This paper aimed to investigate the effect of total flavonoids of buckwheat flower and leaf on myocardial cell apoptosis and Wnt/β-catenin/peroxisome proliferator-activated receptor γ(PPARγ) pathway in arrhythmic rats. SD rats were randomly divided into a control group, a model group, a low-dose(20 mg·kg~(-1)) group of total flavonoids of buckwheat flower and leaf, a medium-dose(40 mg·kg~(-1)) group of total flavonoids of buckwheat flower and leaf, a high-dose(80 mg·kg~(-1)) group of total flavonoids of buckwheat flower and leaf, a propranolol hydrochloride(2 mg·kg~(-1)) group, with 12 rats in each group. Except the control group, rats in other groups were prepared as models of arrhythmia by sublingual injection of 1 mL·kg~(-1) of 0.002% aconitine. After grouping and intervention with drugs, the arrhythmia, myocardial cells apoptosis, myocardial tissue glutathione peroxidase(GSH-Px), catalase(CAT), malondialdehyde(MDA), serum interleukin-6(IL-6), prostaglandin E2(PGE2) levels, myocardial tissue apoptosis, and Wnt/β-catenin/PPARγ pathway-related protein expression of rats in each group were measured. As compared with the control group, the arrhythmia score, the number of ventricular premature beats, ventricular fibrillation duration, myocardial cell apoptosis rate, MDA levels in myocardial tissues, serum IL-6 and PGE2 levels, Bax in myocardial tissues, and Wnt1 and β-catenin protein expression levels increased significantly in the model group, whereas the GSH-Px and CAT levels, and Bcl-2 and PPARγ protein expression levels in myocardial tissues reduced significantly. As compared with the model group, the arrhythmia score, the number of ventricular premature beats, ventricular fibrillation duration, myocardial cell apoptosis rate, MDA leve in myocardial tissues, serum IL-6 and PGE2 levels, Bax in myocardial tissues, and Wnt1 and β-catenin protein expression levels reduced in the drug intervention groups, whereas the GSH-Px and CAT levels and Bcl-2 and PPARγ protein expression levels in myocardial tissues increased. The groups of total flavonoids of buckwheat flower and leaf were in a dose-dependent manner. There was no significant difference in the levels of each index in rats between the propranolol hydrochloride group and the high-dose group of total flavonoids of buckwheat flower and leaf. The total flavonoids of buckwheat flower and leaf inhibit the activation of Wnt/β-catenin pathway, up-regulate the expression of PPARγ, reduce oxidative stress and inflammatory damage in myocardial tissues of arrhythmic rats, reduce myocardial cell apoptosis, and improve the symptoms of arrhythmia in rats.
Rats ; Animals ; PPAR gamma/metabolism* ; Fagopyrum/genetics* ; Rats, Sprague-Dawley ; bcl-2-Associated X Protein ; beta Catenin/metabolism* ; Interleukin-6 ; Flavonoids/pharmacology* ; Propranolol/pharmacology* ; Ventricular Fibrillation ; Dinoprostone ; Wnt Signaling Pathway ; Plant Leaves/metabolism* ; Flowers/metabolism* ; Apoptosis ; Cardiac Complexes, Premature

