BACKGROUND: Epithelial-mesenchymal transition (EMT) is critical for morphogenesis during embryonic development and is also implicated in the conversion of early-stage tumors into invasive malignancies. Recently, Twist has been identified to play an important role in EMTmediated metastatic progression of several types of human cancer. The present study examined the expression of Twist and evaluated its clinicopathologic significance in urothelial carcinoma of upper urinary tract. METHODS: Immunohistochemical staining for Twist expression was performed on 70 upper urinary tract urothelial carcinomas (UUT-UCs) using tissue microarray. RESULTS: Immunohistochemical staining for Twist was positive in 31/70 cases (44.3%) of UUT-UCs. Twist expression was associated with high-grade and advanced-stage (ISUP grade, p<0.01; stage, p=0.045). The patients with Twist positive-tumors revealed lower disease free survival rate than those with Twist negative-tumors (p<0.01). The overall survival for patients with Twist positive-tumors was slightly worse than the patients with Twist negative- tumors, but the difference was not statistically significant (p=0.12). CONCLUSION: Our results suggest that Twist is a novel marker for advanced UUT-UC.
Carcinoma, Transitional Cell ; Disease-Free Survival* ; Embryonic Development ; Epithelial-Mesenchymal Transition ; Female ; Humans ; Morphogenesis ; Pregnancy ; Twist Transcription Factor ; Urinary Tract*

BACKGROUND: Epithelial-mesenchymal transition (EMT) is associated with tumor hypoxia. EMT is regulated, in part, by the action of TWIST, which inhibits of E-cadherin expression and may interfere with the p53 tumor-suppressor pathway. METHODS: We examined the expression of TWIST, E-cadherin, hypoxia-inducible factor 1alpha (HIF1alpha), and p53 by immunohistochemistry in 123 cases of ovarian epithelial cancers (OEC) to evaluate the role of TWIST in OEC. We assessed the association between protein expression and clinicopathologic parameters. RESULTS: The expression of TWIST, E-cadherin, HIF1alpha, and p53 proteins was found in 28.5%, 51.2%, 35.0%, and 29.3% of cases, respectively. TWIST expression was associated with higher histologic grade and unfavorable survival. TWIST expression was correlated with HIF1alpha expression and reduced E-cadherin expression. The altered HIF1alpha/TWIST/E-cadherin pathway was associated with lower overall survival (OS), while the co-expression of TWIST and p53 was correlated with lower progression-free survival. In the multivariate analyses, TWIST expression was an independent prognostic factor for OS. CONCLUSIONS: Our data imply that TWIST expression could be a useful predictor of unfavorable prognosis for OEC. TWIST may affect the p53 tumor-suppressor pathway. Moreover, hypoxia-mediated EMT, which involves the HIF1alpha/TWIST/E-cadherin pathway may play an important role in the progression of OEC.
Anoxia ; Cadherins ; Disease-Free Survival ; Epithelial-Mesenchymal Transition ; Immunohistochemistry ; Multivariate Analysis ; Prognosis ; Tumor Suppressor Protein p53 ; Twist Transcription Factor

In order to investigate the role of Twist gene in the metastasis of hepatocellular carcinoma (HCC), total RNA was respectively extracted from three HCC cell strains with different metastatic potentials, HepG2, MHCC-97L and MHCC-97H. The first strand cDNA was synthesized by reverse transcription, which was then used as template to perform fluorescent quantitative polymerase chain reaction (FQ-PCR). The quantity of Twist gene expression was normalized by that of the housekeeping gene, GAPDH for each sample. ANOVA was used to estimate the relationship between Twist gene and metastasis potential of HCC. The results showed that the normalized initial cDNA concentrations of Twist gene in HepG2, MHCC-97L and MHCC-97H were (9.45+/-0.25)x10(-4), (1.82+/-0.41)x10(-3), (3.06+/-0.62)x10(-3), respectively. FQ-PCR revealed significant differences in the expression level of Twist among HCC cell strains with different metastatic potentials. It was concluded that high expression level of Twist was closely associated with more aggressive behaviors of HCC. Twist provides a novel indicator for HCC metastasis.
Carcinoma, Hepatocellular/*metabolism ; Cell Line, Tumor ; DNA, Complementary/metabolism ; Dose-Response Relationship, Drug ; Gene Expression Regulation, Neoplastic ; Liver Neoplasms/*metabolism ; Neoplasm Metastasis ; Nuclear Proteins/*biosynthesis ; Polymerase Chain Reaction ; RNA/metabolism ; Twist Transcription Factor/*biosynthesis

