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Background: Thyroid cancer is a common and frequently occurring diseases of the human head and neck, and papillary thyroid carcinoma is the most common malignancy in thyroid tumors, accounting for a large proportion. But we know very little about their development of molecular mechanisms. Biological behavior of tumor development is more than a factor of, so explore different genes expressed in papillary thyroid carcinoma, and help us to better understand the mechanism of its occurrence and guide clinical treatment. ARHI YU differential display PCR method. A new candidate tumor suppressor gene cloned from human ovarian and breast epithelial cells and cancer cells. YU study found that ARHI cells can not through the G1 phase, to prevent excessive cell proliferation inhibition of G1 cyclin-CDK activity, a large number of studies have shown that, compared with normal breast epithelial tissue and ovarian epithelial tissue, ARHI in breast and ovarian cancer, missing or low expression of ARHI is a tumor suppressor gene. Lu Z, in ovarian cancer cell line SKOV3, EMSA (electrophoretic mobility shift assay) method, E2F1 binding sites found the ARHI promoter region (-385 to -409), and the cancer cells of E2F binding activity, negative regulator ARHI E2F1 promoter activity. Nishimoto and others through the yeast two-hybrid assay and immune coprecipitation method showed that STAT3 and ARHI interactions form a complex, blocked the IL-6-mediated STAT3 into the nucleus, thereby reducing the start of STAT3 DNA binding and STAT3-dependent promoter activity. Objective: In this study, the expression of ARHI, E2F1, STAT3 protein levels in pathological morphology and immunohistochemical study of papillary thyroid carcinoma. By RT-PCR method, the expression of papillary thyroid carcinoma the ARHI gene mRNA level. In order to explore the ARHI of E2F1 expression of STAT3 in papillary thyroid carcinoma variation and possible regulatory mechanisms, and whether they have, and preliminary clarify three protein expression in papillary thyroid carcinoma occurs may role. Method: a study Select Second Hospital of Hebei Medical University, Department of Pathology, the period from December 2000 to January 2010 archive paraffin blocks, in all cases the diagnosis is clear, complete clinical data, including 88 cases of papillary thyroid carcinoma, 22 cases of thyroid gland tumor, 15 cases of thyroid follicular carcinoma, 12 cases of medullary thyroid carcinoma, 8 cases of thyroid undifferentiated carcinoma, selected 20 cases of normal thyroid tissue, immunohistochemistry detected for each indicator protein. Collect the Fourth Hospital of Hebei Medical University, March 2009 -2009 in August thyroid tumors resected specimens of 15 patients, complete clinical data, surgical resection and confirmed by pathology, including 11 cases of papillary thyroid carcinoma, medullary carcinoma 2 cases of nodular goiter cases of Hashimoto's thyroiditis 1 cases. Because medullary carcinoma, nodular goiter, Hashimoto's thyroiditis, the number of cases is too small, so only selected 11 patients with papillary thyroid carcinoma to study, In another 10 cases of normal thyroid tissue as a control for the detection of the ARHI mRNA Expression. The fresh tissue preservation at -80 ℃ spare. 2 immunohistochemical methods using immunohistochemical SP two-step method, research ARHI, E2F1, STAT3 protein expression. Specific operations in accordance with the immunohistochemistry kit. Hematoxylin graded ethanol dehydration, xylene, and mounted with neutral gum, light microscopy. 3 reverse transcription-PCR (RT-PCR) to detect mRNA expression 3.1 total cellular RNA extraction, identification and quantification of the guanidine isothiocyanate step extraction of total cellular RNA. 1% agarose gel electrophoresis integrity. 3.2 RT-PCR method anti-transcribed cDNA PCR reaction system, 94 ° C denaturation 5min, 94 ° C 50S, 62 ° C 3 0s, 30 seconds at 72 ° C, 30 cycles of amplification, PCR products on a 1.5% agarose gel electrophoresis (90V, 35min). Ultraviolet analyzer observed and photographed. 4 statistical analysis of all the data using SPSS17.0 software for statistical, X2 test, Pearson correlation analysis. Result: ARHI protein expression in papillary thyroid carcinoma in 88 cases of thyroid papillary adenocarcinoma ARHI protein positive expression rate was 65.91% (58/88), 15 cases of thyroid follicular carcinoma ARHI protein positive expression rate 66.67% (10/15), the statistical results showed that ARHI protein expression in papillary thyroid carcinoma and follicular carcinoma was significantly higher than the normal thyroid tissue (P lt; 0.05) In addition, papillary thyroid carcinoma ARHI protein expression was significantly higher than the thyroid adenoma, medullary thyroid cancer and anaplastic thyroid cancer (P lt; 0.05). Between papillary thyroid carcinoma and follicular carcinoma, thyroid adenoma, medullary carcinoma, undifferentiated carcinoma and normal thyroid tissue between ARHI protein positive expression rate differences had no statistical significance (P gt; 0.05). 