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Objective:To observe the expression patterns of miR-195 and DLL4 in colorectal cancer, and to explore the relationship between miR-195 and DLL4 and clinicopathological parameters and prognosis of patients with colorectal cancer.Methods:The relative expression of miR-195 in 56 colorectal cancer tissues was detected by real-time fluorescent quantitative PCR the expression of DLL4 protein and tumor microvessel density (MVD) were detected by bloting and immunohistochemistry. Colon cancer cell line SW480 was treated with miR-195. The expression of DLL4, Jagged1, (the intracellular domain of notch, NICD), CyclinD1, Hes1, Bcl-2 and NF-kB were detected by bloting.Results:The expression level of DLL4 protein in colorectal cancer tissues was significantly higher than that in normal intestinal mucosa (30/56 vs.16/56, t=5.323, P=0.018). The expression level of miR-195 was significantly lower than that in normal intestinal mucosa (36/56 vs.20/56, t=2.371, P=0.008). The expression of DLL4 was negatively correlated with the expression of miR-195 ( r=- 0.881, P=0.015) , which was closely related to the differentiation degree, lymph node metastasis and TNM stage . The prognosis of patients with high expression of DLL4 was significantly worse than that with low expression of DLL4 ( P=0.013). The prognosis of patients with low expression of miR-195 was worse than that with high expression of miR-195 ( P=0.009) . Conclusion:The antagonistic expression of miR-195 and Notch might be closely related to the occurrence and development of colorectal cancer, which can be used as a new reference index for prognosis of colorectal cancer.
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Objective To investigate the relationship between the expression of suppressor of zeste 12(SUZ12) and the clinicopathological parameters and prognosis in patients with gastric cancer. Methods SUZ12 protein expression levels in 97 cases of resected gastric cancer were detected by immunohitochemistry method, the relations between SUZ12 expression levels and the survival were estimated by Kaplan-Meier curve. Results The positive rate of SUZ12 expression in gastric cancer tissues was 43%, significantly higher than that (15%)in the adjacent noncancerous tissues( P = 0. 002). SUZ12-positive expression was significantly correlated with tumor differentiation ( P = 0. 018 ), lymph nodes metastasis ( P = 0. 023 ) and TNM staging(P = 0. 014). Gastric cancer patients with SUZ1 2-positive expression had worse prognosis than those with SUZ12-negative expression ( P = 0. 024). Conclusions SUZ12 is overexpressed in tissues of gastric carcinoma, SUZ12 is an independent prognosis factor of patients with gastric carcinoma.
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Objective To investigate the expressions of Dapper1 in gastric carcinoma and elucidate its relationship with survivin and its role in tumor cell apoptosis. Methods Dapper1 mRNA was detected with RT-PCR using specimens from 30 cases of gastric carcinoma and the corresponding normal gastric mucosa.The pcDNA3.1-Dpr1 plasmid was transfected into SGC-7901 cells with LipofectamineTM 2000.The effect of upregulation of Dpr1 on SGC7901 cell apoptosis was determined by flow cytometry.The downregulation of survivin、Dvl-2 and β-catenin protein expression were detected by Western blot analysis.Results Downregulation of Dpr1 gene expression was observed in 17(57%)of 30 human gastric cancer and the downregulation was significantly correlated with the depth of invasion and the degree of differentiation (P<0.05).Also,upregulation of Dpr1 mRNA and downregulation of survivin mRNA were detected after transfecting pcDNA3.1-Dpr1 plasmid in SGC7901 cells,which led to downregulation of survivin、Dvl-2、β-catenin protein and increase of the SGC7901 cell apoptosis rate from 2.89%to 13.96%.Conclusion Downregulation of Dp1l gene expression is common in human gastric carcinoma,and upregulation of Dpr1 results in significant inhibition of survivin expression which can induce apoptosis of SGC7901 cells.
