OBJECTIVE: Histone deacetylase 11 (HDAC11) is a class Ⅳ member of histone deacetylase family, and its role in regulating cancer cell invasion and metastasis remains unclear. We aimed to investigate the role of HDAC11 in regulating the biological behaviors of basal-like breast cancer (BLBC) cells. METHODS: We analyzed the expression of HDAC11 based on Gene Expression Omnibus (GEO) and the Cancer Genome Atlas (TCGA). The effects of HDAC11 on the cell invasion and metastasis were examined using Transwell assay and in a mouse model. The interaction between HDAC11 and Twist was detected with immunoprecipitation. We identified 2 as a target gene of Twist using promoter luciferase assay and chromatin immunoprecipitation assay. RESULTS: HDAC11 was lowly expressed in BLBC cells. HDAC11 overexpression suppressed BLBC cell invasion and their metastasis in nude mice. Mechanistically, HDAC11 directly interacted with Twist protein, antagonized its pro-invasive function and repressed Twist-induced 2 gene transcription. CONCLUSIONS Our data suggest that HDAC11 acts as a negative modulator of invasion and metastasis of BLBC cells.
Animals ; Breast Neoplasms ; Histone Deacetylases ; Mice ; Mice, Nude ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Promoter Regions, Genetic ; Twist-Related Protein 1

OBJECTIVEThe objective of this paper was to study the expression of related protein and Twist transcription factor of epithelial-mesenchymal transition in oral squamous cell carcinoma (OSCC) tissue and the correlations of OSCC and oral squamous cell carcino-metastasis. The paper also investigated the clinical significance of expression on OSCC.

METHODSThe labels of epithelium materialization (E-cadherin and cytokeratin), stromal labels (N-cadherin), transcription factor Twist protein, and mRNA expression in 30 OSCC tissues were investigated via immunohistochemistry and in situ hybridization. The paper also conducted contrast analysis with clinicopathology.

RESULTSImmunization result showed that the expressions of Twist and N-cadherin in the OSCC group were more significant than those of the normal group (P<0.05). The expressions of E-cadherin and keratin in OSCC were significantly lower than those of the normal group (P<0.05). In the moderate- and low-differentiated group of OSCC, the expressions of Twist and N-cadherin were higher than those of the high-differentiated group (P<0.05). The expressions of E-cadherin and keratin were lower than those in the high-differentiated group (P<0.05). In the lymphatic metastasis group, the expressions of Twist and N-cadherin were higher than those of no-lymphatic metastasis group (P<0.05). The expressions of E-cadherin and keratin were lower than those of the no-lymphatic metastasis group (P< 0.05). Results of in situ hybridization showed that the expression of Twist mRNA in the moderate- and low-differentiated groups of OSCC, T3, and T4 groups as well as that of the lymphatic metastasis group were higher than those of the high-differentiated, T1 and T2 groups, and no-separate lymphatic metastasis group, and the differences were statistically significant (P<0.05).

CONCLUSIONEpithelium materialization exists in OSCC tissue. Twist can enhance the invasiveness of tumor cell and promote the infiltration and metastasis of OSCC. The combined detection of Twist, E-cadherin, and N-cadherin expressions can effectively predict and estimate OSCC metastasis.

Cadherins ; Carcinoma, Squamous Cell ; metabolism ; Epithelial Cells ; Epithelial-Mesenchymal Transition ; physiology ; Epithelium ; Humans ; Immunohistochemistry ; Lymphatic Metastasis ; Mouth Neoplasms ; metabolism ; RNA, Messenger ; Twist-Related Protein 1 ; metabolism

Breast Neoplasms ; classification ; genetics ; metabolism ; pathology ; Chromosome Aberrations ; Female ; Gene Deletion ; Humans ; Phyllodes Tumor ; classification ; genetics ; metabolism ; pathology ; Twist-Related Protein 1 ; metabolism ; Wnt Signaling Pathway

