ABSTRACT
Beta-catenin/TCF signaling has been reported to promote the growth and metastasis of pancreatic cancer cells. However, the regulation for the beta-catenin/TCF transcriptional complex remains largely unknown. Here, we have found that YEATS4 is a positive regulator for Beta-catenin/TCF signaling. The expression of YEATS4 was elevated in clinical pancreatic cancer samples and pancreatic cancer mouse model. Up-regulation of YEATS4 promoted the growth, migration and invasion of pancreatic cancer cells, while knocking down the expression of YEATS4 inhibited the growth, migration, invasion and metastasis of pancreatic cancer cells. Moreover, the mechanism study revealed that YEATS4 interacted with beta-catenin and activated beta-catenin/TCF signaling. Furthermore, knocking down the expression of YEATS4 impaired the malignant transformation of normal pancreatic cells (HPDE6C7) by the oncogenic Ras. Taken together, our study demonstrated the oncogenic roles of YEATS4 in the progression of pancreatic cancer by activating beta-catenin/TCF signaling and suggested that YEATS4 might be a promising therapeutic target for pancreatic cancer.
Subject(s)
Cell Transformation, Neoplastic/genetics , Cell Transformation, Neoplastic/metabolism , Pancreatic Neoplasms/genetics , Pancreatic Neoplasms/metabolism , Signal Transduction , TCF Transcription Factors/metabolism , Transcription Factors/genetics , beta Catenin/metabolism , Animals , Cell Line, Tumor , Cell Movement , Cell Proliferation , Disease Models, Animal , Gene Expression Regulation, Neoplastic , Heterografts , Humans , Mice , Mice, Transgenic , Pancreatic Neoplasms/pathology , Tumor Stem Cell AssayABSTRACT
<p><b>OBJECTIVE</b>The aim of this study was to explore the effect of SPARC on the anti-cancer effect of gemcitabine and underlying mechanism in pancreatic cancer.</p><p><b>METHODS</b>After treating with gemcitabine, the proliferation rate of MIA PaCa2, MIA PaCa2/V and MIA PaCa2/SPARC69 cells was detected by MTT assay. The cell cycle distribution and cell apoptosis in each group were examined by flow cytometry, and the capability of clone formation was tested by adhesion-dependent clone formation assay. The apoptosis-related proteins were analyzed by Western blot.</p><p><b>RESULTS</b>The growth of pancreatic cancer cells was inhibited by gemcitabine in a time-dependent and dose-dependent manner. Its IC50 at 24, 48, and 72-h was (40.1 ± 2.5) µmol/L, (15.0 ± 0.5) µmol/L and (6.6 ± 0.1) µmol/L, respectively. The overexpression of SPARC increased the inhibitory effect of gemcitabine on growth of pancreatic cancer MIA PaCa2/SPARC69 cells, presenting a dose- and time- dependent manner. Its IC50 at 24, 48, 72 h was (24.3 ± 1.5) µmol/L, (7.7 ± 0.3) µmol/L and (4.8 ± 0.2) µmol/L, respectively. The clone formation assay showed that before gemcitabine treatment, the clone numbers of MIA PaCa2, MIA PaCa2/V and MIA PaCa2/SPARC69 cells were (2350 ± 125), (2130 ± 120) and (1567 ± 11), respectively. After gemcitabine treatment, the clone numbers of MIA PaCa2, MIA PaCa2/V and MIA PaCa2/SPARC69 cells were ( 1674 ± 79) , (1587 ± 94) and (557 ± 61), respectively. The overexpression of SPARC enhanced the chemosensitivity of MIA PaCa2 cells to gemcitabine chemotherapy. After treating with 10 µmol/L gemcitabine for 48 h, the ratio of G0/G1 cells in MIA PaCa2, MIA PaCa2/V and MIA PaCa2/SPARC69 cells were (56.0 ± 5.5)%, (55.0 ± 4.5)% and (68.0 ± 7.0)%, respectively. The cells arrested at G0/G1 phase were significantly increased in the MIA PaCa2/SPARC69 cells. The apoptosis rates of MIA PaCa2, MIA PaCa2/V and MIA PaCa2/SPARC69 cells were (22.4 ± 2.5)%, (19.9 ± 2.0)% and (37.7 ± 3.9)%, respectively, indicating that overexpression of SPARC enhanced the gemcitabine-induced apoptosis in MIA PaCa2 cells. The Western blot analysis showed that, compared with MIA PaCa2 and MIA PaCa2/V cells, the expression of caspase-2, -8, -9 and cleaved PARP protein was significantly increased, while the expression of Bcl-2 was not changed significantly in the MIA PaCa2/SPARC69 cells.</p><p><b>CONCLUSION</b>SPARC can enhance the chemosensitivity of pancreatic cancer cells to gemcitabine via regulating the expression of apoptosis-related proteins.</p>
Subject(s)
Humans , Antimetabolites, Antineoplastic , Pharmacology , Apoptosis , Caspase 2 , Metabolism , Caspase 8 , Metabolism , Caspase 9 , Metabolism , Cell Cycle , Cell Cycle Checkpoints , Cell Line, Tumor , Cell Proliferation , Cysteine Endopeptidases , Metabolism , Deoxycytidine , Pharmacology , Dose-Response Relationship, Drug , Drug Resistance, Neoplasm , Osteonectin , Metabolism , Pancreatic Neoplasms , Metabolism , Pathology , Poly(ADP-ribose) Polymerases , Metabolism , Time FactorsABSTRACT
