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1.
Acta cir. bras ; 36(10): e361007, 2021. tab, graf
Article in English | LILACS, VETINDEX | ID: biblio-1349866

ABSTRACT

ABSTRACT Purpose: To evaluate the effects of sucralfate enemas in tissue contents of E-cadherin and ?-catenin in an experimental diversion colitis. Methods: Thirty-six male Wistar rats were submitted to a proximal colostomy and a distal mucous fistula. They were allocated into three groups: first group received daily saline enemas (2 mL/day) and the two other groups daily enemas with sucralfate at dosage of 1 or 2 g/kg/day, respectively. Six animals of each group were euthanized after two weeks and six animals after four weeks. The inflammation of the excluded mucosa was evaluated by histological analysis. The oxidative damage was quantified by measurement of malondialdehyde tissue levels. The expression of E-cadherin and ?-catenin was identified by immunohistochemistry, and its contents were quantified by computer-assisted image analysis. Results: Sucralfate enemas reduced inflammation in animals subjected to treatment with 2 g/kg/day by four weeks, and the levels of oxidative damage in mucosa without fecal stream irrespective of concentration and time of intervention. E-cadherin and ?-catenin content increased in segments without fecal stream in those animals subjected to treatment with sucralfate. Conclusions: Sucralfate reduces the inflammation and oxidative stress and increases the tissue content of E-cadherin and ?-catenin in colonic mucosa devoid to the fecal stream.


Subject(s)
Humans , Animals , Rats , Sucralfate/metabolism , Catenins/metabolism , Cadherins/metabolism , Rats, Wistar , Oxidative Stress , Enema , Intestinal Mucosa/metabolism
2.
Journal of Experimental Hematology ; (6): 1367-1373, 2019.
Article in Chinese | WPRIM | ID: wpr-775712

ABSTRACT

OBJECTIVE@#To study the effects of miR-221 on the biological activity of childhood acute lymphoblastic leukemia cells and its mechanism.@*METHODS@#Bone marrow mononuclear cells (BMNC) were isolated from bone marrow samples of ALL children diagnosed in our hospital from May 2018 to November 2018. The cells were divided into control group, miR-221-NC group and miR-221 group. After transfection according to the instructions of Lipofectamine 2000 kit, the levels of miR-221 in each group were detected by RT-PCR. Flow cytometry was used to detect the effects of miR-221 on cell cycle and apoptosis. Transwell assay was used to detect cell migration and invasion. Western blot was used to detect the effects of miR-221 on proliferating cell nuclear antigen (PCNA), Caspase 3, Cyclin D1 and MMP-9 proteins in BMNC. Luciferase reporter gene assay was used to detect the targeting relationship between miR-221 and Wnt gene.@*RESULT@#The expression level of miR-221 in the miR-221 group was significantly higher than that in the control group and the miR-221-NC group (P<0.05). MTT assay showed that, after transfection for 2, 3, 4 and 5 days, the cell proliferation level in miR-221 group was significantly lower than that in the control group and the miR-221-NC group (P<0.05). The cell ratio of G/G phase was (73.25±8.1)% in the miR-221 group, which was significantly higher than that in the control group and the miR-221-NC group (P<0.05); moreover, the cell ratio of S phase in the miR-221 group was (12.37±1.6)%,which was significantly lower than that in the control group and the miR-221-NC group (P<0.05). The percentage of apoptotic cells in the miR-221 group was (24.68±3.87)%, which was significantly higher than that in the control group and the miR-221-NC group (P<0.05). Transwell cell invasion experiment showed that the number of invasive cells in the miR-221 group was 23.42±3.62, which was significantly lower than that in the control group and the miR-221-NC group (P<0.05). Transwell cell migration assay showed that the number of migrating cells in the miR-221 group was 34.86±5.32, which was significantly lower than that in the control group and the miR-221-NC group (P<0.05). The relative level of PCNA, Cyclin D1 and MMP-9 in the miR-221 group was 0.26±0.03, 0.17±3.61 and 0.14±0.02, respectively, which was significantly lower than those in the control group and the miR-221-NC group (P<0.05), while the relative level of Caspase-3 in the miR-221 group was 0.37±0.05, which was significantly higher than that in the control group and the miR-221-NC group (P<0.05). Luciferase reporter assay showed that the activity of luciferase in Wnt wild type plasmid was significantly inhibited by miR-221 (P<0.05).@*CONCLUSION@#miR-221 can inhibit the proliferation, migration and invasion of BMNC, moreover can promote cell apoptosis, which may be related with the inhibition of Wnt/β- catenin signaling pathway.


Subject(s)
Catenins , Cell Line, Tumor , Cell Proliferation , Child , Humans , MicroRNAs , Precursor Cell Lymphoblastic Leukemia-Lymphoma , Wnt Signaling Pathway
3.
Biol. Res ; 51: 31, 2018. graf
Article in English | LILACS | ID: biblio-983936

ABSTRACT

BACKGROUND: miR-214 was demonstrated to be upregulated in models of renal disease and promoted fibrosis in renal injury independent of TGF-ß signaling in vivo. However, the detailed role of miR-214 in acute kidney injury (AKI) and its underlying mechanism are still largely unknown. METHODS: In this study, an I/R-induced rat AKI model and a hypoxia-induced NRK-52E cell model were used to study AKI. The concentrations of kidney injury markers serum creatinine, blood urea nitrogen, and kidney injury molecule-1 were measured. The expressions of miR-214, tumor necrosis factor-α, interleukin (IL)-1ß, IL-6, were detected by RT-qPCR. The protein levels of Bcl-2, Bax, Dickkopf-related protein 3, ß-catenin, c-myc, and cyclinD1 were determined by western blot. Cell apoptosis and caspase 3 activity were evaluated by flow cytometry analysis and caspase 3 activity assay, respectively. Luciferase reporter assay was used to confirm the interaction between miR-214 and Dkk3. RESULTS: miR-214 expression was induced in ischemia-reperfusion (I/R)-induced AKI rat and hypoxic incubation of NRK-52E cells. Overexpression of miR-214 alleviated hypoxia-induced NRK-52E cell apoptosis while inhibition of miR-214 expression exerted the opposite effect. Dkk3 was identified as a target of miR-214. Anti-miR-214 abolished the inhibitory effects of DKK3 knockdown on hypoxia-induced NRK-52E cell apoptosis by inactivation of Wnt/ß-catenin signaling. Moreover, miR-214 ameliorated AKI in vivo by inhibiting apoptosis and fibrosis through targeting Dkk3 and activating Wnt/ß -catenin pathway. CONCLUSION: miR-214 ameliorates AKI by inhibiting apoptosis through targeting Dkk3 and activating Wnt/ß -catenin signaling pathway, offering the possibility of miR-214 in the therapy of ischemic AKI.