OBJECTIVE: To explore the effect of Twist gene on the migration and invasion of human gastric carcinoma cells. METHODS: MKN28 cells, a human gastric carcinoma cell line, were transfected with PcDNA3.1-Twist plasmid by lipofectamine transfecting technique. The transfected cells were selected with geneticin. Expressions of Twist,ecadherin and vimentin protein were detected by Western blot in cells transfected Twist gene. Matrigel invision chambers were performed to analyse the cell migration and invasion. RESULTS: MKN28 cells transfected with PcDNA3.1-Twist plasmid showed stronger intracellular expression of Twist protein than MKN28 cells transfected with PcDNA3.1 and MKN28 cells without transfection. The expression of ecadherin protein in MKN28 cells transfected with PcDNA3.1-Twist plasmid was significantly decreased compared with that in MKN28 cells transfected with PcDNA3.1 and MKN28 cells without the transfection. However, The expression of vimentin protein in MKN28 cells transfected with PcDNA3.1-Twist plasmid was significantly increased compared with that in MKN28 cells transfected with PcDNA3.1 and MKN28 cells without transfection. The migration and invasion ability of Twist+ - MKN28 cells were stronger than that of MKN28 cells transfected with PcDNA3.1 and MKN28 cells without transfection. CONCLUSION Twist gene may promote the migration and invasion ability of gastric carcinoma cells through epithelial mesenchymal transition.
Cadherins ; biosynthesis ; Cell Movement ; genetics ; Humans ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Stomach Neoplasms ; genetics ; metabolism ; pathology ; Transfection ; Tumor Cells, Cultured ; Twist-Related Protein 1 ; biosynthesis ; genetics ; Vimentin ; biosynthesis

Apoptosis ; Breast Neoplasms ; metabolism ; pathology ; Cell Proliferation ; Drug Resistance, Neoplasm ; Epithelial-Mesenchymal Transition ; Female ; Humans ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Neoplastic Stem Cells ; metabolism ; pathology ; Prognosis ; Signal Transduction ; Twist-Related Protein 1 ; metabolism

BACKGROUNDTwist is a highly conserved epithelial-mesenchymal transcription factor that has been reported to be a key factor in tumor malignancy, including lymph node metastasis. It represents the major step of dissemination and serves as a chief prognostic indicator of disease progression. However, the mechanism by which Twist regulates lymph node metastasis remains incompletely understood. Studies on the mechanism of metastasis are thus required for determining appropriate therapeutic strategies.

METHODSImmunohistochemistry for lymphatic vessel endothelial receptor 1 (LYVE-1), Ki-67, Twist, vascular endothelial growth factor C (VEGF-C), and vascular endothelial growth factor receptor 3 (VEGFR-3) was performed to detect lymphatic vessel density (LVD), cell proliferation levels and the expressions of Twist, VEGF-C, and VEGFR-3 were determined from 66 primary supraglottic carcinoma tissue samples from 36 patients with lymph node metastasis (pathological N+, pN+) and 30 patients without metastasis (pathological N0, pN0). Western blotting analysis of the proteins in pN+ and pN0 primary tumors was used to characterize the expressions of Twist, VEGF-C, and VEGFR-3 further.

RESULTSThe LVD was 22.4 ± 10.3 in pN+ patients and 6.8 ± 4.1 in pN0 ones. For Ki-67, the number of proliferous cells in pN+ patients was greater than that in pN0 ones. Both, however, were associated with their clinical nodal stages. In pN+ patients, Twist, VEGF-C, and VEGFR-3 expressions were 86.11% (31/36), 80.56% (29/36), and 58.33% (21/36), respectively. These values were higher than those found for pN0 patients (i.e., 13/30, 11/30, and 7/30, respectively) (P < 0.05). Among the samples with Twist expression, 88.64% were VEGF-C-positive and 59.09% were VEGFR-3-positive. The pN0 counterparts were 4.55% and 9.09%, respectively (P < 0.05). The expressions of Twist, VEGF-C, and VEGFR-3 in pN+ patients obtained through Western blotting analysis were significantly higher than those in pN0 patients, and the levels of VEGF-C and VEGFR-3 were positively correlated with that of Twist.

CONCLUSIONSTwist expression correlates with lymph node metastasis. The mechanism involved in such a correlation may be related to lymphangiogenesis.

Adult ; Aged ; Blotting, Western ; Female ; Humans ; Immunohistochemistry ; Laryngeal Neoplasms ; complications ; metabolism ; Lymphangiogenesis ; genetics ; physiology ; Lymphatic Metastasis ; genetics ; pathology ; Male ; Middle Aged ; Twist-Related Protein 1 ; genetics ; metabolism ; Vascular Endothelial Growth Factor C ; metabolism ; Vascular Endothelial Growth Factor Receptor-3 ; metabolism

OBJECTIVETo investigate whether down-regulation of Twist1 could change sensitivity of nasopharyngeal carcinoma cell line HNE1 to taxol.

