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

Objectives and methods:Graves disease (GD) and hashimoto's thyroiditis (HT) are the two main forms of organ specific autoimmune thyroid diseases (AITD). The histotype of GD and HT was related to infiltrating lymphocytes in thyroid. According to the degree of interstitial infiltrating lymphocytes, GD will be divided into three types. They are GDⅠtype(<10%), GDⅡtype(10%～20%), GDⅢtype(20%～40%); According to the Woolner' s classification, HT was divided into three types:HT-L(20%～40%), HT-O(40%～80%), HT-P(80%～100%). There are many Home and abroad literatures about the study of CK-19, VEGF-C, COX-2 and Vimentin in PTC and FTC, However the literatures about autoimmune thyroid disease are fewever. We detected the four indicators expression in GD and HT with the immunohistochemical method. By analyzing the four indicators expression, we find the relationship with GD and HT of the diagnosis, differential diagnosis, lymphocytic infiltration and the progress of the disease.χ2 test(Fisher test)is used to compare the positive rate of CK19,VEGF-C,COX-2 and Vimentin in GD and HT. Wilcoxon test was used to compare the intensity of the four indexes expression. Spearman correlation coefficient non-parametric test is used to rank the relevance of two groups of grade material, two independent samples t-test was used to compare the age of GD and HT。We used RT-PCR and SYBR Green QRT-PCR in part two to ascertain the relationship of estrogen receptor(ER) and progesterone receptor(PR) with Hashimoto's thyroiditis in the pathogenesis, diagnosis, treatment and clinical diagnosis, providing a theoretical basis. ER results use the completely randomized design analysis of variance under the quantitative data.Results:1. The gender (male/female) ratios of GD and HT were 1:5.33 and 1:18.33 respectively, GD and HT sex ratio showed significant difference (χ2=27.32, P<0.05). GD,HT average diagnosed aged 31.72,45.67(years) showed significant difference(t=-7.03, P<0.05). GD diagnosed male average age(40.22 years) was older than in females diagnosis average age(30.13 years) (t=2.774, P<0.05). HT diagnosed male average age(51.67 years) was older than in females diagnosed average age(45.35 years), but the difference was not significant.2. CK-19 was mainly expressed in the lesions of the cytoplasm of thyroid follicular epithelial cells. CK-19 positive rate(86.2%) in HT was bigger than GD(43.9%), GDⅢtype(81.3%) was bigger than GDⅠtype(15.8%), GDⅢtype was bigger than GDⅡ(40.9%), HT-P type(100.0%) was bigger than HT-L type(66.7%), all these differences were statistically significant(P<0.05). CK-19 expression intensity in HT was higher than GD with Statistically significant (P<0.05).3. VEGF-C was mainly expressed in the lesions of the cytoplasm of thyroid follicular epithelial cells. VEGF-C positive rate(96.6%) in HT was bigger than GD(56.1%), GDⅢtype(87.5%) was bigger than GDⅠtype(36.8%), GDⅢtype was bigger than GDⅡ(50.0%), these differences were statistically significant(P<0.05). VEGF-C expression intensity in HT was higher than GD with Statistically significant (P<0.05).4. COX-2 was mainly expressed in the lesions of the cytoplasm of thyroid follicular epithelial cells from GD and HT. COX-2 positive rate(96.6%) in GD was close to HT(93.1%), but COX-2 expression intensity in HT was higher than GD with Statistically significant (P<0.05). The positive rate of all types of GD were close, they were that GD I type was 89.5%, GDⅡtype was 90.9%, GDⅢtype was 93.8%. The positive rate of all types of HT were also close, they were that HT-L type was 85.7%, HT-O type was 94.7%, HT-P type was 100.0%. COX-2 expression intensity in HT-O type was higher than HT-L type with Statistically significant (P<0.05).5. Vimentin was expressed both in the mesenchyma and in some of the cytoplasm of thyroid follicular epithelial cells from GD and HT. All Vimentin positive rates in GD and HT were 100.0%, Vimentin expression intensity in GD was higher than HT, but it has no Statistically significant (P>0.05).6.86.2% of HT of the expression of CK-19 and Vimentin were consistent (both positive or both negative), through the spearman rank correlation analysis, r=0.113(P>0.05). Only 43.9% of GD of the expression of CK-19 and Vimentin were consistent, through the spearman rank correlation analysis, r=0.223 (P>0.05).61.4% of GD of the expression of VEGF-C and COX-2 were consistent, through the spearman rank correlation analysis, r=0.209 (P>0.05).93.1% of HT of the expression of VEGF-C and COX-2 were consistent, through the spearman rank correlation analysis, they were positively correlated, r=0.552(P<0.05).7. RT-PCR results showed that HT,papillary thyroid carcinoma(PTC) and nodular goiter can be seen in ER-positive bands in the 100～250 bp region, PR positive bands were not seen. ER positive rate was 100%(2/2) in HT,14%(1/7) in PTC and 80%(4/5)in nodular goiter respectively.8. Real-time quantitative PCR results showed that the expression of ER in HT was higher than PTC and nodular goiter. There was no difference in the expression of ER between HT and nodular goiter, HT and PTC, PTC and nodular goiter.conclusion1. GD diagnosed average age(31.72 years) was lower than HT diagnosed average age(45.67years) with Statistically significant (P<0.05). Suggesting that the diagnosed age of patients increased continuously with the progression of the disease. GD diagnosed male average age(40.22 years) was older than female (30.13 years) with Statistically significant (P<0.05), Suggesting that the age of onset of GD in male was higher than the female. GD,HT gender (male/female) ratios were 1:5.33,1:18.33, GD and HT sex ratio showed significant difference,χ2=27.32(P<0.05). Suggesting that GD and HT were more common in women.2. The degree of lymphocyte infiltration in various types of GD and HT showed continuous changes. They are that GDⅢtype>GDⅡtype >GD I type, HT-P type >HT-O type >HT-L type. The positive rates and expression intensity of CK-19,VEGF-C,COX-2 showed the same trend. Suggesting that the expression of the three indicators was related to the degree of infiltrating lymphocytes in autoimmune thyroid disease.3.61.4% of GD of the expression of VEGF-C and COX-2 were consistent, but there is no correlation between the two indicators.93.1% of HT of the expression of VEGF-C and COX-2 were consistent, and there is positive correlation between the two indicators. Compared with GD, the degree of lymphocytic infiltration was higher in HT. Suggesting that COX-2 seems to up-regulated VEGF-C to promote lymphocyte migration from the blood and lymph vessels to the thyroid and Increased lymphocyte infiltration and destruction of the organization.4. The aggregate analysis of CK-19, Vimentin, VEGF-C and COX-2 was very important to clinic for diagnosing, treating, disease surveillance and evaluating prognosis of GD and HT. Their function in GD and HT and the relationship of them need further research.5. RT-PCR results showed the expression of ER in HT,PTC and nodular goiter, Real-time quantitative PCR results showed the quantity of ER expression in HT was higher than PTC and nodular goiter, but there was no Statistically significant. Among the three diseases, the difference also has no Statistically significant. Estrogen played biological effects through ER, So Estrogen had close relationship with the three disease in Pathological mechanism, especially with HT. We did not detected expression of PR in HT,PTC and nodular goiter, It may be that they didn't express PR.