Study On The Detection Of HSP60, BRAFV600E And Immunohistochemical Package In Benign, Malignant And Borderline Thyroid Lesions

Objective at present,there are many indicators for pathological diagnosis of thyroid tumors,but it is necessary to summarize several antibodies with good sensitivity and specificity to improve the diagnostic efficiency.We selected MC,34βE12,CK19,galectin-3 and CD56 as immunohistochemical packages.As a common tumor marker,the application of HSP60 in thyroid tumors is not clear.BRAF^V600Eis a common type of gene mutation in papillary thyroid carcinoma.It is not clear whether BRAF^V600Ecan be used as an index of optimized package and the relationship between BRAF^V600Egene mutation and protein expression.The expression levels of various antibodies in benign and borderline thyroid lesions were analyzed by HSP60,BRAF^V600E,MC,34βE12,CK19,galectin-3,CD56 immunohistochemical staining（immunohistochemistry,IHC）and BRAF^V600Egene polymerase chain reaction（Polymerase Chain Reaction,PCR）.To evaluate whether each antibody can be used as a reference index for the diagnosis and differential diagnosis of benign,malignant and borderline thyroid lesions,to evaluate whether HSP60 and BRAF^V600Ecan be used as a reference index for optimizing the immunohistochemical package for thyroid papillary diagnosis,to analyze the coincidence between BRAF^V600Eimmunohistochemistry and PCR method in the diagnosis of classical subtype papillary carcinoma,and to evaluate whether the expression of BRAF^V600Eprotein can be used as a reference basis for malignant risk of PTC.Materials and methods 289 cases of thyroid papillary carcinoma（papillary thyroid carcinoma,PTC）fixed by formalin and embedded in paraffin were selected from the Department of Pathology of the first affiliated Hospital of Inner Mongolia Medical University from January 2010 to December 2018,including270cases of classical subtype（classical variant of papillary carcinoma,CVPTC）,19 cases of follicular subtype（follicular variant of papillary carcinoma,FVPTC）and 31 cases of thyroid benign lesion follicular adenoma（follicular adenoma,FA）.Tissue microarray（tissuemicroarray,TMA）.In each case,3 tissue cores were taken as tumor,benign tissue at the boundary between tumor and tumor and more than 0.5cm from tumor,and 17 cases of borderline lesions could not be identified by HE and 5 immunohistochemical packages.Immunohistochemical staining of HSP60,BRAF^V600Eprotein and 5 packages were performed in each case group,and the results were interpreted under light microscope.Paraffin samples of CVPTC17,FVPTC,borderline lesions and benign lesions were detected by PCR,and the results were analyzed by SPSS26.0 statistical software.Results（1）there was significant difference in the expression of MC,galectin-3,34βE12,CK19,CD56,HSP60 and BRAF^V600Ebetween PTC carcinoma and adjacent and distant paracancerous thyroid tissues,and between borderline lesions and adjacent and distant paratumoral thyroid tissues（P<0.05）.（2）the expression of MC,galectin-3,34βE12,CK19,CD56,HSP60,BRAF^V600Eprotein in PTC was significantly different from that in FA,（3）there was significant difference in the expression of galectin-3,CK19,HSP60 and BRAF^V600Ebetween CVPTC and FVPTC.The expressions of MC,galectin-3,34βE12,CK19,CD56,HSP60 and BRAF^V600Ein CVPTC were significantly different from those in FA,while the expressions of MC,34βE12,CK19,CD56 and HSP60 in FVPTC were significantly different from those in FA（P<0.05）.（4）the positive expressions of MC,galectin-3,34βE12,CK19,CD56 and HSP60 in FVPTC and CVPTC were significantly different from those in borderline lesions,and the positive expressions of 34βE12,CK19and CD56 in borderline lesions were significantly different from those in FA.（5）the sensitivity and specificity of MC,galectin-3,34βE12,CK19,CD56,HSP60 and BRAF^V600Eproteins in the diagnosis of PTC were 88.9%,90.1%,95.2%,89.5%,90.9%,93.3%,79.4%and 87.1%,51.6%,93.5%,74.2%,93.5%,71.0%,73.3%,respectively.The sensitivity and specificity in the diagnosis of borderline lesions were 35.3%,52.9%,41.2%,58.8%,70.6%,47.1%,23.5%and 87.1%,51.6%,93.5%,74.2%,93.5%,71.0%,73.3%,respectively.（6）the AUC of MC,galectin-3,34βE12,CK19,CD56,HSP60 and BRAF^V600Ein differential diagnosis of PTC and FA were 0.921,0.881,0.977,0.909,0.939,0.933 and 0.946,respectively,and the combined AUC was 1.（7）the coincidence coefficient（Kappa=0.821,P=0.001）between BRAF^V600Eimmunohistochemistry and PCR detection was statistically significant,indicating that the two detection methods were consistent.（8）the positive expression of BRAF^V600Ein PTC was not related to sex,age,tumor size and lymph node metastasis,but correlated with histological classification and capsule invasion.Conclusion（1）MC,galectin-3,34βE12,CK19,CD56,HSP60 and BRAF^V600Eproteins can be used as reference indexes for auxiliary diagnosis of PTC and borderline lesions,（2）galectin-3,CK19,HSP60 and BRAF^V600Eproteins have reference value in differential diagnosis of CVPTC and FVPTC,MC and 34βE12 have reference value in differential diagnosis of CVPTC,FVPTC and borderline lesions,galectin-3,CK19,CD56,HSP60 and BRAF^V600Eproteins have reference value in differential diagnosis of CVPTC and borderline lesions.（3）according to the sensitivity and specificity of each antibody in the diagnosis of PTC and borderline lesions,combined detection of various antibodies is needed to improve the diagnostic efficiency of thyroid lesions.（4）Immunohistochemistry can be used as a method to detect BRAF^V600Egene mutation in CVPTC.（5）BRAF^V600Eprotein can provide reference basis for the assessment of malignant risk of PTC.