Subject(s)
Animals , Male , Rats , Intercellular Signaling Peptides and Proteins/metabolism , MicroRNAs/metabolism , Catenins/metabolism , Acute Kidney Injury/metabolism , Wnt Signaling Pathway/genetics , Rats, Sprague-Dawley , Chemokines , Intercellular Signaling Peptides and Proteins/genetics , MicroRNAs/genetics , Adaptor Proteins, Signal Transducing , Cell Proliferation , Disease Models, Animal , Catenins/genetics , Acute Kidney Injury/chemically induced
4.
Chinese Medical Journal ; (24): 804-812, 2018.
Article in English | WPRIM | ID: wpr-687035

ABSTRACT

<p><b>Background</b>Ventilator-induced lung injury (VILI) is commonly associated with barrier dysfunction and inflammation reaction. Glutamine could ameliorate VILI, but its role has not been fully elucidated. This study examined the relationship between inflammatory cytokines (interleukin [IL]-6, tumor necrosis factor [TNF]-α, and IL-10) and adherens junctions (E-cadherin, p120-catenin), which were ameliorated by glutamine in VILI, both in vitro and in vivo.</p><p><b>Methods</b>For the in vivo study, 30 healthy C57BL/6 mice weighing 25-30 g were randomly divided into five groups with random number table (n = 6 in each group): control (Group C); low tidal volume (Group L); low tidal volume + glutamine (Group L + G); high tidal volume (Group H); and high tidal volume + glutamine (Group H + G). Mice in all groups, except Group C, underwent mechanical ventilation for 4 h. For the in vitro study, mouse lung epithelial 12 (MLE-12) cells pretreated with glutamine underwent cyclic stretching at 20% for 4 h. Cell lysate and lung tissue were obtained to detect the junction proteins, inflammatory cytokines, and lung pathological changes by the Western blotting, cytokine assay, hematoxylin and eosin staining, and immunofluorescence.</p><p><b>Results</b>In vivo, compared with Group C, total cell counts (t = -28.182, P < 0.01), the percentage of neutrophils (t = -28.095, P < 0.01), IL-6 (t = -28.296, P < 0.01), and TNF-α (t = -19.812, P < 0.01) in bronchoalveolar lavage (BAL) fluid, lung injury scores (t = -6.708, P < 0.01), and the wet-to-dry ratio (t = -15.595, P < 0.01) were increased in Group H; IL-10 in BAL fluid (t = 9.093, P < 0.01) and the expression of E-cadherin (t = 10.044, P < 0.01) and p120-catenin (t = 13.218, P < 0.01) were decreased in Group H. Compared with Group H, total cell counts (t = 14.844, P < 0.01), the percentage of neutrophils (t = 18.077, P < 0.01), IL-6 (t = 18.007, P < 0.01), and TNF-α (t = 10.171, P < 0.01) in BAL fluid were decreased in Group H + G; IL-10 in BAL fluid (t = -7.531, P < 0.01) and the expression of E-cadherin (t = -14.814, P < 0.01) and p120-catenin (t = -9.114, P < 0.01) were increased in Group H + G. In vitro, compared with the nonstretching group, the levels of IL-6 (t = -21.111, P < 0.01) and TNF-α (t = -15.270, P < 0.01) were increased in the 20% cyclic stretching group; the levels of IL-10 (t = 5.450, P < 0.01) and the expression of E-cadherin (t = 17.736, P < 0.01) and p120-catenin (t = 16.136, P < 0.01) were decreased in the 20% cyclic stretching group. Compared with the stretching group, the levels of IL-6 (t = 11.818, P < 0.01) and TNF-α (t = 8.631, P < 0.01) decreased in the glutamine group; the levels of IL-10 (t = -3.203, P < 0.05) and the expression of E-cadherin (t = -13.567, P < 0.01) and p120-catenin (t = -10.013, P < 0.01) were increased in the glutamine group.</p><p><b>Conclusions</b>High tidal volume mechanical ventilation and 20% cyclic stretching could cause VILI. Glutamine regulates VILI by improving cytokines and increasing the adherens junctions, protein E-cadherin and p120-catenin, to enhance the epithelial barrier function.</p>


Subject(s)
Animals , Cadherins , Metabolism , Catenins , Metabolism , Glutamine , Metabolism , Inflammation , Metabolism , Interleukin-6 , Metabolism , Lung , Metabolism , Pathology , Mice , Mice, Inbred C57BL , Ventilator-Induced Lung Injury , Allergy and Immunology , Metabolism
5.
Article in Chinese | WPRIM | ID: wpr-357498