Objective: To investigate the relationship between the expression levels of Plakoglobin protein in residual lesions after neoadjuvant chemotherapy (NAC) and the prognosis of breast cancer patients. Methods: Clinical and pathological data from 174 breast cancer patients who underwent surgery after receiving NAC at the Cancer Hospital of Chinese Academy of Medical Sciences from January 2009 to December 2017 were collected. The expression level of Plakoglobin in residual cancer lesions was evaluated by immunohistochemistry. The correlation between Plakoglobin expression level and clinicopathological features was analyzed. Survival analysis was performed using the Kaplan-Meier method, and Cox proportional hazard regression models were used for factor analysis. Results: Among the 174 patients, 140 had low expression of Plakoglobin, and 34 had high expression. The median disease-free survival (DFS) and overall survival (OS) in the Plakoglobin low expression group were 59.46 and 71.68 months, respectively, both of which were higher than those in the high expression group (36.58 and 47.26 months, respectively, both P<0.05). Univariate analysis showed that Plakoglobin expression, pathological N stage, lymphovascular invasion status, histological grade, Ki-67, and molecular subtypes were associated with OS (all P<0.05), while pathological N stage, histological grade, and Ki-67 were associated with DFS (all P<0.05). Multivariate analysis revealed that Plakoglobin expression (HR=2.438, 95% CI: 1.256-4.735, P=0.008) was an independent predictor for OS, and Ki-67 (HR=2.228, 95% CI: 1.316-3.773, P=0.003) was an independent predictor for DFS. Conclusion: In breast cancer patients with residual lesions after NAC, those with low Plakoglobin expression have relatively longer OS and Plakoglobin is an independent prognostic factor for OS.
Humans ; Female ; Prognosis ; Breast Neoplasms/surgery* ; Ki-67 Antigen/analysis* ; Neoadjuvant Therapy/methods* ; gamma Catenin ; Neoplasm, Residual ; Disease-Free Survival ; Retrospective Studies ; Antineoplastic Combined Chemotherapy Protocols/therapeutic use*

Objective: To investigate the relationship between the expression levels of Plakoglobin protein in residual lesions after neoadjuvant chemotherapy (NAC) and the prognosis of breast cancer patients. Methods: Clinical and pathological data from 174 breast cancer patients who underwent surgery after receiving NAC at the Cancer Hospital of Chinese Academy of Medical Sciences from January 2009 to December 2017 were collected. The expression level of Plakoglobin in residual cancer lesions was evaluated by immunohistochemistry. The correlation between Plakoglobin expression level and clinicopathological features was analyzed. Survival analysis was performed using the Kaplan-Meier method, and Cox proportional hazard regression models were used for factor analysis. Results: Among the 174 patients, 140 had low expression of Plakoglobin, and 34 had high expression. The median disease-free survival (DFS) and overall survival (OS) in the Plakoglobin low expression group were 59.46 and 71.68 months, respectively, both of which were higher than those in the high expression group (36.58 and 47.26 months, respectively, both P<0.05). Univariate analysis showed that Plakoglobin expression, pathological N stage, lymphovascular invasion status, histological grade, Ki-67, and molecular subtypes were associated with OS (all P<0.05), while pathological N stage, histological grade, and Ki-67 were associated with DFS (all P<0.05). Multivariate analysis revealed that Plakoglobin expression (HR=2.438, 95% CI: 1.256-4.735, P=0.008) was an independent predictor for OS, and Ki-67 (HR=2.228, 95% CI: 1.316-3.773, P=0.003) was an independent predictor for DFS. Conclusion: In breast cancer patients with residual lesions after NAC, those with low Plakoglobin expression have relatively longer OS and Plakoglobin is an independent prognostic factor for OS.
Humans ; Female ; Prognosis ; Breast Neoplasms/surgery* ; Ki-67 Antigen/analysis* ; Neoadjuvant Therapy/methods* ; gamma Catenin ; Neoplasm, Residual ; Disease-Free Survival ; Retrospective Studies ; Antineoplastic Combined Chemotherapy Protocols/therapeutic use*

OBJECTIVETo investigate the effect of irradiated human lung fibroblasts (HLFs) on the canonical Wnt/β-catenin signaling pathway in human umbilical cord mesenchymal stem cells (HUMSCs).

METHODSHUMSCs were cultured alone (single group) or co-cultured with HLFs exposed to 5Gy X-rays (co-culture group). The protein levels of GSK-3β, p-GSK-3β, FRAT1 and β-catenin in HUMSCs were examined by Western blotting 3 days after culture or co-culture. WISP-1 protein levels in conditioned medium were examined by ELISA.

RESULTSThe levels of p-GSK3β/GSK3β (0.15 ± 0.05), FRAT1 (0.48 ± 0.07) and β-catenin (0.50 ± 0.07) in co-cultured HUMSCs significantly decreased compared to those in single group (0.55 ± 0.05, 1.16 ± 0.13 and 2.39 ± 0.15, all P<0.05). The supernatant level of WISP-1 in co-culture group was significantly decreased [(602.23 ± 161.47) ng/mL], compared to the single group [(977.77 ± 110.56) ng/mL, P<0.05].

