Objective To investigate the expressions of PTEN and Caspase-3 proteins in human breast carcinoma,and to evaluate their clinicopathological implications during the tumorigenesis and progression of breast cancer.Method...Objective To investigate the expressions of PTEN and Caspase-3 proteins in human breast carcinoma,and to evaluate their clinicopathological implications during the tumorigenesis and progression of breast cancer.Methods The expressions of PTEN and Caspase-3 proteins in 95 cases of breast cancer and 15 cases of benign breast diseases were investigated immunohistochemically.Correlations between the expression of PTEN protein,Caspase-3 protein,and clinicopathological features of breast cancers were analyzed.Results The loss expression rate of PTEN protein in tumor tissues was significantly higher than that in benign breast diseases(33.7% vs.0,P<0.01).Analysis of the clinicopathological features showed that PTEN expression level was negatively correlated with TNM stage,histological grade,axillary lymph node status,recurrence,and metastasis(P<0.05).The positive expression level of Caspase-3 was negatively correlated with TNM stage(P<0.01),but not related with histological grade,axillary lymph node status,recurrence,or metastasis(P>0.05).In addition,the expression of PTEN protein had significantly positive correlation with the expression of Caspase-3 protein in breast cancer(P<0.01).Conclusion The combination detection of PTEN and Caspase-3 may serve as an important index to estimate the pathobiological behavior and prognosis of breast cancer.展开更多
OBJECTIVE To study the expression of phosphorylated p38 mitogen-activated protein kinase （p-p38） and uPA and the correlation of their expression with breast cancer clinicopathological characteristics, and to investi...OBJECTIVE To study the expression of phosphorylated p38 mitogen-activated protein kinase （p-p38） and uPA and the correlation of their expression with breast cancer clinicopathological characteristics, and to investigate the role of the p38MAPK-signaling pathway in regulating uPA expression in breast cancer cells.METHODS Immunohistochemistry （S-P） was used to test the expression of p-p38 and uPA in 60 specimens of breast cancer tissues. Western blots were adopted to detect expression of the p-p38 and uPA proteins in MDA-MB-231 and MCF-7 breast cancer cells, and uPA expression after treatment with SB203580, a specific inhibitor of p38 MAPK.RESULTS The positive rate of the p-p38 protein and uPA protein expression in the breast cancer tissues was 56.7% and 60.0%,respectively. The expression of p-p38 was positively related to the expression of uPA （r = 0.316, P 〈 0.05）. The expression of p-p38 and uPA was related to lymph node metastasis and the TNM stage （P 〈 0.05）, but it was not related to the patient＇s age or tumor size （P 〉 0.05）. The expression of p-p38 and uPA in MDA- MB-231 cells was higher than that in MCF-7 cells. SB203580 inhibited the p38 MAPK pathway and reduced uPA protein expression.CONCLUSION The p38 MAPK-signaling pathway promotes breast cancer malignant progression by up-regulating uPA expression ,and it may be an important process in breast cancer invasion and metastasis.展开更多
The matrix metalloproteinases (MMPs) are a family of zinc-dependent endopeptidases that play a key role in both normal and pathological processes involving tissue remodeling events. The expression of these proteolytic...The matrix metalloproteinases (MMPs) are a family of zinc-dependent endopeptidases that play a key role in both normal and pathological processes involving tissue remodeling events. The expression of these proteolytic enzymes is highly regulated by a balance between extracellular matrix (ECM) deposition and its degradation, and is controlled by growth factors, cytokines, hormones, as well as interactions with the ECM macromolecules. Furthermore, the activity of the MMPs is regulated by their natural endogenous inhibitors, which are members of the tissue inhibitor of metalloproteinases (TIMP) family. In the normal mammary gland, MMPs are expressed during ductal development, lobulo-alveolar development in pregnancy and involution after lactation. Under pathological conditions, such as tumorigenesis, the dysregulated expression of MMPs play a role in tumor initiation, progression and malignant conversion as well as facilitating invasion and motastasis of malignant cells through degradation of the ECM and basement membranes. Matrix metalloproteinases and their expression in mammary gland展开更多
OBJECTIVE To investigate the efficiency of gene silencing by CXCR4- siRNAs （small interfering RNA）, and to examine the invasive ability and the expression of other metastatic-associated genes in siRNA-treated breast...OBJECTIVE To investigate the efficiency of gene silencing by CXCR4- siRNAs （small interfering RNA）, and to examine the invasive ability and the expression of other metastatic-associated genes in siRNA-treated breast cancer cells.METHODS Three siRNAs were designed and cloned into the pSilenc ^TM 3.1-H1 neo vector. The reconstructed plasmids were purified and transfected into the T47D breast cancer cell line, which highly expressed CXCR4. The amount of CXCR4 expression in the transfected cells was measured by flow cytometry and Real-time PCR. Cell invasive ability was evaluated using 24-well Matrigel invasion chambers. In addition, the expression of other metastatic-associated genes, such as E-cad, IGFBP-5, FN and MMP-2, was assessed by Real-time PCRRESULTS The suppression rates of CXCR4 mRNA expression reached 95.7%, 85.9% and 98.3%compared with control-siRNA cells in the 3 CXCR4- siRNA T47D cells respectively. FCM assays for CXCR4 protein expression showed a similar inhibitory effect. The invasion indexes of these CXCR4- siRNA cells were 0.037, 0.290 and 0.188 respectively compared with control- siRNA cells. After treatment of the cells with CXCR4-siRNA, the expression of E-cad showed an upward tendency and that of IGFBP-5 had a downward trend, while alteration in expression of FN and MMP2 varied without a consistant effect.CONCLUSION CXCR4 plays an important role in modulating migra- tion of human breast cancer cells. Small interfering RNA can significantly silence the CXCR4 gene in the human T47D breast cancer cell line. The results of this study strengthen the need for further research on novel gene therapy against breast cancer metastasis.展开更多