ABSTRACT

<p><b>OBJECTIVE</b>The main goal is to investigate the role of P120-catenin (P120ctn) in cadherin switching, as well as migration and invasion, of oral squamous cell cancer (OSCC) cells.</p><p><b>METHODS</b>The plasmid pGFP-V-RS-P120ctn shRNA was used to transfect TSCCA cells and significantly reduce the expression of P120ctn in these cells. Real-time fluorescent quantitative polymerase chain reaction and Western blot were conducted to determine the mRNA and protein expression levels of P120ctn, E-cadherin (E-cad), and N-cadherin (N-cad). By contrast, the Transwell cell invasion and cell migration assay was used to determine the invasion and migration capacities before and after the transfection.</p><p><b>RESULTS</b>After the plasmid pGFP-V-RS-P120ctn shRNA was transfected into the TSCCA cells, we found that as the P120ctn expression significantly decreased, E-cad mRNA and protein expression decreased significantly. Moreover, N-cad mRNA and protein expression increased significantly (P<0.05). Lastly, the cell migration and invasion capacities were augmented significantly (P<0.05).</p><p><b>CONCLUSIONS</b>In OSCC cells, P120ctn may be involved in cadherin switching and promote metastasis and invasion.</p>


Subject(s)
Cadherins , Carcinoma, Squamous Cell , Catenins , Cell Line, Tumor , Cell Movement , Humans , Mouth Neoplasms , Neoplasm Invasiveness , Neoplasm Metastasis , Transfection
6.
Article in English | WPRIM | ID: wpr-250409

ABSTRACT

This study was aimed to investigate the expressions of E-cadherin, p120ctn, β-catenin and NF-κB in ulcerative colitis (UC) tissues and the implications of their expressions in the pathogenesis of UC. The expressions of E-cadherin, p120ctn, β-catenin and NF-κB were detected by immunohistochemistry, and those of p120ctn and NF-κB by Western blotting in 23 cases of UC and 17 cases of normal colonic tissues. The relationship between the expression of E-cadherin or NF-κB and that of p120ctn was analyzed by Spearman rank correlation analysis. The results showed that in UC and normal colonic groups, the abnormal expression rate of E-cadherin, p120ctn, β-catenin, and NF-κB was 52.2% vs. 0 (P<0.05), 73.9% vs. 23.5% (P<0.05), 65.2% vs. 17.6% (P<0.05) and 78.4% vs. 23.5% (P<0.05), respectively. p120ctn expression was positively correlated with E-cadherin expression (r=0.404, P<0.05), but negatively with nuclear NF-κB expression (r= - 0.347, P<0.05). Western blotting showed that as compared with the normal controls, the p120ctn protein level was significantly decreased (P<0.05), whereas the NF-κB protein level was increased (P<0.05) in UC tissues. It was concluded that in the colonic tissues of UC patients, the expressions of E-cadherin, p120ctn and β-catenin are decreased, suggesting the mucosal barrier is impaired in UC. Moreover, NF-κB is increased and activated in the UC tissues, resulting in the inflammation in UC. p120ctn may influence the UC development through modulating intercellular adhesion and inflammatory response.


Subject(s)
Adolescent , Adult , Cadherins , Metabolism , Catenins , Metabolism , Colitis, Ulcerative , Metabolism , Pathology , Down-Regulation , Female , Humans , Male , Middle Aged , NF-kappa B , Metabolism , Statistics, Nonparametric , Young Adult , beta Catenin , Metabolism
7.
Chinese Medical Journal ; (24): 2918-2922, 2013.
Article in English | WPRIM | ID: wpr-263557

ABSTRACT

<p><b>BACKGROUND</b>p120 catenin (p120ctn) is an adheren junction protein that regulates barrier function, but its role has not been explored in alveolar edema induced by ventilation. We measured stretch-induced cell gap formation in MLE 12 cells due to the loss of p120. We hypothesized that alveolar permeability was increased by high lung inflation associated with alveolar epithelia cell tight junctions being destroyed, which resulted from the loss of p120.</p><p><b>METHODS</b>Cultured MLE12 cells were subjected to being stretched or un-stretched (control) and some cells were pretreated with pp2 (c-src inhibitor). After the end of stretching for 0, 1, 2, and 4 hours, the cells were lysed, and p120 expression and c-src activation was determined by Western blotting analysis. In vivo, SD rats were taken to different tidal volumes (Vt 7 ml/kg or 40 ml/kg, PEEP = 0, respiratory rate 30-40 betas/min) for 0, 1, 2, and 4 hour and some were pretreated with pp2, and alveolar edema was calculated.</p><p><b>RESULTS</b>It was found that p120 expression was reduced and c-src activation increased in a time-dependent and strain-dependent manner due to cyclic-stretch of the alveolar epithelial cells. These changes could be reversed by inhibition of c-src. We obtained similar changes in rats when they were subjected to large tidal volumes and the alveolar edema increased more than in rats in the low Vt group. Pretreated the rats with inhibition of c-src had less pulmonary edema induced by the high tidal volume ventilation.</p><p><b>CONCLUSIONS</b>Cyclic stretch MLE 12 cells induced the loss of p120 and may be the same reason by high tidal volume ventilation in rats can aggravate alveolar edema. Maintenance of p120 expression may be a novel therapeutic strategy for the prevention and treatment of ventilation induced lung injury (VILI).</p>


Subject(s)
Animals , Blotting, Western , Catenins , Physiology , Cells, Cultured , Mice , Pulmonary Alveoli , Pathology , Pulmonary Edema , Pathology , Rats , Rats, Sprague-Dawley , Tidal Volume , Ventilator-Induced Lung Injury , Pathology
8.
Chinese Medical Journal ; (24): 3057-3063, 2013.
Article in English | WPRIM | ID: wpr-263525

ABSTRACT

<p><b>BACKGROUND</b>Heme oxygenase-1 (HO-1) can be induced by inflammatory cytokines, oxidation, ischemia, hypoxia, and endotoxins. As a "graft survival protective gene," HO-1 is a hot spot in organ transplantation research. However, the role of HO-1 gene expression in the function of human colon adenocarcinoma cell line (Caco-2) cells has not been reported previously.</p><p><b>METHODS</b>The role of HO-1 in the proliferation and migration of Caco-2 cells was analyzed using a stable HO-1 expression plasmid. We constructed a recombinant adeno-associated virus plasmid containing the HO-1 gene, heme oxygenase 1 (HMOX1), which was transfected into Caco-2 intestinal cells. We identified a number of target genes by global microarray analysis combined with real-time polymerase chain reaction (PCR) and chromatin immunoprecipitation assay.</p><p><b>RESULTS</b>Our results showed that significant HO-1 upregulation was demonstrated in the Caco-2 cells after HO-1 transfection. Restoration of HO-1 expression promoted proliferation and invasion in vitro. The CTNND1 gene, a member of the armadillo protein family, was identified as a direct HO-1 target gene.</p><p><b>CONCLUSION</b>Overexpression of HO-1 promotes Caco-2 cell proliferation and migration by targeting the CTNND1 gene.</p>


