Study On Mutation Of BRAFV600E And Tumor Infiltration Lymphocytes In Papillary Thyroid Carcinoma With Hashimoto's Thyroiditis

Objective:Thyroid carcinoma is a common endocrine tumor which includes papillary thyroid carcinoma,follicular thyroid carcinoma,Anaplastie thyroid carcinoma and Medullary thyroid carcinoma in pathology.Among them,the most common type is papillary carcinoma which accounts for 90%of all thyroid cancer.In recent years,the incidence of thyroid cancer is getting higher and higher,the United States Agency statistics show that the incidence of thyroid cancer increased at an annual rate of 6.4%.At the same time,the phenomenon of the coexistence of thyroid carcinoma with Hashimoto 's thyroiditis is more and more common,and the vast majority of pathological types is papillary carcinoma.There were many studies on the pathogenesis,prognosis and treatment of papillary thyroid carcinoma,but,few studied on the papillary thyroid carcinoma with Hashimoto 's thyroiditis.BRAF V600E mutation is the most common genetics in all papillary thyroid carcinoma.The pathogenesis of papillary thyroid carcinoma which is widely accepted is BRAFV600E mutations,which caused by Ras-Raf-MEK-ERK signaling pathway activation and stimulated cells divide indefinitely and eventually lead to tumor formation and control value.It has been reported that BRAF gene mutation can affect the clinical pathological parameters such as the external infiltration of the thyroid gland,cervical lymph node metastasis and clinical stage.However,whether the mutation of BRAFV600E is the main genetic change of Hashimoto 's thyroiditis with papillary thyroid carcinoma and its relationship with clinical pathological parameters is not clear.In clinical,pathology department mainly use the PCR method to detect BRAF gene mutation,and haven't carried out a large number of immunohistochemical detection of BRAFV600E mutation protein to aid in the diagnosis of papillary thyroid carcinoma,so the consistency of immunohistochemistry used to detect the mutation of BRAFV600E protein and PCR assay for detection of mutations in the BRAFV600E gene results need to be explored.The tissues of papillar thyroiditis carcinoma with Hahimoto 's thyroiditis and papillary thyroid carcinoma are all have lymphocytic infiltration.Whether or not the type and number of the infiltrating lymphocytes are the same,and whether it is associated with the mutation of BRAFV600E gene has not been determined.So the purpose of this paper is to explore the relationship between BRAFV600E gene mutation and papillary thyroiditis carcinoma with Hashimoto 's thyroiditis,the consistency of immunohistochemical method to detect BRAFV600E mutation protein and PCR assay for detection of BRAFV600E mutation,the situation of infiltrating lymphocytes in papillary thyroid carcinoma with Hashimoto 's thyroiditis and papillary thyroid carcinoma tissues and its relationship with BRAFV600E gene mutation.Our aim is to lay the foundation for the occurrence,development,individual treatment and prognosis study of papillary thyroid carcinoma with Hashimoto 's thyroiditis.Method:1.82 patients were selected from the Second Affiliated Hospital of Dalian Medical University who underwent surgery from January 2014 to January 2016.31 cases of thyroid papillary carcinoma,23 cases of thyroid papillary carcinoma with Hashimoto 's thyroiditis and 28 cases of Hashimoto 's thyroiditis were included in the 82 cases and their clinical and pathological data were clear.2.BRAFV600E expression in 82 cases were detected by immunohistochemistry and PCR.The lymphocytic infiltration of the 82 cases were detected by immunohistochemistry.3.SPSS 17.0 for Windows software was used for the results analyses.We use independent-samples T test,variance analysis,?2 test,Rank sum test and Spearman correlation ceefficient to analyze the data.Values of p<0.05 were concidered significant.Result:1.Clinical dataThe proportion of women of PTC+HT and HT group was higher than PTC group.Tumor diameters of PTC+HT were less than PTC group and occurred more often in the left lobe of the thyroid.All of them had statistical significance.2.BRAFV600E mutation result(1)Immunohistochemical resultPositive expression rate of BRAFV600E protein was lowest in PTC+HT group by immunohistochemical(average score is 1.65±2.23).Positive expression rate of BRAFV600E protein were 96.77%and 75.00%in PTC and HT group,and their average score were 7.61±3.35?3.14±2.23 respectively.All of them had statistical significance.(2)PCR result The mutation of the BRAFV600E gene expression was different between the three groups.The BRAF geneV600E mutation rate of PTC+HT group were significantly lower than the group of PTC.The PCR results of HT group were all wild types.(3).Immunohistochemical&PCRThere were 61 cases who expressed BRAFV600E mutation protein by immunohistochemical of all the 82 cases,but 33 cases which had BRAF gene V600E mutation by PCR.The immunohistochemical results were all positive in the cases which had PCR positive results.The intensity of positive protein were all<(++)in the cases which had different results by immunohistochemical and PCR methods.(4).Gene sequencing resultAll of the 28 cases which expressed BRAFV600E protein but had no BRAF gene V600E mutation were all wild types and consistented with PCR result.(5).To compare with PCR result,sensitivity and specificity of immunohistochemical result were 100%(33/33)and 42.86%(21/49)respectively.3.BRAF gene V600E mutation&Clinical dataThe clinical data(age,sex,t umor diametre,lymphatic metastasis,capsule invasion)had no clear relationship with the result of BRAFV600E mutation in group PTC+HT and PTC.4.Lymphocytic infiltration(1)CD3+lymphocytesAll of the three groups had different numbers of lymphocytic infiltration.Most of them were CD3+cells and few were CD20+cells.HT group had the largest number of lymphocytes,PTC group had the least number of lymphocytes and the number of lymphocytes in PTC+HT group is medium.(2)CD4+/CD8+lymphocytes rateThe lymphocytes rate of CD4+/CD8+in three groups all had statistical significance.(3)The number of CD3+lymphocytes and CD4+/CD 8+lymphocytes rate had no relationship with BRAF gene V600E mutation result in both PTC+HT and PTC group.5.To compare the capsule invasion and no capsule invasion group,the front group had larger tumor diameter,lymph,higher node metastasis rate and BRAFV600E mutation rate,and lower CD3+lymphocytes number,CD4+/CD8 lymphocytes rate.Conclusion:1.To compare with PTC patients,PTC+HT happened more often in woman and had smaller tumor diameter,lower BR AFV600E mutation.PTC+HT group had larger number of tumor infiltration T cells and higher rate of CD4+/CD8+lymphocytes rate.All the results hinted that the prognosis of PTC+HT patients may be better than PTC group.2.The rate of BRAFV600E mutation in PTC+HT group is lower than PTC group.It hinted that BRAFV600E mutation may not the major genetic change.3.To compare the capsule invasion and no capsule invasion group,the front group had larger tumor diameter,lymph,higher node metastasis rate and BRAFV600E mutation rate,and lower CD3+lymphocytes number,CD4+/CD8 lymphocytes rate.This hinted that BRAFV600E mutation,less numbers of tumor infiltration T cells and decline of CD4+/CD8 lymphocytes rate may be the factor of bad prognosis.4.Immunohistochemistry can be the early screening method to test the BRAFV600E mutation.The senstivity and specificity of PCR method are both better than immunohistochemistry.