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<p><b>OBJECTIVE</b>To investigate the expressions of cyclin E, cyclin dependent kinase 2 (CDK-2) and cyclin-dependent kinase inhibitor p57(KIP2) in human gastric cancer, and to evaluate the relationships between protein levels and clinicopathological parameters.</p><p><b>METHODS</b>Western blot was used to measure the expressions of cyclin E, CDK-2 and p57(KIP2) proteins in the surgically resected gastric carcinoma, adjacent normal mucosa and metastatic lymph nodes from 36 patients.</p><p><b>RESULTS</b>Cyclin E and CDK-2 protein levels were higher in gastric cancer tissues in comparison with normal tissues (P < 0.05). Overexpression of cyclin E was correlated with lymph node involvement, poor histological grade and serosa invasion (P < 0.05). Overexpression of CDK-2 was correlated with lymph nodes involvement (P < 0.05). No statistically significant difference between cyclin E and CDK-2 expression was found when samples were stratified according to tumor size (P > 0.05). Expression of cyclin E and CDK-2 showed a positive linear correlation (r = 0.451, P = 0.01). Protein levels of p57(KIP2) were lower in gastric cancer tissues than in the normal mucosa (P < 0.05). Decreased expression of p57(KIP2) was correlated with lymph node involvement (P < 0.05). No statistically significant difference in p57(KIP2) expression was found when sample were stratified according to tumor size, histological grade or serosa invasion (P > 0.05). In metastatic lymph nodes, expression of cyclin E was increased and the expression of p57(KIP2) decreased.</p><p><b>CONCLUSION</b>Overexpressions of cyclin E, CDK-2 and downregulated expression of p57(KIP2) may play important roles in tumorigenesis and metastatic potential of gastric cancer.</p>
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Humanos , Western Blotting , Quinases relacionadas a CDC2 e CDC28 , Ciclina E , Fisiologia , Quinase 2 Dependente de Ciclina , Inibidor de Quinase Dependente de Ciclina p57 , Quinases Ciclina-Dependentes , Fisiologia , Metástase Linfática , Proteínas Nucleares , Fisiologia , Proteínas Serina-Treonina Quinases , Fisiologia , Neoplasias Gástricas , Química , PatologiaRESUMO
<p><b>OBJECTIVE</b>To investigate the MEK and ERK expression and their relationship with clinicopathological parameters in human breast carcinoma, and the effect of preoperative chemotherapy on MEK and ERK protein expression.</p><p><b>METHODS</b>Samples were obtained from 56 patients with breast carcinoma and 8 patients with benign tumors. Sixteen of the 56 patients received preoperative chemotherapy. Western blot and immunohistochemistry were used to measure the expression of MEK1, MEK2 and ERK1, ERK2 protein.</p><p><b>RESULTS</b>MEK2 and ERK1, ERK2 protein levels were increased in breast carcinoma tissue compared with those in adjacent normal tissues (t = 7.244, 5.959, 3.735, P < 0.01) and benign tumors (t = 2.206, P < 0.05). The levels of MEK1 were decreased. The expression of MEK2 protein in ER negative patients was higher than that in ER positive ones. MEK2 protein levels were lower in patients who received preoperative chemotherapy than in those who did not.</p><p><b>CONCLUSION</b>Overexpression of MEK-ERK may play an important role in the development of human breast carcinoma. MEK and ERK protein expressions are inhibited by preoperative chemotherapy.</p>
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Adulto , Idoso , Feminino , Humanos , Pessoa de Meia-Idade , Western Blotting , Neoplasias da Mama , Diagnóstico , Metabolismo , Imuno-Histoquímica , MAP Quinase Quinase 1 , MAP Quinase Quinase 2 , Sistema de Sinalização das MAP Quinases , Fisiologia , Proteína Quinase 1 Ativada por Mitógeno , Metabolismo , Proteína Quinase 3 Ativada por Mitógeno , Quinases de Proteína Quinase Ativadas por Mitógeno , Metabolismo , Proteínas Quinases Ativadas por Mitógeno , Metabolismo , Prognóstico , Proteínas Quinases , Metabolismo , Proteínas Serina-Treonina Quinases , Metabolismo , Proteínas Tirosina Quinases , MetabolismoRESUMO
Objective This study was to investigate the role of STAT3 signaling pathway in gastric cancer invasiveness. Methods Janus kinase 2 selective inhibitor AG490 was used to inhibit the activation of STAT3 signaling pathway. Matrigel-coated transwell was employed to estimate the invasiveness of BGG-823 cells. Alterations in the activity of STAT3 and expression of MMPs and TIMPs were measured by Western blot and RT-PCR. Results The activation of STAT3 signaling pathway was suppressed by AG490. AC490 significantly decreased the invasiveness of gastric cancer cell line BGC-823 through Matrigel-coated filters. Decreased invasion was associated with the inhibition of MMP-2, and MMP-9 at both mRNA and protein levels by RT-PCR and Western blot. Meanwhile, the expression of TIMP-1, and TIMP-2 were up-regulated at mRNA levels. Conclusion Inhibition of STAT3 signaling pathway in AG490 suppresses gastric cancer cell invasiveness through down-regulation of several invasion-related factors including MMP-2, and MMP-9 and up-regulation of their inhibitors.
