The Pathological Study on Expression of CK-19、VEGF-C、COX-2 and Vimentin in Grave’s Disease and Hashimoto’s Thyroiditis and the Detection of ER and PR mRNA in Hashimoto’s Thyroiditis
|School||Tianjin Medical University|
|Course||Pathology and Pathophysiology|
|Keywords||Graves' disease Hashimoto's thyroiditis CK-19 Vimentin VEGF-C COX-2 Estrogen Receptor Progesterone receptor|
Objective and Methods: Graves disease (Grave's disease, GD) and Hashimoto's thyroiditis (Hashimoto's Thyroiditis, HT) for organ-specific autoimmune thyroid disease (AITD) two main manifestations, the pathological type interstitial lymphatic related to the degree of cell infiltration. Stromal lymphoid tissue infiltration degree the GD is divided into three types, the degree of infiltration: I type (lt; 10%), II-type (10% -20%), Ⅲ type (20% -40%); Woolner's taxonomy HT is divided into three types, the degree of infiltration of the lymphoid tissue: lymphocytes (Lymphoid type, L type: 20% -40%), eosinophilic epithelial cell type (Oxyphilic epithelium type, 0 type: 40% -80%), significant the epithelial damage type (Pronounced epithelium destruction type, P: 80% -100%). CK-19, VEGF-C, COX-2 and Vimentin expression in papillary thyroid carcinoma and follicular carcinoma in the study reported more at home and abroad, less benign lesions AITD. The first part of this paper using immunohistochemistry GD and HT in the CK-19, VEGF-C, the expression of COX-2 and Vimentin detection, by analyzing four parameters in GD HT expression to explore the four indicators expression with GD and HT diagnosis, differential diagnosis, lymphocytic infiltration, the relationship between the progress of the disease. CK19, VEGF-C, COX-2 and Vimentin GD, HT-positive expression rate was used to compare the the χ2 experience (Fisher test), the positive intensity rank sum test was used to compare. Two sets of hierarchical data using bivariate correlation analysis (Bivariate) rank correlation coefficient (Spearman correlation coefficient) non-parametric test, GD and HT age two independent samples t-test was used to compare. The second part of this application reverse transcriptase PCR (RT-PCR) and fluorescence quantitative PCR (SYBR Green QRT-PCR) methods were qualitative and quantitative detection of HT, papillary thyroid carcinoma and nodular goiter of estrogen receptor ( Estrogen receptor; ER) and progesterone receptor (Progesterone receptor, PR) gene expression, ER and PR and HT correlation analysis, to explore the relationship between sex hormones and HT morbidity and illness severity the HT clinical diagnosis and treatment to provide a theoretical basis. Fluorescent quantitative detection of ER results using quantitative data completely randomized design analysis of variance. Results: 1.GD, HT sex ratio (male / female) were 1:5.33,1:18.33 GD sex ratio and sex ratio of HT by χ2 test differences were significant (χ2 = 27.32, P lt; 0.05). GD, HT average age at the time of diagnosis were 31.72,45.67 (years), t-test differences were significant (t = -7.03, P lt; 0.01), GD male average age at the time of diagnosis (40.22 years) is higher than the female average age at the time of diagnosis ( 30.13 years), t-test difference was statistically significant (t = 2.774, P lt; 0.01) The the HT men average age at the time of diagnosis (51.67 years) is higher than the female average age (45.35 years) diagnosed no statistically by t-test differences significance. 2.CK-19 is mainly expressed in the cytoplasm of epithelial cells of thyroid follicular lesions. HT in the CK-19-positive expression rate (86.2%) significantly with higher than GD (43.9%), GD Ⅲ type (81.3%) significantly with higher than GD type Ⅰ (15.8%), GD Ⅲ type significantly with higher than GD Ⅱ type (40.9%), HT-P-type (100%) was significantly higher than the HT-L (66.7%), the above differences were statistically significant (P lt; 0.05). HT CK-19-positive expression intensity than GD, the difference was statistically significant (P lt; 0.05). 3.VEGF-C positive material is mainly expressed in the cytoplasm of thyroid follicular epithelial lesion. The HT of VEGF-C of positive expression rate (96.6%) was significantly higher than the GD (56.1%) GD Ⅲ type positive rate (87.5%) significantly significantly higher than the GD Ⅰ type (36.84%), GD Ⅲ type significantly with higher than GD Ⅱ type (50.0% ), the above differences were statistically significant (P lt; 0.05). HT in the positive expression of VEGF-C was significantly higher than GD, the difference was statistically significant (P lt; 0.05). Can be seen the 4.GD thyroid follicular cells of HT-COX-2-positive immune response, distributed in the cytoplasm. GD COX-2 positive expression rate (91.2%) and HT (93.1%) are similar, but the HT positive expression was significantly higher than GD, the difference was statistically significant (P lt; 0.05). GD-positive rate for GD Ⅰ 89.5% 93.8% 90.9% GD Ⅲ type GD Ⅱ type, similar positive rate. HT-positive rate for the HT-L 85.7% and 94.7% in the HT-0, HT-P-type 100.0% positive rate is also similar, HT-O-positive expression intensity was significantly higher than the HT-L , the difference was statistically significant (P lt; 0.05). 