Analysis Of BRAF And RET Gene Mutations In Papillary Thyroid Carcinoma Combined With Hashimoto’s Thyroiditis

Part I Comparison between Papillary Thyroid Carcinoma and Papillary Thyroid Carcinoma Combined with Hashimoto’s ThyroiditisObjective:To investigate the characteristics and differences between papillary thyroid carcinoma（PTC）and PTC combined with Hashimoto’s thyroiditis（HT）.Methods:We retrospectively analyzed the clinical data of 5653 patients with thyroid diseases who underwent surgical treatment in the general surgery department of Tianjin Medical University General Hospital from January,2010 to December,2014.The percentages of cases with PTC and PTC combined with HT that received surgical treatment were evaluated.Moreover,the clinical characteristics of both groups were also analyzed.Results:1.Of all 5653 cases,441 HT patients（including 133 cases combined with PTC）and 1332 PTC patients underwent surgical treatment.The incidence of papillary carcinoma was significantly higher in PTC patients combined with HT than that in PTC（P=0.02）.2.There was no significant difference in age between PTC combined with HT and PTC patients.3.There was a significant difference in terms of gender between PTC combined with HT and PTC patients（P=0.002）.4.There was a significant difference in the percentage of microcarcinoma between PTC combined with HT and PTC patients（P=0.019）.5.There was a significant difference in the AJCC staging between PTC combined with HT and PTC patients（P=0.013）.Conclusions:HT patients are more likely to develop PTC than the patients without HT that underwent thyroid surgery.There was no significant difference in age between PTC combined with HT and PTC patients.In HT patients,the incidence of PTC was higher in female than that in male.PTC patients combined with HT have a better staging than PTC patients do.Part II Analysis of BRAF^V600E Mutation and RET/PTC Rearrangement in PTC and PTC Combined with HTObjective:To investigate the clinical significance of BRAF^V600E mutation and RET/PTC rearrangement in PTC and PTC combined with HT.Methods:PCR,nested PCR and gene sequencing assays were employed to examine the BRAF point mutation and RET/PTC rearrangement in cancer tissue from 93 HT combined PTC patients and 90 PTC patients.We analyzed the clinical and pathological characteristics of the BRAF^V600E mutation and RET/PTC rearrangement.Results:1.According to AJCC staging,of 93 PTC combined with HT cases,65,13,11,and 4 cases were staged as I,II,III,and IV,respectively.Of 90 PTC cases,43,10,24,and 13 cases were staged as I,II,III,and IV,respectively.There is a significant difference in terms of staging between the two groups（P=0.02）.2.There were 22and 34 cases with BRAF^V600E mutation in cancer tissue from PTC combined with HT group and PTC group,respectively.There was a significant difference in the percentage between the two groups（23.7%,22/93 vs.37.8%,34/90,P=0.038）.3.Of93 PTC combined with HT cases,65,13,11,and 4 cases were staged as I,II,III,and IV,respectively.Of 22 cases with BRAF^V600E mutation,12,4,4,and 2 cases were staged as I,II,III,and IV,respectively.There was no significant difference in terms of staging between BRAF^V600E mutation and wild type cases（P=0.224）.4.Of 90 PTC combined with HT cases,43,10,24,and 13 cases were staged as I,II,III,and IV,respectively.Of 34 cases with BRAF^V600E mutation,9,4,13,and 8 cases were staged as I,II,III,and IV,respectively.There was a significant difference in terms of staging between BRAF^V600E mutation and wild type cases（P=0.011）.5.There were 29 and16 cases with RET/PTC rearrangement in cancer tissue from PTC combined with HT group and PTC group,respectively.There was a significant difference in the percentage between the two groups（31.2%,29/93 vs.17.8%,16/90,P=0.035）.6.Of29 cases with RET/PTC rearrangement in PTC combined with HT group,22,3,3,and 1 cases were staged as I,II,III,and IV,respectively.There was no significant difference in terms of staging between RET/PTC mutation and wild type cases（P=0.858）.7.Of 16 cases with RET/PTC rearrangement in PTC group,9,4,2,and 1cases were staged as I,II,III,and IV,respectively.There was no significant difference in terms of staging between RET/PTC mutation and wild type cases（P=0.764）.Conclusions:1.The cancer staging in PTC combined with HT patients is earlier than that in PTC patients.2.BRAF^V600E mutation is more common in PTC patients than in PTC combined with HT patients.3.There is no significant correlation between BRAF^V600E mutation and cancer staging in 93 PTC combined with HT patients.4.There is a significant correlation between BRAF^V600E mutation and cancer staging in90 PTC patients.5.RET/PTC rearrangement is more common in PTC combined with HT patients than in PTC patients.6.There is no significant correlation between RET/PTC rearrangement and cancer staging in 93 PTC combined with HT patients.7.There is no significant correlation between RET/PTC rearrangement and cancer staging in 90 PTC patients.Part III Relationship between HLA Subtypes and Thyroid Disease and BRAF MutationObjective:To investigate the relationship between HLA subtypes and PTC,HT,PTC combined with HT and nodular goiter patients,and analysis BRAF^V600E mutations in the immune antigen recognition and presentation of the role.Methods:PCR-SSP method was used in 164 cases of thyroid patients for HLA subtype testing,statistical frequency of their HLA subtypes was analysised,and compared with the date provied by Chinese Marrow Donor Program.Bioinformatics software SYFPEITHI was used to assess the role of BRAF^V600E mutations in immune recognition.Results:（1）HLA-A subtypes frequency in the normal population and PTC patients was significantly different.（2）HLA-A subtypes frequency in PTC patients combined with HT and normal population was significantly different.（3）HLA-A subtypes frequency in different thyroid disease was significantly different.（4）HLA-DR subtypes in the normal population and PTC patients wasn’t significantly different in the frequency distribution.（5）Wild-type BRAF and mutation had different binding score with the HLA-A2,HLA-A24 subtype.Conclusion:（1）The frequency distribution of HLA subtype had certain relevance with PTC and PTC combined with HT.（2）Wild-type BRAF and mutantion had different binding score with the HLA-A2,HLA-A24 subtype,which might be a factor of the occurance of PTC.