METHODSHNE1 cells were transfected with the small interfering RNA (siRNA) expression vector pSuppressor-Retro-Si-Twist, containing the short hairpin RNA (shRNA) sequence targeting the Twist gene-coding region by Fugene 6. Positive clones were then selected in Neomycin (400 microg/ml) for 21 days. The low expressions of Twist1 were examined by real-time reverse-transcription polymerase chain reaction (RT-PCR) and Western blot. Drug sensitivity of si-Twist1 HNE1 to taxol was determined by Annexin V-fluorescein isothiocyanate( FITC)/propidium lodide (PI) double-labeled flow cytometry and detection of DNA ladder. The Effect of Twist1 inactivation on HNE1 cell proliferation was observed by MTT assay and flow cytometry.

RESULTSAnnexin V- FITC-PI assay showed that apoptosis ratio was 40.2% in si-Twist HNE1 after treated with 10 ng/ml taxol, significantly higher than that in the control siRNA group 24.3%. The deference had statistic meaning. After the re-expression of HNE1, apoptosis ratio was 44.80% +/- 4.80% (x +/- s) in low Twist1 protein expression group and that was 27.00% +/- 2.91% in high expression group. The deference had statistic meaning (t = 4.374, P = 0.049). Real time PCR test revealed apoptosis protein bcl-2 expression in si-Twist HNE1 was 0.28 +/- 0.05, significantly lower than that in the control siRNA HNE1 (0.57 +/- 0.08, t = 6.710, P = 0.021), nevertheless, significant bax and bcl-XL changes were not observed (t = 2.000, P = 0.184 and t = 1.502, P = 0.272). MTT and FCM showed that down-regulation of Twist1 did not alter cell proliferation rate (P>0.05).

CONCLUSIONSDown-regulation of Twist1 could increase drug sensitivity of nasopharyngeal carcinoma cell line HNE1 to taxol by inducing apoptosis. These results suggested that Twist1 may be a promising treatment target for nasopharyngeal carcinoma therapy.

Apoptosis ; drug effects ; Cell Line, Tumor ; Down-Regulation ; Gene Expression Regulation, Neoplastic ; Humans ; Nasopharyngeal Neoplasms ; drug therapy ; Nuclear Proteins ; genetics ; Paclitaxel ; pharmacology ; RNA Interference ; RNA, Small Interfering ; Twist-Related Protein 1 ; genetics

OBJECTIVETo investigate the effect of MSX2 interference on epithelial-mesenchymal transitions (EMT) of pancreatic cancer cell line PANC-1.

METHODSThree vectors containing short hairpin RNAs (shRNAs) of MSX2 (shMSX2-1, shMSX2-2, and shMSX2-3) and the empty vector (negative control) were transfected separately into PANC-1 cell line with Lipofectamine2000. Real-time RT-PCR and Western blotting were used to observe changes in the expressions of MSX2, E-cadherin, and vimentin in the cells. CCK-8 assay was used to assess the changes in the cell growth, and wound scratch assay and Transwell assay were employed to evaluate the cell invasion and metastasis after the transfection.

RESULTSAmong the 3 shRNA, shMSX2-1 showed the highest interference efficiency. MSX2 knockdown by the specific shRNA of MSX2 significantly increased E-cadherin expressions, lowered vimentin expressions, and suppressed the invasion, metastasis and proliferation of the cells (P<0.05). MSX2 knockdown also resulted in morphological changes of the cells into cobblestone-like cells in close contact. RT-PCR results revealed significantly reduced mRNA expressions of the transcription factors snail and twist (P<0.05) without affecting slug and zeb1 expressions in the cells with MSX2 knockdown. Conclusion MSX2 knockdown can reverse EMT and induce MET in PANC1 cells, in which process the transcription factors snail and twist may play a role.

Cadherins ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; Epithelial-Mesenchymal Transition ; Homeodomain Proteins ; metabolism ; Humans ; Nuclear Proteins ; metabolism ; Pancreas ; Pancreatic Neoplasms ; pathology ; RNA, Small Interfering ; Snail Family Transcription Factors ; Transcription Factors ; metabolism ; Transfection ; Twist-Related Protein 1 ; metabolism ; Vimentin ; metabolism

OBJECTIVETo investigate the expression of Twist, E-cadherin and N-cadherin in breast carcinoma tissue and to analyse their effects on the breast carcinoma differentiation, size, infiltration and metastasis of the breast carcinoma.