2 E2F1 protein expression in papillary thyroid carcinoma in papillary thyroid carcinoma E2F1 protein positive expression rate was 70.45% (62/88); normal thyroid tissue, thyroid adenoma, thyroid follicular cancer, thyroid medullary The positive expression rates of cancer, anaplastic thyroid cancer in E2F1 were 0% (0/20), 31.82% (7/22), 80% (12/15), 83.33% (10/12), 50% (4 / 8). The chi-square test, papillary thyroid carcinoma E2F1 protein positive expression rate was significantly higher than that of normal thyroid tissue and thyroid adenoma (P lt; 0.05). Thyroid adenoma, follicular carcinoma, medullary carcinoma, and undifferentiated carcinoma E2F1 protein positive expression rate was significantly higher than that of normal thyroid (0%), the difference was statistically significant (are P lt; 0.05). Papillary thyroid carcinoma and follicular thyroid carcinoma, medullary carcinoma, and undifferentiated carcinoma between E2F1 protein expression rate difference was not statistically significant (P gt; 0.05). 3 STAT3 protein expression in papillary thyroid carcinoma thyroid papillary adenocarcinoma of STAT3 protein positive expression rate was 79.55% (70/88). Normal thyroid tissue, thyroid adenoma, thyroid follicular cancer, medullary thyroid cancer and thyroid undifferentiated carcinoma of STAT3 positive expression rate of 35% (7/22), 72.73% (16/22), 86.67% ( 13/15), 100% (12/12), 87.5% (7/8). The statistics showed that, papillary thyroid carcinoma STAT3 protein expression was significantly higher than the normal thyroid tissue (P lt; 0.05). STAT3 protein expression between papillary thyroid carcinoma and thyroid adenoma, follicular carcinoma, gland medullary carcinoma and undifferentiated carcinoma difference without statistical significance (P gt; 0.05). Follicular thyroid cancer, medullary thyroid cancer, thyroid undifferentiated cancer of STAT3 protein expression was significantly higher than in normal thyroid tissue (P lt; 0.05). 4 ARHI, E2F1, STAT3 expression in different cases related study of the situation in papillary thyroid carcinoma, correlation analysis suggest that ARHI protein expression of E2F1 protein expression was positive correlation (r = 0.238, P = 0.026). , ARHI, E2F1, STAT3 is no relationship between the normal thyroid tissue, thyroid adenoma, thyroid follicular cancer, medullary thyroid cancer and anaplastic thyroid cancer. 5 ARHI, E2F1, clinical pathological significance of STAT3 protein expression in papillary thyroid carcinoma in the statistical analysis found that In the present study, ARHI of E2F1 and STAT3 protein expression in papillary thyroid cancer age, gender, and lymph nodes metastasis correlation. 6 ARHI gene mRNA expression in papillary thyroid carcinoma and normal thyroid tissue RT-PCR results, 10 cases of normal thyroid tissue and 11 cases of papillary thyroid carcinoma in seven cases and eight cases, respectively is detected ARHImRNA expression The positive expression rate of 70%, 72.73%. Χ2 test, normal thyroid tissue and thyroid papillary adenocarcinoma of ARHI gene mRNA positive expression rate difference was not statistically significant (P gt; 0.05). Conclusion: 1 imprinted tumor suppressor gene ARHI protein low expression in normal thyroid tissue, while high expression in papillary thyroid carcinoma and follicular carcinoma, medullary thyroid carcinoma and undifferentiated carcinoma with low expression, but in papillary thyroid gland mRNA levels in cancer no significant changes, suggesting that the the ARHI role there may be differences in the organization of high protein expression may play a role in the promotion of papillary thyroid carcinoma and follicular carcinoma occurs. ARHI different, different types of thyroid cancer in E2F1 and STAT3 protein expression levels are significantly higher than the normal thyroid tissue, suggesting that E2F1 and STAT3 protein expression to promote the occurrence of different types of thyroid cancer. 3 in papillary thyroid carcinoma of E2F1 and ARHI between a positive relationship, E2F1 positively regulate expression of ARHI protein. 4 ARHI, E2F1 and STAT3 protein expression with age of onset papillary thyroid carcinoma, gender, and lymph node metastasis correlation.
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