Subject(s)
Caco-2 Cells , Catenins , Genetics , Cell Movement , Cell Proliferation , Gene Expression Regulation, Neoplastic , Heme Oxygenase-1 , Genetics , Physiology , Humans , Real-Time Polymerase Chain Reaction
9.
Chinese Journal of Pathology ; (12): 599-603, 2013.
Article in Chinese | WPRIM | ID: wpr-233384

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the clinicopathologic features, clinical progress and prognosis of the basal-like subtype of invasive lobular carcinoma (ILC) of the breast.</p><p><b>METHODS</b>Four cases of ILC were analyzed by detailed histopathologic observation and immunohistochemical staining for E-cadherin, p120 catenin, ER, PR, HER2, CK5/6, EGFR, p63, p53, Ki-67 using MaxVision method. The follow-up and clinical data were analyzed.</p><p><b>RESULTS</b>Morphologically, one case was mixed ILC and three cases were pleomorphic ILC. The tumor cells were negative for E-cadherin except one case with focal membrane positivity, and all showed p120 catenin cytoplasmic positivity except one case with focal membrane positivity. All cases were negative for ER, PR and HER2 (triple negative), and positive for EGFR and CK5/6. Two cases were positive for p63. The cases were partly and weakly positive for p53, and the Ki-67 positive rate was between 30% and 75%. Follow-up data showed that two cases developed chest wall metastases, and in one case, there was progression to liver and abdominal metastases.</p><p><b>CONCLUSIONS</b>ILC of the breast are ER, PR and HER2 "triple negative", CK5/6 and EGFR positive, indicative of basal-like characteristics. Basal-like subtype of ILC are peculiarly prone to metastasis and poor response to chemotherapy, suggesting that it is associated with poor prognosis.</p>


Subject(s)
Adult , Aged , Antineoplastic Combined Chemotherapy Protocols , Therapeutic Uses , Biomarkers, Tumor , Metabolism , Breast Neoplasms , Drug Therapy , Metabolism , Pathology , General Surgery , Cadherins , Metabolism , Carcinoma, Lobular , Drug Therapy , Metabolism , Pathology , General Surgery , Catenins , Metabolism , Combined Modality Therapy , Female , Follow-Up Studies , Humans , Immunohistochemistry , Keratin-5 , Metabolism , Keratin-6 , Metabolism , Ki-67 Antigen , Metabolism , Liver Neoplasms , Lymphatic Metastasis , Mastectomy, Modified Radical , Middle Aged , ErbB Receptors , Metabolism , Receptor, ErbB-2 , Metabolism , Receptors, Estrogen , Metabolism , Receptors, Progesterone , Metabolism , Thoracic Neoplasms , Thoracic Wall , Tumor Suppressor Protein p53 , Metabolism
10.
Int. braz. j. urol ; 38(4): 466-473, July-Aug. 2012. ilus, tab
Article in English | LILACS | ID: lil-649439

ABSTRACT

INTRODUCTION: Cell adhesion molecules (CAM) are required for maintaining a normal epithelial phenotype, and abnormalities in CAM expression have been related to cancer progression, including bladder urothelial carcinomas. There is only one study that correlates E-cadherin and α-, β- and γ-catenin expression with prognosis of upper tract urothelial carcinomas. Our aim is to study the pattern of immune expression of these CAMs in urothelial carcinomas from the renal pelvis and ureter in patients who have been treated surgically. Our goal is to correlate these expression levels and characteristics with well-known prognostic parameters for disease-free survival. MATERIALS AND METHODS: We evaluated specimens from 20 patients with urothelial carcinomas of the renal pelvis and ureter who were treated with nephroureterectomy or ureterectomy between June 1997 and January 2007. CAM expression was evaluated by immunohistochemistry in a tissue microarray and correlated with histopathological characteristics and patient outcomes after a mean follow-up of 55 months. RESULTS: We observed a relationship between E-cadherin expression and disease recurrence. Disease recurrence occurred in 87.5% of patients with strong E-cadherin expression. Only 50.0% of patients with moderate expression and 0% of patients with weak or no expression of E-cadherin had disease recurrence (p = 0.014). There was also a difference in disease-free survival. Patients with strong E-cadherin expression had a mean disease-free survival rate of 49.1 months, compared to 83.9 months for patients with moderate expression (p = 0.011). Additionally, an absence of α-catenin expression was associated with tumors that were larger than 3 cm (p = 0.003). CONCLUSIONS: We demonstrated for the first time that immune expression of E-cadherin is related to tumor recurrence and disease-free survival rates, and the absence of α-catenin expression is related to tumor size in upper tract urothelial carcinomas.


