OBJECTIVETo study expression of the zinc finger transcriptional factor Snail in colorectal carcinoma and its significance.

METHODSExpressions of Snail in colorectal carcinoma SW480 and SW620 cells were assayed by immunocytochemistry and immunofluorescent cytochemistry. The paraffin-embedded specimens from 68 cases of colorectal carcinoma and its corresponding adjacent tissues, 33 cases of adenoma and 35 cases of metastatic lymph nodes were also examined for Snail expressions using immunohistochemistry.

RESULTSSnail protein was located mainly in the cell nucleus of SW480 and SW620 cells. The expressions of PRL-3 protein in the specimens of colorectal carcinoma, normal mucosa, adenoma and metastatic lymph nodes were significantly different (Chi(2)=92.852, P=0.000). In the adenoma tissues, the expression was significantly higher than that in normal mucosa (Z=-2.902, P=0.004), the metastatic lymphnodes had significantly higher expressions than the primary colorectal carcinomas (Z=-4.951, P=0.000), which, in turn, showed significantly higher expression than the adenoma tissues (Z=-3.572, P=0.000). Significant correlation of Snail expression was found to the progression and metastasis of colorectal carcinoma (Z=-2.043, P=0.041).

CONCLUSIONThe expression of Snail is significantly correlated to genesis, progression and metastasis of colorectal carcinoma.

Cell Line, Tumor ; Colorectal Neoplasms ; metabolism ; pathology ; Humans ; Immunohistochemistry ; Lymphatic Metastasis ; Snail Family Transcription Factors ; Transcription Factors ; metabolism ; Zinc Fingers

OBJECTIVETo explore the association of Snail expression and Lauren classification of gastric cancer.

METHODSThe protein levels of Snail and E-cadherin were detected by Western blot in N87 (intestinal-type gastric cancer cell line) and AGS(diffuse-type gastric cancer cell line) cell lines and those after transfection of GSK-3β plasmid. The study included a total of 77 patients with primary gastric cancer who underwent curative gastrectomy in the Zhongshan Hospital from February 2000 to December 2005 without any chemotherapy or radiation therapy before surgery. Tissues of gastric cancer specimens were stained using immunohistochemistry to determine Snail expression.

RESULTSSnail expression was low in N78 and high in AGS. E-cadherin expression showed reverse expression pattern. After transfection with GSK-3β, the expression of Snail was significantly suppressed and that of E-cadherin elevated (P<0.01). Different concentrations of GSK-3β inhibitor lithion chloride were used to treat the cell lines and Snail expression was significantly up-regulated in a dose-dependent manner (P<0.01). Snail expression was elevated in 16 out of 21 N78 cell lines, and in 21 out of 56 AGS cell lines, and the difference was statistically significant (P<0.01).

CONCLUSIONThe expression of Snail is closely associated with the Lauren classification of gastric cancer, and it may be a potential marker of the gastric cancer classification.

Cadherins ; metabolism ; Cell Line, Tumor ; Glycogen Synthase Kinase 3 ; genetics ; Humans ; Plasmids ; genetics ; Snail Family Transcription Factors ; Stomach Neoplasms ; classification ; metabolism ; pathology ; Transcription Factors ; metabolism ; Transfection