CONCLUSIONIrradiated HLFs attenuate the activation of canonical Wnt/β-catenin signaling pathway in HUMSCs in vitro.

CCN Intercellular Signaling Proteins ; metabolism ; Cells, Cultured ; Coculture Techniques ; Fibroblasts ; cytology ; radiation effects ; Gamma Rays ; Glycogen Synthase Kinase 3 ; metabolism ; Glycogen Synthase Kinase 3 beta ; Humans ; Intracellular Signaling Peptides and Proteins ; metabolism ; Mesenchymal Stromal Cells ; metabolism ; Proto-Oncogene Proteins ; metabolism ; Umbilical Cord ; cytology ; Wnt Signaling Pathway ; X-Rays ; beta Catenin ; metabolism

BACKGROUNDArrhythmogenic right ventricular cardiomyopathy (ARVC) is a heritable cardiac disease predominantly caused by mutations in desmosomal protein genes. Previous genetic analyses of the Chinese ARVC population are limited to small size and restriction to a single gene. This study was aimed to investigate the genotype in a large series of Chinese patients with ARVC through comprehensively screening nine ARVC-causing genes.

METHODSA total of 100 unrelated ARVC patients and 300 age, gender and ethnicity matched healthy controls were genetically tested with multiplexing targeted resequencing for nine previously reported ARVC-causing genes, including plakophilin-2, desmoplakin, desmoglein-2, desmocollin-2, plakoglobin, transforming growth factor beta-3, transmembrane protein 43, desmin and Lamin A/C.

RESULTSFifty-nine mutations were identified in 64% of the patients, among which, 93% were located in desmosomal protein genes. Plakophilin-2 mutations accounted for 54% of the total and 58% of the desmosomal mutations, with a truncating mutation type making up about 2/3 of the plakophilin-2 mutations. Only four mutations were found in non-desmosomal genes; two in transmembrane protein 43 and two in transforming growth factor beta-3. Two of them (one of each gene) appeared as single missense mutations. No mutation was identified in desmin or Lamin A/C. Multiple mutations were found in 23% of the patients, with plakophilin-2 being found in 57% of the multi-mutation carriers.

CONCLUSIONSPlakophilin-2 was the most common gene mutation that was identified in Chinese ARVC patients. Non-desmosomal genes should be added to desmosomal protein genes when performing molecular genetic screening in patients with suspected ARVC.

Adult ; Arrhythmogenic Right Ventricular Dysplasia ; genetics ; metabolism ; Asian Continental Ancestry Group ; Desmin ; genetics ; Desmoglein 2 ; genetics ; Female ; Humans ; Male ; Middle Aged ; Mutation ; Plakophilins ; genetics ; Young Adult ; gamma Catenin ; genetics

During a search for keratinocyte differentiation-related genes, we obtained a cDNA fragment from the 5'-untranslated region of a previously identified splicing variant of desmoglein 3 (Dg3). This transcript encodes a protein of 282 amino acids, which corresponds to the N-terminal truncated intracellular domain of Dg3 (Delta NDg3). Northern blot analysis detected a 4.6-kb transcript matching the predicted size of Delta NDg3 mRNA, and Western blot analysis with an antibody raised against the Dg3 C-terminus (H-145) detected a 31-kDa protein. Increased Delta NDg3 expression was observed in differentiating keratinocytes by RT-PCR and Western blot analysis, suggesting that Delta NDg3 is indeed a differentiation-related gene product. In immunohistochemical studies of normal and pathologic tissues, H-145 antibody detected the protein in the cytoplasm of suprabasal layer cells, whereas an antibody directed against the N-terminal region of Dg3 (AF1720) reacted with a membrane protein in the basal layer. In addition, Delta NDg3 transcript and protein were upregulated in psoriatic epidermis, and protein expression appeared to increase in epidermal tumors including Bowen's disease and squamous cell carcinoma. Moreover, overexpression of Delta NDg3 led to increased migration and weakening of cell adhesion. These results suggest that Delta NDg3 have a role in keratinocyte differentiation, and that may be related with tumorigenesis of epithelial origin.
Cell Adhesion ; *Cell Differentiation ; Cell Movement ; Cells, Cultured ; Desmoglein 3/*genetics/*metabolism ; Epidermis/cytology ; Gene Expression ; Humans ; Keratinocytes/*cytology ; Skin Diseases/genetics/metabolism ; gamma Catenin/metabolism