Subject(s)
Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Cadherins/analysis , Carcinoma/chemistry , Catenins/analysis , Biomarkers, Tumor/analysis , Ureteral Neoplasms/chemistry , Urinary Tract/chemistry , Carcinoma/pathology , Cell Adhesion Molecules/analysis , Epidemiologic Methods , Immunohistochemistry , Prognosis , Sex Distribution , Time Factors , Tissue Array Analysis , Ureteral Neoplasms/pathology , Urinary Tract/pathology , alpha Catenin/analysis , beta Catenin/analysis , gamma Catenin/analysis
11.
Chinese Journal of Pathology ; (12): 613-617, 2012.
Article in Chinese | WPRIM | ID: wpr-303508

ABSTRACT

<p><b>OBJECTIVE</b>To study the clinicopathologic features of pure mucinous carcinomas of the breast with diffuse micropapillary pattern.</p><p><b>METHODS</b>Twenty-six cases of micropapillary variant of pure mucinous carcinoma of the breast were retrospectively reviewed by light microscopy, immunohistochemistry and clinical data analyses.</p><p><b>RESULTS</b>The age of 26 female patients ranged from 30 to 77 years old, of which 12 cases with clinical details available were mean 54 years old. The tumor diameter ranged from 0.8 to 9.0 cm (mean 3.2 cm). Ipsilateral axillary nodal metastases were identified in 3 cases. Cutaneous involvement was also found in 2 cases. The tumor cells showed the similar architectural arrangement as in invasive micropapillary carcinoma, with peripheral borders of the cell clusters highlighted by epithelial membrane antigen. Various amount of mucin occupied the retraction spaces around the tumor cells. Compared with conventional pure mucinous carcinoma of the breast, mucinous carcinomas with micropapillary pattern showed different nuclear grades (19 cases of grade I, 2 cases of grade II, 5 cases of grade III). The micropapillary cell clusters varied in size (22 cases of big micropapillary and 4 cases of small). Intraductal carcinoma was observed in 12 cases. Calcification and psammoma bodies were observed in 8 cases. Immunophenotyping, the tumor cells were with higher expression of hormone receptors, but HER2 were negative. Ki-67 positive index was 1% ∼ 70%. Neuroendocrine differentiation was observed in 6 cases.</p><p><b>CONCLUSIONS</b>The micropapillary variant of pure mucinous carcinoma of the breast, which mainly occurs in younger women, may carry the similar propensity for angioinvasion and nodal metastasis as infiltrating micropapillary carcinoma at least in cases with high nuclear grade. This morphologic subtype needs to be distinguished from conventional pure mucinous carcinoma of the breast and treated properly.</p>


Subject(s)
Adenocarcinoma, Mucinous , Metabolism , Pathology , General Surgery , Adult , Aged , Breast Neoplasms , Metabolism , Pathology , General Surgery , Cadherins , Metabolism , Carcinoma, Papillary , Metabolism , Pathology , General Surgery , Catenins , Metabolism , Female , Follow-Up Studies , Humans , Immunohistochemistry , Ki-67 Antigen , Metabolism , Lymph Node Excision , Lymphatic Metastasis , Mastectomy, Modified Radical , Methods , Middle Aged , Mucin-1 , Metabolism , Receptors, Estrogen , Metabolism , Receptors, Progesterone , Metabolism , Retrospective Studies
12.
Article in English | WPRIM | ID: wpr-233095

ABSTRACT

This study examined the possible role of p120ctn in the pathogenesis and development of pancreatic cancer. PANC-1 cells, a kind of human pancreatic carcinoma cell line, were cultured in this study. p120ctn was immunocytochemically detected in PANC-1 cells. The recombinant lentivirus vector was constructed to knock down the p120ctn expression of PANC-1 cells. Real-time quantitative PCR (RQ-PCR) and Western blotting were used to determine the expression of p120ctn and E-cadherin in PANC-1 cells after p120ctn knockdown. The adhesion, invasion and migration capacity of PANC-1 cells after p120ctn knockdown was detected by cell adhesion, invasion and migration assays. Cell growth was measured by the MTT method. Cell cycle and apoptosis were analyzed by fluorescence-activated cell sorting. The results showed that p120ctn knockdown led to significantly down-regulated E-cadherin and a reduced cell-to-cell adhesion ability in PANC-1 cells. shRNA-mediated knockdown of p120ctn reduced invasion and migration capacity of PANC-1 cells, inhibited cell growth, caused a significant decrease in the percentage of cells in G(1), an increase in S, and promoted apoptosis of PANC-1 cells. It was concluded that p120ctn plays a pivotal role in the proliferation and metastasis of pancreatic carcinoma, suggesting that p120ctn is a novel target for pancreatic carcinoma treatment.


Subject(s)
Catenins , Genetics , Cell Line, Tumor , Gene Silencing , Humans , Pancreatic Neoplasms , Genetics
13.
Chinese Journal of Pathology ; (12): 741-744, 2011.
Article in Chinese | WPRIM | ID: wpr-358248

ABSTRACT

<p><b>OBJECTIVE</b>To investigate expressions of E-cadherin (E-cad), p120catenin (p120), 34βE12 in invasive lobular carcinomas of the breast and their roles of diagnoses.</p><p><b>METHODS</b>The 81 cases of ILC, including 67 cases of pure type and 14 cases of ductal-lobular mixed type, which had been diagnosed in our department were collected and immunohistochemistry of E-cad, p120 and 34βE12 were performed. All the cases were diagnosed again according to morphology and immunophenotypes (MaxVision method), and difference of diagnoses and expressions of the three indexes were analysed.</p><p><b>RESULTS</b>Sixty four of 81 cases were permantly diagnosed of ILC. In the 61 cases of pure type, 54 cases displayed E-cad negative and p120 cytoplastic positive, 1 case displayed E-cad negative and p120 atypical positive, 3 cases displayed E-cad membrane positive and p120 cytoplastic positive, and 3 cases displayed both atypical positive. Fifty two of 61 cases displayed 34βE12 positive. The 3 cases of mixed type displayed p120 cytoplastic positive, and 2 cases displayed E-cad negative and 1 case displayed atypical positive. All the 3 cases displayed 34βE12 positive.</p><p><b>CONCLUSIONS</b>The diagnosis of ILC is one of the most difficult problems in breast pathology, and combination of E-cad and p120 immunostaining is an effective method for assistance. It needs further studies for invasive ductal carcinomas with morphological features of lobular carcinomas.</p>