OBJECTIVE To investigate the expression of the RECK gene in human breast （cancer） cell lines, and to determine the relationship between RECK gene expression and the invasive capacity of the breast cancer cell lines....OBJECTIVE To investigate the expression of the RECK gene in human breast （cancer） cell lines, and to determine the relationship between RECK gene expression and the invasive capacity of the breast cancer cell lines.METHODS The invasive capacity of breast （cancer） cell lines including HBL-100, MCF-7 and MDA-MB-435S were determined by the Tran- swell method. The protein expression levels of RECK, MMP-2 and MMP- 9 genes in these three cell lines were measured by immunocytochemical methods. The expressions of the RECK gene and protein level were measured hv RT-PCR and Westrn hints in the cell lines respectively.RESULTS The order of the nvasive capacity of the breast （cancer） cell lines was MDA-MB-435S, being the highest, and HBL-100, being the lowest. The invasive capacity difference between any two groups among the three groups was significant （P〈0.01）. The protein expression level of the RECK gene in the HBL-100 cell line was highest, and no expression was detected in MDA-MB-435S cells. Moreover, the expression of the RECK gene was negatively correlated with the expression of the MMP-2 and MMP-9 genes. The mRNA level of the RECK gene in HBL-100 cells was the highest, but no expression was found in the MDA-MB-435S cells （P〈0.001）.CONCLUSION There was a significant negative correlation between the expression level of the RECK gene and invasive capacity in vitro, and the RECK gene expression showed an inverse proportion to that of the MMP-2, MMP-9 genes.展开更多
AIM： We examined quantitative mRNA expression of growth factor receptors （c-erbB-1, c-erbB-2） and the anti-apoptosis gene survivin known to be regulated in pancreatic adenocarcinomas and compared the expression pat...AIM： We examined quantitative mRNA expression of growth factor receptors （c-erbB-1, c-erbB-2） and the anti-apoptosis gene survivin known to be regulated in pancreatic adenocarcinomas and compared the expression pattern with that in carcinomas of the papilla of Vater. METHODS： Quantitative real-time reverse transcriptase- PCR （QRT-PCR, Taqman^TM） was performed to analyze mRNA expression levels of c-erbB-1, c-erbB-2 and survivin in normal and corresponding tumor samples of 31 pancreatic adenocarcinomas and 8 cancers of the papilla of Vater. RESULTS： The overall median mRNA expression of survivin was significantly increased in both adenocarcinoma of the pancreas （P〈0.01） and papilla of Vater （P〈0.008） compared with uninvolved normal control tissue. In pancreatic cancer, expression of c-erbB-1 was significantly decreased compared with the normal pancreatic tissue （P〈0.03）, whereas in the cancer of the papilla of Vater expression of c-erbB-2 was significantly downregulated （P〈0.05） compared with the paired normal samples. Gene expression was not associated with tumor stage, differentiation or prognosis. CONCLUSION： The common anti-apoptosis gene survivin is overexpressed both in the cancer of the papilla of Vater and pancreas. In contrast, the growth factor receptor genes c-erbB-1 and c-erbB-2 are differentially regulated in both tumor entities adding further evidence that pancreatic cancer is biologically different from the cancer of papilla of Vater.展开更多
A malignant transformed mammary epithelial cell line (11A1) was transfected with liposome encapsulated eukaryotic expression plasmid pCMV-neo-RB, yielding 4 constant clones which have obvious pheno-typic reversion cha...A malignant transformed mammary epithelial cell line (11A1) was transfected with liposome encapsulated eukaryotic expression plasmid pCMV-neo-RB, yielding 4 constant clones which have obvious pheno-typic reversion changes, and named 11A1-R1-R4 respectively. Further experiments showed that the 11A1-R1 behaved like normal epithelial cells in both morphological and biological characteristics, with decreased clonogenicity in solid argar medium as well as decreased tumorigenicity. Northern blot hybridization showed increased expression of RB gene and decreased expression of c-myc gene in 11A1-R1, 11A1-R2 cells compared to 11A1 cells. This was an ideal phenotypic reversion model for epithelial transformed cell line and demonstrated that the RB gene can reexpress and suppress malignant phenotype in RB inactive cells.展开更多
基金Supported by the National Natural Science Foundation of China(30371607)
文摘Objective To investigate the expressions of PTEN and Caspase-3 proteins in human breast carcinoma,and to evaluate their clinicopathological implications during the tumorigenesis and progression of breast cancer.Methods The expressions of PTEN and Caspase-3 proteins in 95 cases of breast cancer and 15 cases of benign breast diseases were investigated immunohistochemically.Correlations between the expression of PTEN protein,Caspase-3 protein,and clinicopathological features of breast cancers were analyzed.Results The loss expression rate of PTEN protein in tumor tissues was significantly higher than that in benign breast diseases(33.7% vs.0,P<0.01).Analysis of the clinicopathological features showed that PTEN expression level was negatively correlated with TNM stage,histological grade,axillary lymph node status,recurrence,and metastasis(P<0.05).The positive expression level of Caspase-3 was negatively correlated with TNM stage(P<0.01),but not related with histological grade,axillary lymph node status,recurrence,or metastasis(P>0.05).In addition,the expression of PTEN protein had significantly positive correlation with the expression of Caspase-3 protein in breast cancer(P<0.01).Conclusion The combination detection of PTEN and Caspase-3 may serve as an important index to estimate the pathobiological behavior and prognosis of breast cancer.