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Objective This study was to investigate the relationship between the activities of thymidine phosphorylase (TP) and dihydropyrimidine dehydrogenase (DPD) and clinicopathological parameters in human colorectal cancer,and the relationship between TP and/or DPD activity in tumor tissue and efficiency of chemotherapy. MethodsSixty-eight patients undergoing surgery for primary colorectal cancer were enrolled including 40 patients receiving postoperative adjuvant chemotherapy with 5-FU plus leucovorin(de Gramont regimen). The activities of TP and DPD both in tumor tissue and in normal tissue were determined by enzyme-linked immunosorbent assay(ELISA). Results The TP activity was significantly higher in tumor than in normal tissues(P
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Objective The purpose of this study was to analyze the alterations of T_1、T_2 lymphocyte subsets in the peripheral blood of patients with colorectal cancer. MethodsTwenty patients with primary colorectal cancer were enrolled into this study, T_1 and T_2 in the peripheral blood were evaluated by detecting the intracellular interferon-? and interleukin-4 production with 4-color flow cytometry. ResultsThe percentage of T_1 and T_2 in the peripheral blood of cancer patients was lower significantly than healthy controls [(36?11)% and 3.3(1.9)% vs. (46?12)% and 4.1(3.1)%](P
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Objective To detect the antitumor activity against SW620 and syngenic colon cancer SW480 by a fusion of human metastatic colon cancer SW620 cells and human peripheral blood-derived dendritic cells (DCs). Methods SW620 cells and human peripheral blood- derived DCs were fused with polyethylene glycol(PEG). The fusion cells were confirmed by "phenotypes determine, morphologic observation, and cytotoxic T lymphocyte response induced by the fusion were studied. Results Mature DCs with highly expressed surface markers (HLA-ABC, HLA-DR and CD80, CD86, CD83) were generated in vitro. The fusion of SW620 cells with DC resulted in a hybrid cell with morphologic as well as phenotypic characteristics of both DC and tumor cells. Flow cytometry showed that the highest fusion efficiency was 27. 12%. CTL assay demonstrated that the DC/SW620 fusion induced specific cytotoxic responses against the SW620 and SW480 cells. Conclusion The fusion of tumor cells with DCs induces tumor rejection.