Subject(s)
Adult , Aged , Breast Neoplasms , Diagnosis , Metabolism , Pathology , Cadherins , Metabolism , Carcinoma, Ductal, Breast , Diagnosis , Metabolism , Pathology , Carcinoma, Lobular , Diagnosis , Metabolism , Pathology , Catenins , Metabolism , Diagnosis, Differential , Female , Humans , Immunohistochemistry , Keratins , Metabolism , Middle Aged , Retrospective Studies
14.
Chinese Journal of Hepatology ; (12): 599-602, 2011.
Article in Chinese | WPRIM | ID: wpr-330686

ABSTRACT

To investigate the relationship and significance of Wnt/b-catenin signaling pathway with caspase-3, XIAP, HSP27and Grp-78. The HCC cell line HepG2 was transfected with small interfering RNA (siRNA) directed against b-catenin. After 72 and 96 h, protein was extracted and the protein expressions of b-catenin, caspase-3, XIAP, Grp-78 and HSP27 were detected by Western blot. b-catenin protein expression was inhibited at both time points and the expression at 96 h was higher than that at 72 h (F = 160.72, P is less than to 0.01). Interestingly, Caspase-3 protein expression was decreased at 72 h and increased to normal at 96 h (F = 136.10, P is less than to 0.01), while p-caspase-3 protein expression increased at 72 h and decreased to normal at 96 h (F = 98.65, P is less than to 0.01). XIAP protein expression decreased at 72 h (F = 37.29, P is less than to 0.01) and increased at 96 h. Grp-78 protein expression increased at 72 h and decreased to normal at 96 h ( F = 58.72, P is less than to 0.01). HSP27 protein expression showed no change following transfection ( F = 1.91, P is more than to 0.05). Wnt/b-catenin signaling pathway is related to the protein expressions of caspase-3, XIAP and Grp-78, but not related to HSP27 protein expression in HCC. Wnt/b-catenin signaling pathway may participate in the regulation of HCC apoptosis, proliferation and differentiation through affecting these factors.


Subject(s)
Carcinoma, Hepatocellular , Caspase 3 , Catenins , Humans , Liver Neoplasms , Wnt Signaling Pathway , beta Catenin , Metabolism
15.
Article in Chinese | WPRIM | ID: wpr-301596

ABSTRACT

<p><b>OBJECTIVE</b>To explore the effects of K-ras gene mutation on colon cancer cell line Caco-2 metastasis by regulating E-cadherin/beta-catenin/p120 protein complex formation and RhoA protein activity.</p><p><b>METHODS</b>K-ras wild-type colon cancer cell line Caco-2 was transiently transfected by phr-GFP vector (control group), transfected by mutant K-ras gene phr-K-ras (Val12) vector (transfection group), transfected by mutant K-ras gene phr-K-ras (Val12) vector and treated by specific MAPK pathway inhibitor PD98059 (MAPK inhibition group), or transfected by mutant K-ras gene phr-K-ras (Val12) vector and treated by specific PI-3K pathway inhibitor LY294002 (PI-3K inhibition group), respectively. Cell migration was tested by Transwell experiment. E-cadherin and beta-catenin protein expression and intracellular location were detected by cell immunofluorescence method. Intracellular p120 protein expression was detected by Western blot. beta-catenin protein level which combined with E-cadherin was detected by immunoprecipitation. RhoA activity was analyzed by Pull-down assay.</p><p><b>RESULTS</b>The Caco-2 cell migration rate was (19.8 +/- 5.6) % in transfection group, which was significantly higher than that in control group [(14.0 +/- 4.2) %] (P = 0.001) and in MAPK inhibition group [(15.8 +/- 1.2) %] (P = 0.044), but was not significantly different from that in PI-3K inhibition group [(17.5 +/- 2.8) %] (P = 0.095). Immunofluorescence method showed that the E-cadherin and beta-catenin stain located in the cell membrane decreased in transfection group. Western blot showed that the total intracellular p120 protein decreased in transfection group and PI-3K inhibition group. Immunoprecipitation data showed that beta-catenin protein level combined with E-cadherin decreased in transfection group and PI-3K group. Pull-down test showed that RhoA protein activity was up-regulated in transfection group.</p><p><b>CONCLUSION</b>K-ras gene mutation stimulates the migration of colon cancer cell Caco-2, which may be achieved by decreasing the E-cadherin/beta-catenin/p120 protein complex formation via MAPK pathway and increasing the RhoA protein activity.</p>


Subject(s)
Caco-2 Cells , Cadherins , Metabolism , Catenins , Metabolism , Cell Movement , Colonic Neoplasms , Metabolism , Pathology , Genes, ras , Genetics , Humans , Multiprotein Complexes , Metabolism , Mutation , Neoplasm Metastasis , Transfection , beta Catenin , Metabolism , rhoA GTP-Binding Protein , Metabolism
16.
Chinese Journal of Oncology ; (12): 273-277, 2010.
Article in Chinese | WPRIM | ID: wpr-260419

ABSTRACT

<p><b>OBJECTIVE</b>To determine how patients with infiltrating lobular carcinoma (ILC) differ from patients with the more common infiltrating ductal carcinoma (IDC), and observe the different expression patterns of E-cadherin and p120-catenin proteins in both ILCs and IDCs.</p><p><b>METHODS</b>The patients with ILC admitted to our hospital from Jan 1999 to Dec 2006 and patients with IDC from Jan 2000 to Dec 2000 were included in this study. All their pathological slides were reviewed, and their clinical data and treatment variables were analyzed retrospectively. Then the expression patterns of E-cadherin and p120-catenin proteins in both ILCs and IDCs were detected by immunohistochemistry on tissue microarray.</p><p><b>RESULTS</b>The 5-year overall survival was 81.7% for ILCs and 79.1% for IDCs (P = 0.055). The 5-year disease-free survival was 61.8% for ILCs and 83.7% for IDCs (P < 0.001). Cytoplasmic localization of p120-catenin and loss of E-cadherin expression were more common in ILCs than in IDCs. The complete losses of E-cadherin in ILCs and IDCs were 55.6% (20/36) and 20.4% (45/221, P < 0.001), respectively. The p120-catenin showed a diffuse cytoplasmic localization in 66.7% (24/36) of ILCs and 16.3% (36/221) of IDCs (P < 0.001). Interestingly, the cytoplasmic localization of p120-catenin was clearly associated with the absence of E-cadherin expression in ILCs (P = 0.002), cytoplasmic localization of p120-catenin and absence of E-cadherin expression were observed 55.6% (20/36) in ILCs compared with 4.1% (9/221) in IDCs (P < 0.001).</p><p><b>CONCLUSION</b>ILC has several specific biological and prognostic characteristics which are different in IDC. Different expression patterns of E-cadherin and p120-catenin proteins can be helpful to recognize ILC from IDC.</p>


