Adenocarcinoma, Follicular ; genetics ; metabolism ; pathology ; Biomarkers, Tumor ; genetics ; Diagnosis, Differential ; Humans ; Matrix Metalloproteinase 1 ; biosynthesis ; genetics ; Telomerase ; biosynthesis ; genetics ; Thyroid Neoplasms ; genetics ; metabolism ; pathology ; ras Proteins ; biosynthesis ; genetics

OBJECTIVETo study Twist expression in thyroid papillary carcinoma (PTC) by immunohistochemistry and to assess its usefulness as marker in the differential diagnosis of PTC, follicular adenomas (FA) and benign papillary lesions (BPL).

METHODSFifty cases of PTC, 48 cases of FA and 47 cases of BPL were evaluated using manual tissue chip and SP immunohistochemical stain to detect the expression of Twist and HBME-1, and comparing the staining to that of cytokeratin 19 (CK19).

RESULTSIn PTC, positive rates of Twist, HBME-1 and CK19 were 100% (48/48), 94.0% (47/50) and 78.0% (39/ 50) respectively; in FA, positive rates were 0, 6.7% (3/45) and 0 respectively; in BPL, positive rates were 7.0% (3/34), 2.1% (1/47) and 0, respectively. The differences between PTC and FA and between PTC and BPL were both statistically significant (P = 0. 000). The sensitivity of Twist, HBME-1 and CK19 was 100%, 94.0% and 78.0%; the specifity was 96.4%, 95.7% and 100%; overall accurary was 97.7%, 95.1% and 91.9%, respectively.

CONCLUSIONSPositive rates of Twist is higher than the other markers in PTC. Immunohistochemical staining of Twist has important significance in the differential diagnosis of thyroid lesions. Twist immunohistochemistry maybe helpful in diagnosis and differential diagnosis of PTC.

Adenocarcinoma, Follicular ; metabolism ; Adenocarcinoma, Papillary ; pathology ; Adenoma ; diagnosis ; metabolism ; Biomarkers, Tumor ; immunology ; Carcinoma, Papillary ; diagnosis ; metabolism ; pathology ; Carcinoma, Papillary, Follicular ; metabolism ; Diagnosis, Differential ; Galectin 3 ; genetics ; metabolism ; Immunohistochemistry ; Keratin-19 ; genetics ; Keratins ; genetics ; metabolism ; Nuclear Proteins ; genetics ; metabolism ; Thyroid Neoplasms ; diagnosis ; metabolism ; pathology ; Thyroid Nodule ; pathology ; Twist-Related Protein 1 ; genetics ; metabolism

Adenocarcinoma, Follicular ; genetics ; metabolism ; pathology ; Carcinoma, Papillary ; genetics ; metabolism ; pathology ; Carcinoma, Renal Cell ; genetics ; metabolism ; pathology ; Diagnosis, Differential ; Humans ; Immunohistochemistry ; Kidney Diseases, Cystic ; genetics ; metabolism ; pathology ; Kidney Neoplasms ; genetics ; metabolism ; pathology ; Thyroid Neoplasms ; genetics ; metabolism ; pathology ; Translocation, Genetic

Adenocarcinoma, Follicular ; epidemiology ; genetics ; metabolism ; pathology ; Carcinoma, Papillary ; pathology ; DNA-Binding Proteins ; metabolism ; Diagnosis, Differential ; Genes, ras ; genetics ; Humans ; Point Mutation ; Prognosis ; Thyroglobulin ; metabolism ; Thyroid Neoplasms ; epidemiology ; genetics ; metabolism ; pathology ; Transcription Factors

OBJECTIVETo evaluate the expression of BRAF V600E mutation in 240 Chinese patients with thyroid lesions.

METHODSTwo hundred and forty Chinese patients with thyroid lesions, including 129 papillary thyroid carcinomas (PTC), 12 follicular carcinomas, 4 medullary carcinomas, 30 adenomas, 30 nodular goiters, and 35 papillary hyperplasia. DNA was extracted from thyroid biopsy and paraffin embedded thyroid tissues, and the expression of BRAF V600E mutation was detected by polymerase chain reaction and DNA sequencing assays.

