| Objective: To compare the differences of clinicopathological data between Uyghur and Han women with advanced cervical squamous cell carcinoma(SCC)in Xinjiang province,to evaluate the clinical characteristics and prognostic factors according to follow-up study and survival;2)To investigate the distribution of human leukocyte antigen(HLA)-I and II alleles and haplotypes in advanced SCC and control healthy subjects,from both Uyghur and Han populations,and analysis the relationship between HLA and human papillomavirus(HPV)status and clinical outcome;3)To screen the HLA restricted HPV16 E6 and E7 antigen specific peptides from advanced SCC,and provide the target peptides for further clinical research.Methods: 1)A total of 853 advanced SCC patients were collected from the Affiliated Tumor Hospital of Xinjiang Medical University.The clinicopathological parameters were compared among the different groups and the correlation of clinical features and outcome were analyzed;2)A total of 175 advanced SCC patients(100 Uyghur and 75 Han)and 200 control subjects(100 Uyghur and 100 Han)from Xinjiang province were genotyped for HLA-I and II alleles using polymerase chain reaction sequence-based typing(PCR-SBT).The different frequencies of HLA alleles and haplotypes between patients and control subjects were compared and the correlations were analyzed between HLA distribution and HPV status and prognosis;3)No history of infectious disease and HPV16 type advanced SCC patients,and control subjects with HPV negative from the front part were selected.Specific immune response level of peripheral T cells in Peripheral blood mononuclear cells(PBMC)of patients and control subjects to entire protein antigen peptide pools of HPV genome sequences expression were measured by using ELISpot assay.Meanwhile,stimulate the SCC patients to produce IFN-? by the synthetic E6 and E7 antigen peptide.Combined with the results of the second part of HLA,the HLA restrictive HPV16 E6 andE7 specific peptides were screend among the patients.Results: 1)The Uyghur and Han patients with advanced SCC had significant difference in the menarche and marriage age,number of marriages and pregnancy,number of children and abortion,family history of malignant tumor,BMI,PNI,the appearance of cervix,PLTc,FBG,OS,DSS and so on.The ethnity,FIGO stage,SCC-Ag,FBG and NLR were the independent prognostic factors of DSS while FIGO stage,SCC-Ag,PLTc,FBG,NLR and family history of cancer were of the DFS in advanced SCC patients;2)Significantly higher frequencies of A*01:01,A*03:01,A*03:02,B*14:02,Cw*05:01,DRB1*13:01,DQB1*05:02 and DQB1*06:02,and lower frequencies of A*24:02,A*11:01,A*30:01,B*15:01,B*40:01,Cw*14:02,DRB1*09:01 and DQB1*03:03 were observed in the Uyghur control subjects compared with the Han control subjects.The frequencies of B*35:03,DRB1*07:01,DRB1*04:04,DRB1*04:07,DRB1*08:03,DRB1*15:02,DQB1*02:01 alleles and Cw*01:02-DRB1*09:01,Cw*01:02-DQB1*03:03,DRB1*07:01-DQB1*02:01,DRB1*15:01-DQB1*06:02 haplotypes in patients were significantly higher than in control subjects while the frequency of B*15:01,B*44:02,B*58:01,Cw*03:02,DRB1*04:03,DRB1*08:01,DRB1*12:01,DRB1*16:01,DQB1*06:05 alleles and B*58:01-Cw*03:02,DRB1*03:01-DQB1*02:02,DRB1*04:01-DQB1*03:02,DRB1*09:01-DQB1*03:01haplotypes in patients were lower than in control subjects.The frequency of DRB1*07:01,DRB1*15:02,DQB1*02:01 alleles and A*01:01-Cw*06:02,A*01:01-DRB1* 07:01,B*50:01-Cw*06:02,DRB1*07:01-DQB1*02:01,DRB1*15:02-DQB1*06:01,Cw*06:02-DRB1*07:01-DQB1*02:01 haplotypes in Uyghur patients were higher than Uyghur control subjects.However,the frequency of B*44:02,B*58:01,Cw*03:02,Cw*05:01,DRB1*04:01,DRB1*12:01,DQB1*05:02 alleles and DRB1*04:01-DQB1*03:02,DRB1*12:01-DQB1*03:01,DRB1*13:01-DQB1*06:02 haplotypes were lower than Uyghur