5. Vimentin positive expression was mainly seen in the interstitial between GD and HT follicular epithelial cells and follicular epithelial cells. GD and HT Vimentin positive expression rates were 100% positive expression of Vimentin in the GD slightly higher than the HT, the difference was not statistically significant (P lt; 0.05). 6.GD in 43.9% of cases, CK-19 and Vimentin expression consistent (both positive or both negative), the Spearman rank correlation analysis, the correlation coefficient r = 0.223 (P gt; 0.05). HT in 86.2% of cases, CK-19 and Vimenitn expression consistent after Spearman rank correlation analysis, the correlation coefficient r = 0.113 (P gt; 0.05). GD in 61.4% of cases of VEGF-C and COX-2 expression, Spearman rank correlation analysis, the correlation coefficient r = 0.209 (P gt; 0.05). HT 93.1% of cases of VEGF-C and COX-2 expression, Spearman rank correlation analysis, the positive correlation between the expression of both the HT correlation coefficient r = 0.552 (P lt; 0.05). 7.RT-PCR results showed that HT, papillary thyroid carcinoma and nodular goiter visible ER-positive target bands were no the PR positive purpose bands between 100-250bp. HT in ER-positive rate was 100% (2/2), papillary thyroid cancer, ER-positive rate of 14% (1/7), nodular goiter ER-positive rate was 80% (4/5). 8 PCR results show HT in ER expression was higher than papillary thyroid cancer and nodular goiter. HT nodular goiter, HT and thyroid papillary carcinoma, papillary thyroid carcinoma with nodular goiter amount of ER expression differences were not statistically significance. Conclusion 1.GD average age at the time of diagnosis (31.72 years) than HT (45.67 years old), the difference was statistically significant (P lt; 0.05), prompts the degree of infiltration increasing as the disease progress continue to HT by GD patients with diagnosed age is gradually increased. GD male average age at the time of diagnosis (40.22 years) than females (30.13 years), and the difference was statistically significant. These results indicate that the age of onset of the different gender GD difference in the incidence of men older. GD, HT sex ratio (male / female) were 1:5.33,1:18.33, GD sex ratio and the sex ratio of HT by χ2 test differences were significant (P lt; 0.05), prompted GD and HT are female multiple HT gender difference is even greater. 2.GD and HT-lymphocyte infiltration present continuity changes that GD III type gt; GD II type gt; GD type Ⅰ HT-P gt; HT-O type gt; HT-L, CK-19 VEGF-C, COX-2 positive expression rate and positive expression of strength have the same trend, suggesting that these three indicators expression levels in ATID their lymphocyte infiltration. Vimentin in GD and HT-positive expression rate was 100%, and the positive expression of similar intensity, their lymphocytes regardless of the degree of infiltration. 3.GD in 61.4% of cases VEGF-C and COX-2 expression, but there was no correlation between the HT in 93.1% of cases VEGF-C and COX-2 expression in both a positive correlation between the expression of the HT . HT relative to GD more infiltration of lymphocytes, suggesting that the HT of COX-2 seems to be able to raise the expression of VEGF-C promote lymphocytes migrate to the thyroid tissue by blood vessels and lymphatic aggravated organization lymphocytic infiltration and destruction . Comprehensive testing of CK-19, VEGF-C, COX-2 and Vimentin GD, HT diagnosis, treatment, disease surveillance and assessment of prognosis has important clinical significance. GD, HT in the CK-19, VEGF-C, COX-2 and Vimentin indicators role in the development of the disease and the specific relationship between them needs further study. 5. RT-PCR results have a certain degree of expression of ER HT, papillary thyroid carcinoma and nodular goiter, and real-time quantitative analysis showed higher expression levels in HT, but among the three ER expression the difference was not statistically significant. ER expression in HT, papillary thyroid carcinoma and nodular goiter, estrogen through role play a biological role in the ER, estrogen is closely related to the incidence and progression of thyroid disease especially closer relationship with HT. PR is not detected in the HT, papillary thyroid carcinoma and nodular goiter three diseases expression possible reason is that the thyroid disease did not express PR.