RESULTSThe presence of BRAF V600E mutation was found in 61 of the total group of 240 cases (25.4%). It was only detected in PTC (47.3%), and not detected in other types of malignant and benign thyroid lesions. There was a statistically significant difference between the expression of BRAF V600E mutation in classic type PTC (49.6%) and in follicular type PTC (12.5%,P < 0.05), but statistical data did not show any correlation between BRAF V600E mutation and clinicopathologic parameters in PTC (P > 0.05).

CONCLUSIONSBRAF V600E mutation has a significant correlation with PTC and the detection of BRAF V600E mutation may be used as an important prognostic marker of PTC. Our new method of DNA extraction from paraffin embedded tissues is efficient and inexpensive.

Adenocarcinoma, Follicular ; genetics ; metabolism ; Adenoma ; genetics ; metabolism ; Adult ; Biomarkers, Tumor ; genetics ; Carcinoma, Papillary ; genetics ; metabolism ; Codon ; DNA Mutational Analysis ; DNA, Neoplasm ; genetics ; isolation & purification ; Female ; Goiter, Nodular ; genetics ; metabolism ; Humans ; Male ; Middle Aged ; Point Mutation ; Proto-Oncogene Proteins B-raf ; genetics ; metabolism ; Thyroid Neoplasms ; genetics ; metabolism

OBJECTIVETo investigate the expression of motilin and its precursor mRNA in normal human thyroid. To compare the expression differences of motilin and it precursor mRNA between normal thyroid and intestines. To study the expression of motilin and its precursor mRNA in human thyroid tumors and their clinical implications.

METHODSRT-PCR, Southern blot and molecular cloning were used to detect motilin transcript expression in human thyroid and mucous membrane of small intestine. Real-time PCR and immunohistochemical techniques were used to quantify motilin precursor mRNA and motilin peptide in thyroid tissue samples including adenoma, medullary carcinoma, follicular carcinoma, papillary carcinoma and nodular goiter.

RESULTS(1) The expression of motilin and its precursor mRNA in normal human thyroid was primarily in the thyroid C cells. (2) RT-PCR and Southern blot showed that motilin mRNA expressed in human thyroid was identical to that expressed in duodenum with identical sequence deposited in NCBI Genbank of America. (3) Immunohistochemistry, Western blot research and real-time PCR studies showed that motilin and its precursor mRNA were expressed in normal and tumor tissues of human thyroid. Thyroid tumors (acidophilic adenoma, medullary carcinoma, follicular carcinoma, papillary carcinoma and nodular goiter) showed intense and diffuse immunostaining for motilin peptide. Moreover, the expression of motilin and its precursor mRNA in thyroid medullar carcinoma and acidophilic adenoma were significantly higher than those of normal thyroid tissue (P < 0.05). The expression in thyroid follicular and papillary carcinomas were significantly lower than those of normal thyroid tissue (P < 0.05). There was no difference of the expression between nodular goiter and normal thyroid tissue (P > 0.05).

CONCLUSIONSMotilin peptide and its precursor mRNA are expressed in C cells of human thyroid. The sequence of motilin is identical to that expressed in duodenum from NCBI Genbank of America. The expressions of both motilin and its precursor mRNA in thyroid medullary carcinoma and acidophilic adenoma are significantly increased. In contrast, their expressions in thyroid follicular and papillary carcinomas are significantly decreased. Motilin may regulate physiological functions of the thyroid through parafollicular cells. Motilin may be involved in the pathogenesis of medullary carcinoma and acidophilic adenoma of the thyroid.