Speckle tracking imaging (STI) was employed to investigate the effect of right ventricular (RV) volume and pressure overload on left ventricular (LV) rotation and twist in 35 patients with atrial septal defect (ASD), 18 of which with pulmonary hypertension, and 21 healthy subjects serving as controls. The peak rotations of 6 segments at the basal and apical short-axises and the average peak rotation and interval time of the 6 segments in the opposite direction during early systolic phase were measured respectively. LV twist versus time profile was drawn and the peak twist and time to peak twist were calculated. LV ejection fraction (EF) was measured by Biplane Simpson. Compared to ASD patients without pulmonary hypertension and healthy subjects, the peak rotations of posterior, inferior and postsept walls at the basal level were lower (P<0.05), and the average counterclockwise peak rotation of 6 segments at the basal level during early systolic phase was higher (P<0.05), and the average interval time was delayed (P<0.05). LV peak twist was also lower (P<0.05), and had a significant negative correlation with pulmonary arterial systolic pressure (r=-0.57, P=0.001). No significant differences were found in LVEF among the three groups. It was suggested that although RV volume overload due to ASD has no significant effects on LV rotation and twist, LV peak twist is lower in ASD patients with pulmonary hypertension. Thus LV twist may serve as a new indicator of the presence of pulmonary hypertension in ASD patients.
Cardiology/methods ; Echocardiography/methods ; Echocardiography, Doppler/methods ; Heart Septal Defects, Atrial/metabolism ; Heart Septal Defects, Atrial/*pathology ; Heart Ventricles/pathology ; Hypertension ; Nuclear Proteins/*metabolism ; Systole ; Twist Transcription Factor/*metabolism ; Ventricular Function, Left

Cell Hypoxia ; Epithelial-Mesenchymal Transition ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Neoplasms ; metabolism ; pathology ; Receptors, Notch ; metabolism ; Signal Transduction ; Snail Family Transcription Factors ; Transcription Factors ; metabolism ; Transforming Growth Factor beta ; metabolism ; Twist-Related Protein 1 ; metabolism ; Wnt Proteins ; 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

OBJECTIVE: To evaluate the relation of EB virus latent membrane protein-1 (LMP-1) and the phenotype of epithelial-mesenchymal transition (EMT) and cervical lymph node metastasis in nasopharyngeal carcinoma. METHOD: Based on histopathology and MRI imaging, nasopharyngeal biopsy tissues from 88 patients with nasopharyngeal carcinoma were divided into 3 groups: pathologic metastasis (18), MRI metastasis(40) and without metastasis (30). The expressions of LMP-1, STAT3, Twist, E-Cadherin and Vimentin were examined immunohistochemically in biopsy tissues. RESULT: LMP-1 expression was found in 35 of 88 biopsy tissues with a positive rate of 38.7%. The positive rates of LMP-1 in groups of pathologic metastasis, MRI metastasis and without metastasis were 38.9% (7/18), 47.5% (19/40) and 30.0% (9/30), respectively, and significant difference were not found among three groups. The expression of LMP-1 was positively correlated to both expressions of Twist and Vimentin (r = 0.276 and 0.282, are P < 0.01), but not to both expressions of STAT3 and E-Cadherin. The positive expressions or abnormal expression of STAT3, Twist, Vimentin and E-Cadherin were found in 57 of 88 (64.8%), 48 of 88 (54.5%), 22 of 88 (20.0%)and 53 of 88 (60.2%), respectively. Significant differences in the expression of STAT3, Twist, Vimentin and E-Cadherin were all found among groups of pathologic metastasis, MRI metastasis and without metastasis, respectively (are P < 0.05). The expression of STAT3 was positively correlated to both expressions of Twist and Vimentin (r = 0.712 and 0.316, P < 0.01). CONCLUSION EMT plays important role in cervical lymph node metastasis in nasopharyngeal carcinoma. LMP-1 may be only as one of upstream factors associated with the EMT, but not the decisive factor for cervical lymph node metastasis.
Adult ; Aged ; Aged, 80 and over ; Cadherins ; metabolism ; Epithelial-Mesenchymal Transition ; Female ; Herpesvirus 4, Human ; metabolism ; Humans ; Lymphatic Metastasis ; Male ; Middle Aged ; Nasopharyngeal Neoplasms ; pathology ; virology ; Neck ; pathology ; Nuclear Proteins ; metabolism ; STAT3 Transcription Factor ; metabolism ; Twist-Related Protein 1 ; metabolism ; Vimentin ; metabolism ; Viral Matrix Proteins ; metabolism ; Young Adult