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Objective To study the influence of transferring a dominant-negative Stat3 gene, Stat3?on colon cancer cells' proliferation and apoptosis in vitro. Methods Cell culture of human colon cancer cell line SW480 and transient transfection were used to evaluate the effect of transferring Stat3?to cancer cells. Cell proliferation, cell cycle and apoptosis were quantified by MTT and flow cytometry, respectively. The mRNA expression of Stat3's target gene cyclin D1 and bcl-xL was detected by reverse transcription polymerase chain reaction. Independent t tests were used for data statistics. Results 36 h after Stat3?plasmids transfection, proliferation of SW480 cells was significantly inhibited (t =5. 216,P = 0.006); cell proportion of G0/G1 phase increased from 40.37% to 67.25% and early apoptosis cells increased from 5. 34% to 24. 42% ; mRNA expression of cyclin Dl and bcl-xL declined significantly (t = 5.288,P=0.010;t=3.517,P=0.025). Conclusion Blocking Stat3 signaling pathway by transfection of Stat3?plasmid inhibits the proliferation and promotes apoptosis of colon cancer cells, which provides a experimental foundation of Stat3 targeted colon cancer gene therapy.
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Objective The aim of this study is to investigate STAT3 expression and the relationship between protein levels of STAT3 and clinicopathological parameters together with preoperative chemotherapy in human breast cancer. Methods Samples were obtained from 51 patients with breast carcinoma in which 7 patients had received preoperative chemotherapy. Western blot was used to measure the protein expression in tissue and SK-BR-3 cell line. MTT method was used to evaluate the proliferation. Cell-cycle and apoptosis were quantified by flow cytometry. Results STAT3 protein levels increased in breast cancer tissues compared with adjacent normal tissues(t=5.540,P
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Objective To evaluate the clinical significance of combined serum tumor markers assay in patients with gastric or colorectal carcinoma. Methods Serum level of 12 common tumor markers, including CA19-9,NSE,CEA,CA242 Jerritin,Beta-HCG,AFP,free-PSA,PSA,CA125,HGH and CA153, was measured with multi-tumor markers protein biochip detective system in 179 cases of gastric and colorectal carcinoma, 82 patients with benign digestive disease and 160 healthy volunteers. Results Cancer patients had significantly higher positive rates than that of two other controls (P
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Objective To determine the correlation between intestinal epithelial cell apoptosis; and STAT3 activation during murine severe acute pancreatitis (SAP).Methods Wistar rats were divided into four groups (12 in each group): S (sham), C (control), A (SAP), N (NAC was given 1 hour before SAP).Samples were obtained 3 or 24 hours after for the determination of plasma D-lactate, intestinal mucosa MDA and XO, immunohistochemical staining for PCNA.Apoptosis of the epithelial cell was examined with TUNEL and DNA laddering.The expressions of Bcl-2? Bax? p-STAT3 in intestinal mucosa were assessed by Western blot.Results Plasma D-lactate and intestinal epithelial cell apoptosis increased in group A, especially at 24 hours after operation ( P
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Objective To investigate the role of p38? pathway and HO-1 in the genesis and chemotherapy resistance of breast cancer. Methods The proliferation and apoptosis of human breast cancer cells were examined by MTT assay. The expression of p38? and HO-1 mRNA were examined by RT-PCR. Results The p38? mRNA level in 78% of samples was significantly greater than that in the normal tissue and the p38? mRNA level in patients with lymph node metastasis was higher than that without lymph node metastasis (P
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AIM: The purpose of the study was to examine colon cancer cell lines to determine whether Stat5b/Survivin plays an important role in the process of apoptosis in colon cancer cells. METHODS: Protein lysates were extracted from colon cancer cells. Human colon cancer cell line HT29 was transfected with Stat5b antisense oligonucleotide mediated by liposome. MTT assay was used to measure the proliferation. Flow cytometry was applied to analyze the cell cycle and apoptosis. EMSA was used to detect the activity of Stat5. Western blotting was applied to measure the expression of Stat5, p-Stat5, cyclin D1, Survivin, Bcl-2 and Bcl-xL. RESULTS: Targeting of Stat5 using antisense oligonucleotide against the translation site resulted in apoptosis and downregulaed the expressions of Stat5, p-Stat5, cyclin D1 and Survivin, but not Bcl-2 and Bcl-xL. CONCLUSION: Constitutive activation of Stat5 is associated with the carcinogenesis of colon cancer cells. Blocking of Stat5 signaling inhibits the expression of Survivin and induces apoptosis in colon cancer cells.