control subjects.The frequency of A*02:07,B*46:01,DRB1*04:07,DRB1*08:03 alleles and A*02:07-B*46:01,A*02:07-Cw*01:02,A*02:07-Cw*03:04,B*46:01-Cw*01:02,B*46:01-DRB1*09:01,B*46:01-DQB1*03:03,Cw*01:02-DRB1*09:01,Cw*01:02-DQB1*03:03,DRB1*08:03-DQB1*06:01,DRB1*09:01-DQB1*03:03haplotypes in Han patients were higher than Han control subjects.However,the frequency of A*24:02,DRB1*04:03,DRB1*08:01,DRB1*12:01 alleles and A*24:02-Cw*04:01,DRB1*12:01-DQB1*03:01 haplotypes were lower than Han control subjects.In addition,B*15:01,B*44:02 and Cw*05:01 alleles were associated with HPV while B*44:02 ? DRB1*08:01 and DRB1*07:01 alleles were associated with HPV16 in advanced SCC patients.A*01:01-Cw*06:02,A*01:01-DRB1*07:01,B*50:01-Cw*06:02and Cw*06:02-DRB1*07:01-DQB1*02:01 haplotypes were associated with HPV while B*44:02 and DRB1*08:01 alleles and the haplotype Cw*06:02-DRB1*07:01-DQB1*02:01 were associated with HPV16 in Uyghur patients with advanced SCC.The allele A*24:02 in advanced SCC patients treated with concurrent radiochemotherapy was associated with increased disease-specific survival(DSS)while DRB1*09:01 and DQB1*02:01 alleles haplotypes were associated with reduced disease-free survival(DFS).The FIGO stage,treatment condition,NLR,PLTc,FBG and A*24:02 were the independent prognostic factors of DSS while the ethnity,FIGO stage,PLTc and DRB1*09:01 were of the DFS in advanced SCC patients;3)The frequency of HPV E6,E7(and intensity of E7)antigen peptide pools specific CTL immune reaction in SCC patients were significantly higher than healthy control subjects.The intensity of E7 antigen peptide pools specific CTL immune reaction in Uyghur patients was significantly higher than Uyghur control subjects.The frequency and intensity of E7 antigen peptide pools specific CTL immune reaction in Han patients were significantly higher than Han control subjects.Their differences were statistically significant.According to the results of second part,the allele HLA-A*24:02 and DRB1*09:01 were closely related to the onset of advanced SCC.The antigenic epitope of E6 and E7 were further screened according to the target gene.The positive reactions were shown in HPV16 E6 “Peptide-14 DLLIRCINCQKPLCP EEK” and E7 “Peptide-9 LRLCVQSTHVDIRTLEDL” restricted by the allele A*24:02,and HPV16 E6 “Peptide-15 CQKPLCPEEKQRHLDKKQ” by the allele DRB1*09:01 in advanced SCC patients with HPV16 positive.Conclusion: 1)The incidence,clinicopathological features and prognosis of the Uyghur and Han advanced SCC patients are different from each other.The ethnity,FIGO stage,SCC-Ag,PLTc,FBG and NLR are the independent factors related to the survical of the patients with advanced SCC;2)There are some differences in the distribution of HLA polymorphisms between advanced SCC patients and control subjects.The HLA alleles and haplotypes that are susceptible or protective effects on SCC also have certain impact on the identification and clearance of HPV infection and the evolution of SCC.The allele * 24:02 may play the protective role in DSS while DRB1*09:01 and DQB1*02:01 play the susceptible role in DFS.The ethnity,FIGO stage,treatment condition,NLR,PLTc,FBG,A*24:02 and DRB1*09:01are the independent factors related to the survical of the patients with advanced SCC.The alleles A*24:02 and DRB1*09:01 may be the HLA target gene loci of the SCC;3)HPV16 antigen E6,E7 proteins may be oncogenic proteins leading to the occurrence and development of SCC,potentially ideal targets for cervical cancer immunotherapy.Theallele HLA-A*24:02 restrictive HPV16 E6 “Peptide-14 DLLIRCINCQKPLCPEEK” and E7 “Peptide-9 LRLCVQSTHVDIRTLEDL”,and DRB1*09:01 restrictive HPV16 E6“Peptide-15 CQKPLCPEEKQRHLDKKQ” may provide a research direction for the development of cervical cancer vaccines. |
PDF Full Text Request