EphB4 tyrosine kinase receptor has been involved in various physiologic and pathologic processes, and the role of the EphB4 in tumorigenesis has recently attracted much interest. However, its function in papillary thyroid carcinoma remains poorly understood. In this study, we explored the function of EphB4 in papillary thyroid carcinoma. We found that the expression of EphB4 was significantly upregulated in clinical samples. Overexpression of EphB4 in papillary thyroid carcinoma cell lines accelerated cell migration. In contrast, downregulation of EphB4 inhibited cell migration and suppressed in vivo tumor metastasis. Furthermore, we showed that EphB4 promoted cell migration by inhibiting the phosphorylation of FAK and paxillin. Moreover, EphB4 promoted cell migration in a kinase-independent manner. Taken together, our findings suggest that EphB4 plays an important role in the progression of papillary thyroid carcinoma by stimulating cell migration and EphB4 might be a potential therapeutic target in papillary thyroid carcinoma.
Subject(s)
Carcinoma/genetics , Cell Movement/genetics , Gene Expression , Receptor, EphB4/genetics , Thyroid Neoplasms/genetics , Animals , Carcinoma/metabolism , Carcinoma/pathology , Carcinoma, Papillary , Cell Line, Tumor , Gene Silencing , Humans , Mice , Neoplasm Metastasis/genetics , Phosphotransferases/metabolism , Receptor, EphB4/metabolism , Thyroid Cancer, Papillary , Thyroid Neoplasms/metabolism , Thyroid Neoplasms/pathology , Xenograft Model Antitumor AssaysABSTRACT
Objective To study the effects of elodronate-liposome on inducing apoptosis of alve-olar macrophages from rats with acute neetotizing pancreatitis (ANP).Methods The AMs of eight rats with ANP were isolated, purified then incubated from broehoalveolar lavage by the differing rates of attachment of the various cell types in a forty-well cell culture plate.Then they were randomized in-to five groups including control group,blank liposome group( 50 μ1, 100 μ1),clodronate-liposome group (50μ1,100μ1).Values of OD were determined by MTT.AO fluorescence and haematoxylin dye were employed to determine the apoptosis of the AMs.Results There were no significant differences be-tween control group and blank liposome group(50 μ1, 100 μ1).Significant differences were found be-tween control group and clodronate-liposome group(50 μ1, 100 μ1).There were no marked differences between blank liposome group(50μ1, 100 μ1)and clodronate-liposome group(50 μ1,100μ1).AO fluo-rescence and haematoxylin dye were available to define the apoptosis of the AMs.Conclusion Clodr-onate-liposome can effectively induce the apoptosis of the AMs.
ABSTRACT
OBJECTIVE:To study the dynamic changes of TNF-?/IL-10in model rats with acute necrotizing pancreatitis (ANP)and to study the intervention outcome of salvia miltiorrhiza.METHODS:A total of96rats were randomly divided into three groups(each with32rats):pancreatitis(P)group,salvia miltiorrhiza treatment(T)group and control group(C),the blood samples of rats in each group were taken to determine levels of TNF-?and IL-10and the ratio changes of TNF-?/IL-10.Meanwhile,pancreas tissue sample was collected for pathological scoring.RESULTS:Group P had significantly higher serum levels of TNF-?and IL-10than did group C(P
ABSTRACT
Foods derived from plants are thought to be devoid of vitamin B12. We have found that some fermented foods contained microbiologically active vitamin B12 components.In order to ascertain the amount of physiological active vitamin B12, we have established a bioautographic area quantitative method of determination of vitamin B12. The procedure is described briefly as follows:The extract of fermented soybean food sample is spotted on a filter paper strip. The Paper is developed with a mixture of secbutanol-water-ammonia water- 5 % potassium cyanide (100: 50: 1: 0.5) using descending technique. After developing for 24-72 hours, the strip is taken out and dried in air. It is then put on the surface of nutrient agar containing E. coli 44110 and 2,3,5,-triphenyl tetrazolium chloride. After incubation, the area of spot of growth is measured and the amount of vitamin B12 in the sample is calculated.Two types of fermented soybean food samples were assayed by the above procedure, mean values of vitamin B12 content are: fermented soybean curd with strong smell contains 0.97 ?g/100g sample, fermented soybean curd contains 0.41 ?g/100g sample. Compared with animal foods, both types of fermented soybean foods may be considered as good source of vitamin B12.