OBJECTIVE: To investigate the differential expression of junctional proteins in the normal and preeclamptic human placenta and the effect of ginsenoside Rk1 in junctional proteins. METHODS: Placental tissues from 10 women with severe preeclampsia and 5 normal women were collected at the time of their cesarean section. Five of 10 preeclamptic women were complicated with intrauterine growth restriction (IUGR). Immunohistochemistry and Western blotting was employed to localize junctional proteins (zo-1, occludin and plakoglobin) positive cells. The placental explant culture was performed to investigate if Rk1 can attenuate the expression of junctional proteins (zo-1, occluding and plakoglobin) induced by deferoxamine-induced hypoxia. Rk1 was treated at the day 3 and Western blot analysis was performed for protein quantification. RESULTS: There was no different expression of zo-1 and plakoglobin among all the study groups. Occludin showed negative at the endothelial cells of the terminal villi in both normal and preeclampsia groups. At the endothelial cells of the stem villi, occludin was detected in both normal and severe preeclamptic placenta with normal fetal growth. However, severe preeclampsia with IUGR were decreased expression of occludin at the endothelial cells of the stem villi. When we administered Rk1 to the placenta treated with DFO, expression of occludin was not different. CONCLUSION: The placental expression of zo-1 and plakoglobin were not different among the study groups, while that of occludin was significantly decreased at the endothelium of stem villi in severe preeclampsia with IUGR. Rk-1 showed no effect on the placental junctional proteins. These results suggest that occludin may play a role in pathophysiology of fetal growth restriction in utero.
Anoxia ; Blotting, Western ; Cesarean Section ; Endothelial Cells ; Endothelium ; Female ; Fetal Development ; Fetal Growth Retardation ; gamma Catenin ; Ginsenosides ; Humans ; Immunohistochemistry ; Occludin ; Placenta ; Pre-Eclampsia ; Pregnancy ; Proteins

BACKGROUND: Catenins are the associated protein with E-cadherin in the formation of adhesion complexes in normal and tumor cells related with epithelial differentiation and development of organ formation as well as in the tumor spread. The present study was aimed to find the distribution of alpha- and gamma-catenins in fetal skin development. OBJECTIVE: The purpose of this study was to observe the distribution of above two adhesion related proteins in the fetal skin during development, and to find its relationship by expression and their distribution pattern. METHODS: Skin was obtained from the scalp, chest, and sole of 21 human fetuses, ranging from 13 to 37 weeks of gestational age. Immunohistochemical staining was performed by the avidin biotin peroxidase complex method on paraffin embedded tissue using the anti-human monoclonal antibody against the human alpha- and gamma-catenins. RESULTS: alpha- and gamma-catenins were expressed strongly in basal cells of the epidermis and germ cells of skin adnexa, such as hair and eccrine glands at 13th week, followed by decreased basal cell expression. Increase in the suprabasal epithelium and differentiated adnexal epithelium, such as outer root sheath cells and eccrine ducts and glands at 18th week, and adult pattern in 23th week of gestation. Both showed similar distribution pattern in skin though gamma-catenin appeared two or three weeks later. alpha- and gamma-catenins are expressed not only in the epithelium of the skin, but also in the mesenchymal cells such as endothelial cells and fibroblasts. Though both catenins are more strongly expressed in the membrane portion, cytoplasmic expression is also noted. CONCLUSION: Both alpha- and gamma-catenin showed basically the same expression distribution pattern in the fetal skin developmental stage, suggesting that both adhesion molecules are highly related to each other in function and development of epidermis and adnexae of the skin in fetal stage.
Adult ; Avidin ; Biotin ; Cadherins ; Catenins ; Cytoplasm ; Eccrine Glands ; Endothelial Cells ; Epidermis ; Epithelium ; Fetus ; Fibroblasts ; gamma Catenin* ; Germ Cells ; Gestational Age ; Hair ; Humans ; Membranes ; Paraffin ; Peroxidase ; Pregnancy ; Scalp ; Skin* ; Thorax

