of IFN-κ,OAS1,IFI27 and IFI44 L in E6-295T/E7-647 G variant were higher than those in E6-295T/E7-647 A unvariant.The expression levels of IRF9 and OAS1 in E6-295G/E7-647 A variants were higher than those in E6-295T/E7-647 G variants,and the difference was statistically significant(P<0.05).(5)SUV39H1 signaling pathway include SUV39H1,STING,c GAS,RIG-I,IFNβ and IFNλ1.SUV39H1,RIG-1,IFNβ and IFNλ1 in E6-295G/E7-647 A variant was higher than that in E6-295T/E7-647 A prototype,The difference was statistically significant(P<0.05).There was no significant difference between E6-295T/E7-647 G and E6-295T/E7-647 A,and there was no significant difference between E6-295G/E7-647 A variant and E6-295T/E7-647 G variant.Conclusions:(1)The rate of E6-295T/G variation in cervical cancer patients infected with HPV16 in the Uygur ethnic group was higher than that in the Han ethnic group,while the rate of E7-647A/G variation in cervical cancer patients infected with HPV16 in the Han ethnic group was higher than that in the Uygur ethnic group.(2)HPV16 virus infected CIN1 tissue of immune molecules expression in IFNκand SUV39H1 of signal pathways are lower than those in normal cervical tissues without HPV16 virus infection,and the expression of these immune molecules in HPV16 infected cervical cancer tissues is higher than that in CIN1 tissues.(3)E6-295T/G variant affects the expression of both IFN κ and SUV39H1 signaling pathways,while E7-647A/G affects the expression of IFN κ signaling pathway. |
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