METHODSThe expression of Twist, E-cadherin and N-cadherin in 56 cases of breast invasive ductal carcinoma, 38 cases of invasive lobular carcinoma, 41 cases of carcinoma in situ and 10 cases of normal breast tissue was detected using immunohistochemistry.

RESULTS(1) The expression rate of Twist in three types of breast carcinoma was 46.4% (26/56), 79.0% (30/38) and 26.8% (11/41) respectively, and the expression of Twist in invasive lobular carcinoma was significantly higher than that in invasive ductal carcinoma and carcinoma in situ (P = 0.002, P = 0.000). The expression rate of E-cadherin in three types of breast carcinoma was 78.6% (44/56), 29.0% (11/38) and 80.5% (33/41) respectively, and the expression of E-cadherin in invasive ductal carcinoma and carcinoma in situ was significantly higher than that in invasive lobular carcinoma (P = 0.000, P = 0.000). The expression rate of N-cadherin in three types of breast carcinoma was 53.6% (30/56), 68.4% (26/38) and 31.7% (13/41) respectively, and the expression of N-cadherin in invasive ductal carcinoma and invasive lobular carcinoma was significantly higher than that in carcinoma in situ (P = 0.033, P = 0.001). (2) In all the 135 cases, the expression of Twist was not correlated with that of E-cadherin (P = 0.005, Spearman correlation coefficient = -0.239), however, there was a positive correlation between the expression of Twist and N-cadherin and statistically significant(P = 0.000, Spearman correlation coefficient = 0.319). (3) In the invasive ductal carcinoma, the expression of N-cadherin in poorly-differentiated carcinoma was significantly higher than that of the moderately-or well-differentiated ones (P = 0.004). (4) In the invasive lobular carcinoma, the expression of Twist in cases with lymph node metastasis was significantly higher than that of cases without metastasis (P = 0.037).

CONCLUSIONSTwist, E-cadherin and N-cadherin have different expression patterns in the three kinds of breast carcinoma. The positive expression of Twist was correlated to lymph node metastasis in invasive lobular carcinoma and the positive expression of N-cadherin was correlated to cell the tissue differentiation in invasive ductal carcinoma. Detection of the expression of these biomarkers may provide a valuable reference for the study of breast carcinoma progression, metastasis and for the judgment of the biological behavior of the carcinoma.

Adult ; Aged ; Aged, 80 and over ; Breast Neoplasms ; metabolism ; pathology ; Cadherins ; metabolism ; Carcinoma in Situ ; metabolism ; pathology ; Carcinoma, Ductal, Breast ; metabolism ; pathology ; Carcinoma, Lobular ; metabolism ; pathology ; Female ; Humans ; Immunohistochemistry ; Lymphatic Metastasis ; Middle Aged ; Twist-Related Protein 1 ; metabolism

OBJECTIVETo investigate the role and significance of epithelial-mesenchymal transition (EMT) and its transcriptional regulator Twist1 in the development of the human fetal prostate.

METHODSTwenty-five human fetal prostate specimens at various developmental stages (16-39 weeks) were included in this study. EMT markers, such as E-Cadherin, N-Cadherin and Vimentin, and EMT transcriptional regulator Twist1 were determined by immunohistochemistry, and their relationship with the development of the human fetal prostate was analyzed.

RESULTSE-Cadherin was expressed in the fetal prostate epithelium only, while Vimentin, N-Cadherin and Twist1 in both the epithelium and the stroma. The expression of E-Cadherin gradually increased, but those of Vimentin, N-Cadherin and Twist1 gradually decreased with the gestation stages. No significant changes were observed in the staining patterns of Vimentin, N-Cadherin and Twist1 in the stroma during the whole developmental process.

CONCLUSIONEMT is involved in the development of the human fetal prostate, which may promote epithelial cell motility to form prostatic bud tubules in early gestation stages and boost the differentiation of prostate epithelia in later stages.

Cadherins ; metabolism ; Cell Dedifferentiation ; Epithelial Cells ; metabolism ; Epithelial-Mesenchymal Transition ; Fetal Development ; Humans ; Male ; Mesoderm ; metabolism ; Nuclear Proteins ; metabolism ; Prostate ; embryology ; growth & development ; metabolism ; Twist-Related Protein 1 ; metabolism ; Vimentin ; metabolism