Subject(s)
Bone Neoplasms , Breast Neoplasms , Metabolism , Pathology , Cadherins , Metabolism , Carcinoma, Ductal, Breast , Metabolism , Pathology , Carcinoma, Lobular , Metabolism , Pathology , Catenins , Metabolism , Cytoplasm , Metabolism , Diagnosis, Differential , Disease-Free Survival , Female , Follow-Up Studies , Gene Expression Regulation, Neoplastic , Humans , Lung Neoplasms , Male , Middle Aged , Neoplasm Recurrence, Local , Retrospective Studies , Survival Rate
17.
São Paulo; s.n; 2009. [122] p. ilus, tab, graf.
Thesis in Portuguese | LILACS | ID: lil-587008

ABSTRACT

Objetivo: As moléculas de adesão celular (MAC) são essenciais para a manutenção do fenótipo epitelial. Alguns estudos têm relatado associação entre as alterações de sua expressão e a carcinogênese, mas o seu papel no câncer de próstata não é claro. Nosso objetivo foi estudar o perfil de expressão de E-caderina, cateninas e integrinas em espécimes cirúrgicos de câncer de próstata e associar as suas expressões com a evolução do tumor. Avaliamos também o perfil de expressão em metástases ósseas e linfonodais, a fim de compreender a influência destes marcadores na progressão do câncer de próstata. Materiais e Métodos: Foram selecionados 111 pacientes com câncer de próstata localizado tratados com prostatectomia radical pelo mesmo cirurgião. Sessenta pacientes não apresentaram recidiva tumoral após acompanhamento médio de 123 meses. A expressão das MAC foi avaliada por imuno-histoquímica (IH) em microarranjo tecidual (TMA), contendo duas amostras de cada tumor. Empregamos análise semiquantitativa para avaliação da expressão e determinamos a associação entre a expressão de cada MAC com a recorrência do tumor após a cirurgia. Avaliamos também a expressão das MAC por IH em TMA contendo espécimes de 28 metástases ósseas e em outro TMA contendo 19 metástases linfonodais com seus 19 tumores primários correspondentes. Resultados: Nos tumores primários a análise multivariada mostrou que a expressão das integrinas 3 e 3 1 relaciona-se com recidiva da doença. Quando a expressão de 3 foi forte e a expressão de 3 1 foi positiva, as chances de recorrência foram de 3,0 e 2,5 vezes maior. Apenas 19% e 28% dos pacientes estavam livres de recidiva após seguimento médio de 123 meses, quando os tumores apresentavam forte imunoexpressão de 3 ou positiva para 3 1 respectivamente. Outras integrinas apresentaram expressão reduzida, exceto 6 que foi expressa pela maioria dos tumores primário e metástases. A E-Caderina e as cateninas não mostraram associação com o prognóstico no tumor...


Purpose: Cell adhesion molecules (CAM) are essential for the maintenance of epithelial phenotype. Some studies have reported correlations between abnormalities in their expression and carcinogenesis, but their role in prostate cancer is unclear. Our aim was to study the expression profile of E-cadherin, catenins and integrins in surgical specimens of prostate cancer and associate their expression with outcome. We also assessed these expressions in bone and lymph node metastases in order to understand their influence in the progression of prostate cancer. Materials and Methods: We selected 111 patients with localized prostate cancer who underwent radical prostatectomy performed by the same surgeon. Sixty patients had no tumor recurrence after a median follow-up of 123 months. The CAM expression was evaluated by immunohistochemistry in a tissue microarray (TMA) containing two samples of each tumor. A semiquantitative analysis was employed and we measured the association between the expression of CAM and tumor recurrence. We also evaluated CAM expression by immunohistochemistry in a TMA containing 28 bone metastases and in other TMA containing 19 lymph node metastases with their corresponding 19 primary tumors. Results: In primary tumors, multivariate analysis showed that expression of 3 and 31 integrins was related to worse outcome. When 3 expression was strong and 31 expression was positive,the odds of recurrence were 3.0 and 2.5 fold higher. Only 19% and 28% of patients were recurrence-free in a mean follow up period of 123 months, when tumors showed strong 3 or positive 31 immuno-expression respectively. Other integrins have shown reduced expression, except 6 , which was expressed in most primary and metastatic cases. E-cadherin and catenins expressions were not associated with primary tumor outcome. At the metastatic setting, there was a global loss of CAM expression. We observed reliable gain of expression with prostate cancer progression...