OBJECTIVE: To explore whether acupuncture combined with moxibustion could inhibit epithelialmesenchymal transition in Crohn's disease by affecting the transforming growth factor β 1 (TGF- β 1)/Smad3/Snail pathway. METHODS: Sixty-three patients with Crohn's disease were randomly divided into an observation group (31 cases) receiving moxibustion at 43 °C combined with acupuncture, and a control group (32 cases) receiving moxibustion at 37 °C combined with sham acupuncture using a random number table. Patients were treated for 12 weeks. Crohn's Disease Activity Index (CDAI) was used to evaluate disease activity. Hematoxylin-eosin staining and transmission electron microscopy were utilized to observe the morphological and ultrastructural changes. Immunohistochemistry was used to detect the expression of transforming growth factor β 1 (TGF-β 1), T β R1, T β R2, Smad3, Snail, E-cadherin and fibronectin in intestinal mucosal tissues. RESULTS: The decrease of the CDAI score, morphological and ultrastructural changes were more significant in observation group. The expression levels of TGF- β 1, Tβ R2, Smad3, and Snail in the observation group were significantly lower than those before the treatment (P<0.05 or P<0.01). After treatment, the expression levels of TGF-β 1, TβR2, and Snail in the observation group were significantly lower than those in the control group (all P<0.05); compared with the control group, the expression of fibronectin in the observation group was significantly decreased, and the expression of E-cadherin was significantly increased (all P<0.05). CONCLUSIONS Moxibustion at 43 °C combined with acupuncture may suppress TGF-β 1/Smad3/Snail pathway-mediated epithelial-mesenchymal transition of intestinal epithelial cells in Crohn's disease patients by inhibiting the expression levels of TGF-β 1, Tβ R2, Smad3, and Snail. (Registration No. ChiCTR-IIR-16007751).
Acupuncture Therapy ; Cadherins/metabolism* ; Crohn Disease/therapy* ; Epithelial-Mesenchymal Transition ; Fibronectins/metabolism* ; Humans ; Moxibustion ; Smad3 Protein/metabolism* ; Snail Family Transcription Factors/metabolism* ; Transforming Growth Factor beta1/metabolism*

OBJECTIVETo explore expressions of CD133, E-cadherin and Snail in human epithelial ovarian cancer (EOC) and elucidate their relationship with the clinicopathologic features and prognosis of the patients.

METHODSThe expression of CD133, E-cadherin and Snail were detected by immunohistochemical staining in 150 specimens of EOC and 50 specimens of benign ovarian epithelial tumor tissues.

RESULTSThe positivity rates of CD133, E-cadherin and Snail protein in EOC were 58.7%, 60.7% and 32.7%, respectively, significantly different from the rates in benign epithelial tumor tissues (10%, 8.0%, and 70%, respectively; P<0.05). The expressions of CD133, E-cadherin and Snail in EOC were significantly correlated with abdominal organ and lymphnode metastases and FIGO stage (P<0.01). E-cadherin expression was inversely correlated with Snail and CD133 expression (r=-0.545 and -0.570, P<0.01), and the latter two were positively correlated (r=0.599, P<0.01). Overexpressions of CD133 and Snail and a decreased expression of E-cadherin were all related to a poor prognosis of the patients (P<0.05). FIGO stage and expressions of CD133, E-cadherin and Snail were all independent prognostic factors of EOC (P<0.05).

CONCLUSIONThe expressions of CD133, E-cadherin and Snail are related to lymph node metastasis, clinical stage, and prognosis of EOC. Combined detection of these indexes provides important evidence for predicting the progression and prognosis of EOC.

AC133 Antigen ; Antigens, CD ; metabolism ; Cadherins ; metabolism ; Disease Progression ; Female ; Glycoproteins ; metabolism ; Humans ; Lymphatic Metastasis ; Neoplasms, Glandular and Epithelial ; metabolism ; pathology ; Ovarian Neoplasms ; metabolism ; pathology ; Peptides ; metabolism ; Prognosis ; Snail Family Transcription Factors ; Transcription Factors ; metabolism

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 explore the expression of Snail and Slug in primary cervical squamous cell carcinoma (CSCC) and their relationship with KAI1 expression.

METHODSThe expressions of Snail, Slug, and KAI1 proteins were examined by immunohistochemistry in 154 specimens of CSCC tissues, 50 specimens of cervical intraepithelial neoplasm (CIN), and 40 specimens of normal cervical tissues.

RESULTSThe positivity rates of Snail, Slug, and KAI1 expression were 0%, 2.5%, and 95.0% in normal cervical tissues, 32.0%, 34.0% and 64.0% in CIN tissues, and 66.2%, 66.9%, and 43.5% in CSCC tissues, respectively, showing significant differences in the rates among the 3 groups (P<0.05). The expressions of Snail, Slug, and KAI1 were significantly correlated with the histological grades of the tumor, depth of invasion, lymph node metastasis, International Federation of Gynecology and Obstetrics (FIGO) stages, and postoperative survival time (P<0.05). The expressions of Snail and Slug were positively correlated (r=0.752, P<0.001), and both of them were negatively correlated with the expression of KAI1 (P<0.001). Kaplan-Meier analysis showed that patients positive for Snail and Slug had significantly lower survival rates than the negative patients (P<0.001), while a positive expression of KAI1 was associated with a higher survival rate of the patients. Cox regression analysis identified Snail, KAI1, and FIGO stage as independent factors that affected the outcomes of CSCC (P<0.05).