Adenocarcinoma, Follicular ; genetics ; Adult ; Aged ; Biomarkers, Tumor ; metabolism ; Carcinoma, Medullary ; genetics ; Carcinoma, Papillary ; genetics ; metabolism ; Female ; Humans ; Intestines ; metabolism ; Male ; Middle Aged ; Motilin ; genetics ; metabolism ; Nervous System Neoplasms ; metabolism ; RNA Precursors ; metabolism ; RNA, Messenger ; metabolism ; Thyroid Gland ; metabolism ; Thyroid Neoplasms ; genetics ; metabolism

PURPOSE: Although follicular thyroid cancer (FTC) has a relatively fair prognosis, distant metastasis sometimes results in poor prognosis and survival. There is little understanding of the mechanisms contributing to the aggressiveness potential of thyroid cancer. We showed that hypoxia inducible factor-1alpha (HIF-1alpha) induced aggressiveness in FTC cells and identified the underlying mechanism of the HIF-1alpha-induced invasive characteristics. MATERIALS AND METHODS: Cells were cultured under controlled hypoxic environments (1% O2) or normoxic conditions. The effect of hypoxia on HIF-1alpha, and epithelial-to-mesenchymal transition (EMT) related markers were evaluated by quantitative real-time PCR, Western blot analysis and immunocytochemistry. Invasion and wound healing assay were conducted to identify functional character of EMT. The involvement of HIF-1alpha and Twist in EMT were studied using gene overexpression or silencing. After orthotopic nude mouse model was established using the cells transfected with lentiviral shHIF-1alpha, tissue analysis was done. RESULTS: Hypoxia induces HIF-1alpha expression and EMT, including typical morphologic changes, cadherin shift, and increased vimentin expression. We showed that overexpression of HIF-1alpha via transfection resulted in the aforementioned changes without hypoxia, and repression of HIF-1alpha with RNA interference suppressed hypoxia-induced HIF-1alpha and EMT. Furthermore, we also observed that Twist expression was regulated by HIF-1alpha. These were confirmed in the orthotopic FTC model. CONCLUSION: Hypoxia induced HIF-1alpha, which in turn induced EMT, resulting in the increased capacity for invasion and migration of cells via regulation of the Twist signal pathway in FTC cells. These findings provide insight into a possible therapeutic strategy to prevent invasive and metastatic FTC.
Adenocarcinoma, Follicular/*genetics/metabolism ; Animals ; Anoxia/*genetics ; Cadherins/genetics ; Epithelial-Mesenchymal Transition/*genetics ; Gene Expression Regulation, Neoplastic ; Hypoxia-Inducible Factor 1, alpha Subunit/*genetics/metabolism ; Lymphokines ; Mice ; Neoplasm Invasiveness ; Phenotype ; Real-Time Polymerase Chain Reaction ; Signal Transduction/drug effects ; Thyroid Neoplasms/*genetics/metabolism ; Transcriptional Activation ; Twist Transcription Factor/*genetics/metabolism ; Vimentin/metabolism

OBJECTIVETo detect the BRAF(V599E) mutation and the RET/PTC chimeric gene in benign and malignant thyroid diseases and to explore their correlation with the clinicopathologic features of papillary thyroid carcinoma (PTC).

METHODSPCR and RT-PCR were employed to detect BRAF(V599E) mutation and RET/PTC chimeric genes in 95 frozen and parraffine-embeded thyroid tissue.

RESULTS(1) BRAF(V599E) mutation was detected only in PTC (56%, 37/66) and had a high prevalence in both classic and tall cell types (70%, 29/41 and 2/3). However, follicular types of PTC and other benign and malignant thyroid diseases were negative for BRAF(V599E) mutation. (2) Fourteen (21.2%) PTC cases expressed RET/PTC chimeric gene. Among them, 5 cases (7.6%) were RET/PTC1 and 9 cases (13.6%) were RET/PTC3 respectively. (3) Statistic data did not show any correlation between the BRAF(V599E) mutation, RET/PTC and clinicopathologic features of PTC (P > 0.05). There was no overlap between BRAF(V599E) mutation and RET/PTC rearrangements.

