Study On The Correlation Between APOBEC3s And Cervical Cancer In Uygur Women

Objective:In order to identify closely related genes in members of APOBEC3s gene famlily by analyzing HPV16 infection in Uygur cervical cancer,precancerous lesions and cervicitis,and to explore the possible molecular mechanism in the process of the members of the APOBEC3s interaction with HPV16E6 occurrence of Uygur cervical cancer.Methods:In this study,nested PCR was used to identify the cervical tissues of455 HPV positive Uygur women.The distribution of viral load in cervical cancer,precancerous lesions and cervicitis in 364 HPV16 positive Uygur women and the distribution of infection of HPV16 type and non HPV16 type Uygur in cervical lesions were analyzed.Randomly selected 120 cases of HPV16 positive cervical lesions were divided into three groups(cervical cancer group,cervical precancerous lesion group,cervicitis group),40 cases in each group.The mRNA expression of APOBEC3s in different cervical lesions was detected by Real-Time PCR method.The differentially expressed genes were screened and the protein expression in different cervical lesions was detected by Western Blotting.Finally,the screened genes were transfected into SIHA cells by lentivirus,and the gene cloning and sequencing were performed to detect the modification of HPV16E6 base.Meanwhile,the differential genes in APOBEC3s were detected,and the relationship between HPV16E6 and P53 was analyzed.To find the possible molecular mechanism of APOBEC3s in the carcinogenesis and development of cervical cancer.Results:(1)HPV16 positive cervical lesions were 364 cases and the infection rate was 80%(364/455);non HPV16 type was 91 cases and the infection rate was 20%(91/455)in 455 cases of HPV positive Uygur women cervical lesions.The infection rate of cervical cancer was 47.25%(172/364),the infection rate of cervical precancerous lesions was 40.93%(149/364),and the cervicitis infection rate was 11.81%(43/364)in 364 cases of HPV16 positive cervical lesions.The infection rate of cervical cancer was 3.30%(3/91),the infection rate of cervical precancerous lesions was30.77%(28/91)and the cervicitis infection rate was 65.93%(60/91)in non HPV16 type cervical lesions.The HPV16 viral load was positively correlated with the degree of differentiation and clinical stage of cervical cancer(P<0.05),and the correlation coefficient of Spearman was r~1=0.642,r~2=0.568.(2)There was no difference in the expression of m RNA in 120 cervical lesions(cervical cancer group,cervical precancerous lesion group,cervicitis group)about APOBEC3A,APOBEC3B,APOBEC3DE and APOBEC3H in APOBEC3 family.The expression levels of APOBEC3C,APOBEC3F and APOBEC3G mRNA were significantly different(P<0.05)in different cervical lesions.(3)The results of Western Blotting showed that the relative expression of APOBEC3C,APOBEC3F and APOBEC3G in different cervical lesions was significantly different(P<0.05).(4)The relative expression levels of HPV16E6,P53 mRNA and protein in cervical cancer cells were significantly different(P<0.05)after the overexpression of APOBEC3C,APOBEC3F and APOBEC3G.APOBEC3C,APOBEC3F and APOBEC3G genes were transfected into SIHA cells by lentivirus,and the results of gene cloning and sequencing showed that there was change in HPV16E6 gene in each group.Conclusions:(1)HPV16 infection is the main type of cervical cancer and cervical precancerous lesions with the increase of the proportion of lesions progression.With the increase of HPV16 viral load,the degree of differentiation of cervical cancer is poorer and the clinical stage is higher.(2)0Compared with the control group,mRNA gene and protein of APOBEC3C/APOBEC3F/APOBEC3G three genes were significantly changed in cervical cancer and precancerous lesions of Uygur women.These results suggest that the three genes play an important role in the occurrence and development of cervical cancer and precancerous lesions.The expression of APOBEC3A/APOBEC3B/APOBEC3DE and APOBEC3H four genes in cervical cancer and precancerous lesions of Uygur women showed no obvious change compared with cervicitis,suggesting that the possible mechanism of cervical cancer is not clear.(3)After the HPV16 positive SIHA cells in APOBEC3C/APOBEC3F/APOBEC3G cells transfected with lentiviral expression,gene cloning and sequencing indicated that HPV16E6 may have played a certain role through the base modification on cervical cancer occurrence.After overexpression of APOBEC3C/APOBEC3F/APOBEC3G,we found that the expression of HPV16E6 and P53 in cervical cancer were significantly different,suggesting that HPV16E6 may induce the increase of P53 expression and inhibit the occurrence of cervical cancer.(4)in short,the three genes of APOBEC3C/APOBEC3F/APOBEC3G may participate in the expression of HPV16E6gene in cervical cancer cells through the base modification of HPV16E6 gene and the induction of P53 expression.By regulating the expression of downstream P53 to inhibit the production of cervical cancer cells,and thus play a role in anti-virus and anti-tumor.However,the relationship between APOBEC3C/APOBEC3F/APOBEC3G and HPV16E6and the molecular mechanism leading to cervical cancer need to be further studied.