BACKGROUND AND OBJECTIVES: E-cadherin and catenins (alpha, beta, gamma, p120cat) are important epithelial adhesion molecules in normal epithelial cells. Loss of E-cadherin-catenin adhesion is an important step in the progression of epithelial cancers such as tongue cancer. E-cadherin and catenins expression in carcinoma of human tongue was evaluated in relation to their clinicopathological features and prognostic values. SUBJECTS AND METHOD: Thirty-nine specimens of tongue squamous cell carcinoma were examined in this study. These patients were all treated by primary surgery without prior radiotherapy or chemotherapy. The specimens of formalin-fixed and paraffin-embedded tumor tissues were investigated by immunohistochemical analysis using E-cadherin and catenin (alpha, beta, gamma, p120cat) monoclonal antibodies. RESULTS: The expressions of E-cadherin, alpha-catenin, beta-catenin, gamma-catenin and p120cat in cell membranes were reduced or absent in 71.8%, 74.4%, 76.9%, 59.0% and 82.1% of the tumors examined, respectively. The reduced expressions of alpha-catenin and gamma-catenin in the cell membranes was cor-related with tumore differentiation (p=0.018, p=0.004, respectively). There were significant correlations between E-cadherin and expressions of the four cantenins in the cell membranes of tongue cancer. There were no correlations between beta-catenin and p120cat expression in the cytoplasm, cell nucleus and clinicopathological features. There was significant correlation between E-cadherin expression and Kaplan-Meier survival curves. CONCLUSION: These results suggest that E-cadherin and catenins (alpha, beta, gamma, p120cat) can be used as prognostic markers of human tongue squamous cell carninoma. The result of beta-catenin and p120cat absence in the nucleus suggests that Wnt/Wingless signaling or Kaiso transcription did not occur in the human tongue squamous cell carcinoma.
alpha Catenin ; Antibodies, Monoclonal ; beta Catenin ; Cadherins* ; Carcinoma, Squamous Cell ; Catenins ; Cell Membrane ; Cell Nucleus ; Cytoplasm ; Drug Therapy ; Epithelial Cells ; gamma Catenin ; Humans ; Kaplan-Meier Estimate ; Prognosis ; Radiotherapy ; Tongue Neoplasms* ; Tongue*

PURPOSE: The significance of abnormal E-cadherin/ catenin complex expression and the correlation of each of its components in cancer remain unclear. This study aimed to characterize the clinical significance of the abnormal membrane expression of the E-cadherin/ catenin complex and the localization patterns of the beta- catenin and p120CTN in early gastric cancer. MATERIALS AND METHODS: Immunohistochemical staining for E-cadherin, alpha-, beta- and gamma-catenin and p120CTN were performed on 47 early gastric cancer specimens. The patterns of membrange expression of the E-cadherin/catenin complex, and the localization patterns of the beta-catenin and p120CTN, were semi quantitatively graded as loss, reduced, preserved or negative and positive. RESULTS: An abnormal immunoreactivity of at least one of E-cadherin/catenin complex proteins was noted in 46 (97.8%) of the 47 early gastric cancer cases. There were no significant correlations of the membrane E-cadherin/catenin expression with, either, sex, age, location, size, macroscopic type, depth of invasion or lymphovascular invasion. Abnormal expressions of membrane E-cadherin, beta-catenin and gamma-catenin were more frequent in the diffuse-type than in the intestinal type. No linear correlation was shown for the beta-catenin between the membrane and cytoplasmic expressions. Nuclear staining of the beta-catenin was observed in 5 (10.6%) cases, but nuclear staining of the p120CTN, a promotor of Kaiso transcriptional factor, was not seen. CONCLUSION: These results suggest that alterations of the E-cadherin/catenin complex may be involved in the early stages of gastric cancer. Although beta-catenin functions as a transcriptional factor, the inactivation of membrane E-cadherin does not appear to result in significant increases in the level of cytoplasmic beta-catenin. Kaiso transcriptional factor may not be involved in the early carcinogenesis of gastric cancer.
beta Catenin ; Cadherins ; Carcinogenesis ; Cell Adhesion ; Cytoplasm ; gamma Catenin* ; Membranes ; Stomach Neoplasms*