CONCLUSIONThe expressions of Snail, Slug, and KAI1 are related to the tumor grade, FIGO stage, invasive depth, lymph node metastasis, and prognosis of CSCC, and their combined detection can help estimate the outcomes of the patients.

Carcinoma, Squamous Cell ; metabolism ; pathology ; Cervical Intraepithelial Neoplasia ; metabolism ; pathology ; Female ; Humans ; Immunohistochemistry ; Kangai-1 Protein ; metabolism ; Kaplan-Meier Estimate ; Lymphatic Metastasis ; Neoplasm Staging ; Prognosis ; Snail Family Transcription Factors ; Survival Rate ; Transcription Factors ; metabolism ; Uterine Cervical Neoplasms ; metabolism ; pathology

The present study was aimed to explore the expressions of transforming growth factor-β1 (TGF-β1) and Snail1 in renal tissues of diabetic rats, and their role in tubular epithelial-mesenchymal transition (TEMT). Induced diabetic rats were randomly divided into 2-, 4-, 8-, 12-, 16-, 20-, 24-week and 16wA, 20wA, 24wA groups. The rats in 16wA, 20wA and 24wA groups were treated with insulin to control blood glucose to the normal level from the 13th week. The age-matched rats were set as controls. Blood glucose, 24-hour urine protein, serum creatinine (Scr), kidney index of rats were measured. PAS staining was used to observe the renal pathological changes. Immunohistochemical staining and (or) Western blot were employed to determine the expressions of TGF-β1, Snail1, E-cadherin, α-smooth muscle actin (α-SMA) and fibronectin (FN) proteins. The expressions of Snail1 and E-cadherin mRNAs in renal cortex were examined by RT-PCR. Blood glucose, 24-hour urine protein, Scr and kidney index increased remarkably in diabetic rats as compared with those in the control groups (P<0.05, P<0.01) and insulin-treated rats (P<0.01). TGF-β1 and Snail1 protein expressions could not be detected by immunohistochemical staining in the normal renal tissues, however, the strongly positive staining was observed in diabetic rat renal tubules. A time-dependent loss of TGF-β1 and Snail1 expressions was detected in the kidney of insulin-treated rats. In diabetic rats tubular α-SMA positive staining was seen at the 16th week. E-cadherin expression was lost in diabetic rats. The expressions of TGF-β1, Snail1 proteins and Snail1 mRNA were significantly up-regulated in diabetic rats, while down-regulated in insulin-treated rats (P<0.01). The expressions of E-cadherin protein and mRNA in the cortex were contrary to the expressions of TGF-β1 and Snail1. Therefore, TGF-β1 and Snail1 are possibly involved in the pathogenesis of TEMT in diabetic nephropathy rats.
Animals ; Diabetes Mellitus, Experimental ; metabolism ; Diabetic Nephropathies ; metabolism ; Down-Regulation ; Epithelial-Mesenchymal Transition ; Kidney ; pathology ; Kidney Tubules ; metabolism ; Rats ; Snail Family Transcription Factors ; Transcription Factors ; metabolism ; Transforming Growth Factor beta1 ; metabolism

OBJECTIVETo identify the region in PRL-3 gene promoter where the transcriptional factor Snail can bind.

METHODSPRL-3 promoter and the possible binding sites of the transcription factors were analyzed by bioinformatical methods. Chromatin immunoprecipitation and PCR were performed using the antibody specific for Snail to verify the binding of Snail to PRL-3 promoter.