CONCLUSIONS(1) BRAF(V599E) mutation and RET/PTC rearrangements were unique to PTC. The high prevalence of BRAF(V599E) mutation indicates that it is an important molecular hallmark of PTC. (2) BRAF(V599E) mutation rate was high in classic type PTC and tall cell type inferred that BRAF(V599E) mutation played an important role in their etiopathogenesis. (3) There was no overlap between BRAF(V599E) mutation and RET/PTC rearrangements suggest that they are alternative events in PTC.

Adenocarcinoma, Follicular ; genetics ; pathology ; Adenoma ; genetics ; pathology ; Adolescent ; Adult ; Aged ; Carcinoma, Papillary ; genetics ; pathology ; Diagnosis, Differential ; Female ; Gene Rearrangement ; Hashimoto Disease ; genetics ; pathology ; Humans ; Lymphatic Metastasis ; Male ; Middle Aged ; Oncogene Proteins, Fusion ; genetics ; metabolism ; Point Mutation ; Protein-Tyrosine Kinases ; genetics ; metabolism ; Proto-Oncogene Proteins B-raf ; genetics ; metabolism ; Thyroid Neoplasms ; genetics ; pathology

OBJECTIVETo study immunohistochemical expression of cytokeratin19 (CK19), galectin-3 (Gal-3) and HBME-1 in thyroid lesions and to assess their usefulness as markers in the differential diagnoses of thyroid nodular lesions.

METHODSImmunohistochemical staining was performed on formalin-fixed paraffin-embedded tissue of 21 cases of nodular goiters, 14 cases of toxic goiters, 15 cases of follicular adenomas (FA), 13 cases of follicular carcinomas (FC), 13 cases of follicular variant papillary carcinomas (FVPC) and 48 cases of classic papillary carcinomas (CPC).

RESULTSAll three markers were expressed in the cytoplasm with no or weak expression in benign lesions and diffuse and strong in malignant cases. Positive expressions of CK19, Gal-3 and HBME-1 were present in 11of 21, two of 21, four of 21 in nodular goiters, seven of 14, one of 14, one of 14 in toxic goiters, nine of 15, two of 15, two of 15 in FA, 10 of 13, eight of 13, seven of 13 in FC, 13 of 13, 11 of 13, 12 of 13 in FVPC, and 48 of 48, 45 of 48, 46 of 48 in CPC. The expression rates of the three markers between benign lesions (nodular goiters, toxic goiters and FA) and malignant lesions (FA, FVPC and CPC) were statistically significant. Among the three follicular lesions (FA, FC and FVPC), the differences were statistically significant as well. Nine, seven and six cases were negative for all three markers in nodular goiters, toxic goiters and FA, respectively. Only one case in FC was negative for all three markers, no case was all negative in FVPC and CPC; the rate of one case with two or more positive marker expression in nodular goiters, toxic goiters, FA, FC, FVPC and PC was 14.2% (3/21), 21.43% (3/14), 20.0% (3/15), 69.2% (9/13), 92.3% (12/13), 100.0% (48/48), the differences between benign lesions and malignant lesions and between FA, FC and FVPC were also statistically significant.

CONCLUSIONSImmunohistochemical stains of CK19, Gal-3 and HBME-1, especially when used in combination, can be an important adjunct to the histopathological diagnoses of thyroid lesions.

Adenocarcinoma, Follicular ; chemistry ; diagnosis ; pathology ; Adenoma ; chemistry ; diagnosis ; pathology ; Biomarkers, Tumor ; biosynthesis ; genetics ; Carcinoma, Papillary, Follicular ; pathology ; Diagnosis, Differential ; Galectin 3 ; biosynthesis ; genetics ; Goiter, Nodular ; metabolism ; pathology ; Humans ; Immunohistochemistry ; Keratins ; biosynthesis ; genetics ; Thyroid Neoplasms ; chemistry ; diagnosis ; pathology ; Thyroid Nodule ; chemistry ; diagnosis ; pathology