Subject(s)
Humans , Male , Adult , Middle Aged , Cadherins , Catenins , Cell Adhesion Molecules , Integrins , Prostatic Neoplasms
18.
Article in Chinese | WPRIM | ID: wpr-748672

ABSTRACT

OBJECTIVE@#To study the expression of E-cadherin and P(120ctn) in nasopharyngeal carcinoma tissues, and to investigate their relationship and the relation with clinico-pathological features.@*METHOD@#Two-step immunohistochemical staining was applied to detect the expression of E-cadherin and P(120ctn) in formalin fixation and paraffin-embedded specimens from 56 cases with nasopharyngeal carcinoma and 15 cases with normal nasopharyngeal epithelia.@*RESULT@#The abnormal expression rates of E-cadherin and P(120ctn) in the 56 cases of NPC tissues were 64.29% and 67.86% respectively, mainly with reduction of expression membrane and with the expression of cytoplasm; 6.67% of the 15 comparative normal cases of nasopharyngitis had abnormal expression of E-cadherin and P(120ctn). The differences were statistically significant. The abnormal expression rates of E-cadherin and P(120ctn) in NPC tissues were 71.43% and 85.71% respectively in low differentiated cancer group, which was obviously higher than the rates-42.86% and 36.29%-in high and middle differentiated cancer group. The 80.00% and 85.00% abnormal expression rate in the group with cervical lymph node metastases was higher than that in the group without cervical lymph node metastases (52.78%, 58.33%). The abnormal expression rate of E-cadherin and P(120ctn) (76.92%, 84.62%) in the third and forth phases was higher than that in the first and second phases (46.66%, 53.33%). The differences were statistically significant (P < 0.05). There were all together 12 co-expression cases of P120ctn) and E-cadherin and 28 abnormal co-expression cases in the 56 cases of NPC tissues, which was of obvious consistency and correlation, with the relevant indexes: rs = 0.5217 and P < 0.01.@*CONCLUSION@#The abnormal expression of E-cadherin and P(120ctn) is closely related to the degree of differentiation, clinical stage and cervical lymph node metastasis, and they join in the process of NPC initiation, progression, invasion and metastasis.


Subject(s)
Adult , Aged , Cadherins , Metabolism , Catenins , Metabolism , Female , Humans , Male , Middle Aged , Nasopharyngeal Neoplasms , Metabolism , Pathology , Neoplasm Staging , Young Adult
19.
Chinese Journal of Surgery ; (12): 1809-1812, 2009.
Article in Chinese | WPRIM | ID: wpr-290990

ABSTRACT

<p><b>OBJECTIVES</b>To investigate the expression of P120 catenin in pancreatic carcinoma and to explore the association between P120 catenin gene polymorphism at T755G position and pancreatic carcinoma.</p><p><b>METHODS</b>The expression of P120 catenin in 52 cases of pancreatic carcinoma and normal pancreatic tissues on the mRNA and protein levels were evaluated by RT-PCR and Western Blot methods respectively. P120 catenin gene polymorphism at T755G position of in 52 patients and 60 healthy controls were examined by PCR-restriction fragment length polymorphism (PCR-RFLP) technique.</p><p><b>RESULTS</b>The mRNA and protein expressions of P120 catenin in pancreatic carcinoma tissues were significantly lower than normal pancreatic tissues (P=0.000, P=0.002). Reduced expression of P120 catenin mRNA was significantly correlated with differentiated (P=0.033), lymph node metastasis (P=0.004), vascular invasion (P=0.022), and pTNM stage (P=0.003). Additionally, there were significant difference of P120 catenin gene polymorphism genotypes and alleles at T755G position between patients and healthy controls (P=0.008, P=0.016). The GG genotype of P120 catenin gene was associated with higher risk of incidence for pancreatic carcinoma compared with the TT genotype (OR=2.765, 95%CI=1.312-3.958).</p><p><b>CONCLUSIONS</b>The reduced expressions of both P120 catenin mRNA and protein in pancreatic carcinoma suggest its association with pancreatic carcinoma development. Polymorphism of P120 catenin gene at T755G situation might be a risk factor for pancreatic carcinoma, and it may be used to diagnosis and prevent pancreatic carcinoma early.</p>


Subject(s)
Case-Control Studies , Catenins , Genetics , Metabolism , Female , Humans , Male , Middle Aged , Pancreatic Neoplasms , Genetics , Metabolism , Polymorphism, Genetic
20.
Chinese Journal of Pathology ; (12): 663-667, 2009.
Article in Chinese | WPRIM | ID: wpr-249071

ABSTRACT

<p><b>OBJECTIVE</b>To study the diagnostic usefulness of immunohistochemical markers in distinguishing between invasive ductal carcinoma and invasive lobular carcinoma of breast.</p><p><b>METHODS</b>Twenty-four cases of grade I invasive ductal carcinoma, 12 cases of classic invasive lobular carcinoma and 14 cases of invasive carcinoma with mixed ductal and lobular features were retrieved from the archival files of Peking University First Hospital during the period from January, 1998 to December, 2001. Immunohistochemical study for E-cadherin, p120 catenin, epithelia membrane protein 1 (EMP1) and DVL1 was performed.</p><p><b>RESULTS</b>The positivity rates for E-cadherin in grade I invasive ductal carcinoma and classic invasive lobular carcinoma were 83.3% (20/24) and 0, respectively (P < 0.01). The positivity rates for p120 catenin were 100% in both grade 1 invasive ductal carcinoma (membranous staining) and classic invasive lobular carcinoma (cytoplasmic staining). The positivity rates for EMP1 and DVL1 in gradeI invasive ductal carcinoma were 95.8% (23/24) and 54.2% (13/24), respectively; while those in classic invasive lobular carcinoma were 12 and 5 cases, respectively.</p><p><b>CONCLUSIONS</b>E-cadherin and p120 catenin are useful immunomarkers for distinguishing between invasive ductal carcinoma and invasive lobular carcinoma. On the other hand, EMP1 and DVL1 are of limited value in this respect.</p>


Subject(s)
Adaptor Proteins, Signal Transducing , Metabolism , Breast Neoplasms , Metabolism , Pathology , General Surgery , Cadherins , Metabolism , Carcinoma, Ductal, Breast , Metabolism , Pathology , General Surgery , Carcinoma, Lobular , Metabolism , Pathology , General Surgery , Catenins , Metabolism , Diagnosis, Differential , Disease-Free Survival , Dishevelled Proteins , Female , Humans , Mastectomy , Methods , Middle Aged , Neoplasm Proteins , Metabolism , Phosphoproteins , Metabolism , Receptors, Cell Surface , Metabolism , Survival Rate
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