RESULTSAccording to the prediction by TRED, a promoter prediction software, PRL-3 gene promoter was located between -700 bp to 299 bp of PRL-3 gene. Many possible transcription factor binding sites such as for Snail, n-MYC, ARNT, E74A, NF-kappaB, NRF-2 and AML-1 were predicted by Consite, a promoter analysis web system. Interestingly, a 5'-CACCTG-3' core sequence and other related sequences of Snail binding sites were found in the promoter region of PRL-3 genes by Consite software. Two regions in PRL-3 promoter were validated to allow binding of Snail by chromatin immunoprecipitation analysis of SW480 cells.

CONCLUSIONSSnail regulates the activity of PRL-3 gene by binding to the promoter of PRL-3 gene in SW480 cells.

Base Sequence ; Binding Sites ; Cell Line, Tumor ; Computational Biology ; Humans ; Molecular Sequence Data ; Neoplasm Proteins ; metabolism ; Promoter Regions, Genetic ; Protein Tyrosine Phosphatases ; metabolism ; Snail Family Transcription Factors ; Software ; Transcription Factors ; metabolism

OBJECTIVEMalignant transformation of epithelial cell frequently coincides with loss of E-cadherin. Here we study the expression of Snail and E-cadherin and correlate their expression with cell differentiation and in vitro invasion.

METHODSThe expression and localization of Snail and E-cadherin were studied by Northern blot and laser confocal microscopy in two normal cell lines (MDCK, NIH 3T3) and six carcinoma cell lines (A431, MCF-7, MDA-MB-453, HepG2, MDA-MB-435s, MDA-MB-231). Boyden chamber assay was done to detect the invasive ability of cells in vitro.

RESULTSSnail mRNA and protein were detected in fibroblasts NIH 3T3 and poorly differentiated carcinoma cell lines HepG2, MDA-MB-435s and MDA-MB-231. On the contrary, E-cadherin mRNA and protein were detected in normal epithelial cell line MDCK and well differentiated carcinoma cell lines A431 and MDA-MB-453. In MCF-7 cells, Snail and E-cadherin expressions were revealed both at mRNA and protein levels. The cells with higher expression of Snail had stronger ability of invasion than those with lower expression of Snail.

CONCLUSIONThere is an inverse correlation between Snail and E-cadherin expressions and their expressions are correlated with cell differentiation and tumor invasiveness.

3T3 Cells ; metabolism ; Animals ; Cadherins ; biosynthesis ; genetics ; Cell Differentiation ; Cell Line ; Cell Line, Tumor ; Cell Transformation, Neoplastic ; metabolism ; Dogs ; Epithelial Cells ; cytology ; metabolism ; Humans ; Mice ; Neoplasm Invasiveness ; Snail Family Transcription Factors ; Transcription Factors ; biosynthesis ; genetics

In order to investigate the transfer and expression of Snail gene in human bone mesenchymal stem cells (MSCs) and to study effects of Snail gene modification on the CXCR4 expression of human MSCs and their capacity of migration to SDF-1 in vitro, the plasmid PCAGGSneo-Snail-HA or the control vector of PCAGGSneo was transferred into the cells. Fluorescence activated cell sorting analysis, immunofluorescence staining and RT-PCR were used to study the expression of CXCR4 by MSCs. Chemotaxis assays were performed to evaluate the migratory capacity of MSCs-Sna and MSCs-neo to SDF-1 in vitro. For the blocking assay, CXCR4 blocking antibody was added into cell culture. CXCR4 expression was higher in MSCs-Sna than that in MSCs-neo (P < 0.05). Chemotaxis assays showed that SDF-1alpha stimulated migratory activity of MSCs-Sna more than MSCs-neo in vitro (P < 0.05). Moreover, the SDF-1alpha-induced migratory activity of MSCs-Sna was inhibited in a concentration-dependent manner by a CXCR4-blocking antibody. It was concluded that Snail enhanced expression of CXCR4 in MSCs, providing a plausible mechanism for Snail-mediated MSCs transmigration to damaged tissues in vivo where SDF-1 has been shown to be up-regulated as part of injury responses.
Bone Marrow Cells ; cytology ; Cell Movement ; genetics ; Cells, Cultured ; Chemokine CXCL12 ; metabolism ; Humans ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Receptors, CXCR4 ; genetics ; metabolism ; Snail Family Transcription Factors ; Transcription Factors ; genetics ; Transduction, Genetic