文摘OBJECTIVE To study the expression of phosphorylated p38 mitogen-activated protein kinase （p-p38） and uPA and the correlation of their expression with breast cancer clinicopathological characteristics, and to investigate the role of the p38MAPK-signaling pathway in regulating uPA expression in breast cancer cells.METHODS Immunohistochemistry （S-P） was used to test the expression of p-p38 and uPA in 60 specimens of breast cancer tissues. Western blots were adopted to detect expression of the p-p38 and uPA proteins in MDA-MB-231 and MCF-7 breast cancer cells, and uPA expression after treatment with SB203580, a specific inhibitor of p38 MAPK.RESULTS The positive rate of the p-p38 protein and uPA protein expression in the breast cancer tissues was 56.7% and 60.0%,respectively. The expression of p-p38 was positively related to the expression of uPA （r = 0.316, P 〈 0.05）. The expression of p-p38 and uPA was related to lymph node metastasis and the TNM stage （P 〈 0.05）, but it was not related to the patient＇s age or tumor size （P 〉 0.05）. The expression of p-p38 and uPA in MDA- MB-231 cells was higher than that in MCF-7 cells. SB203580 inhibited the p38 MAPK pathway and reduced uPA protein expression.CONCLUSION The p38 MAPK-signaling pathway promotes breast cancer malignant progression by up-regulating uPA expression ,and it may be an important process in breast cancer invasion and metastasis.
文摘The matrix metalloproteinases (MMPs) are a family of zinc-dependent endopeptidases that play a key role in both normal and pathological processes involving tissue remodeling events. The expression of these proteolytic enzymes is highly regulated by a balance between extracellular matrix (ECM) deposition and its degradation, and is controlled by growth factors, cytokines, hormones, as well as interactions with the ECM macromolecules. Furthermore, the activity of the MMPs is regulated by their natural endogenous inhibitors, which are members of the tissue inhibitor of metalloproteinases (TIMP) family. In the normal mammary gland, MMPs are expressed during ductal development, lobulo-alveolar development in pregnancy and involution after lactation. Under pathological conditions, such as tumorigenesis, the dysregulated expression of MMPs play a role in tumor initiation, progression and malignant conversion as well as facilitating invasion and motastasis of malignant cells through degradation of the ECM and basement membranes. Matrix metalloproteinases and their expression in mammary gland
基金a grant from the National Natural Science Foundation of China (No. 30370553).
文摘OBJECTIVE To investigate the efficiency of gene silencing by CXCR4- siRNAs （small interfering RNA）, and to examine the invasive ability and the expression of other metastatic-associated genes in siRNA-treated breast cancer cells.METHODS Three siRNAs were designed and cloned into the pSilenc ^TM 3.1-H1 neo vector. The reconstructed plasmids were purified and transfected into the T47D breast cancer cell line, which highly expressed CXCR4. The amount of CXCR4 expression in the transfected cells was measured by flow cytometry and Real-time PCR. Cell invasive ability was evaluated using 24-well Matrigel invasion chambers. In addition, the expression of other metastatic-associated genes, such as E-cad, IGFBP-5, FN and MMP-2, was assessed by Real-time PCRRESULTS The suppression rates of CXCR4 mRNA expression reached 95.7%, 85.9% and 98.3%compared with control-siRNA cells in the 3 CXCR4- siRNA T47D cells respectively. FCM assays for CXCR4 protein expression showed a similar inhibitory effect. The invasion indexes of these CXCR4- siRNA cells were 0.037, 0.290 and 0.188 respectively compared with control- siRNA cells. After treatment of the cells with CXCR4-siRNA, the expression of E-cad showed an upward tendency and that of IGFBP-5 had a downward trend, while alteration in expression of FN and MMP2 varied without a consistant effect.CONCLUSION CXCR4 plays an important role in modulating migra- tion of human breast cancer cells. Small interfering RNA can significantly silence the CXCR4 gene in the human T47D breast cancer cell line. The results of this study strengthen the need for further research on novel gene therapy against breast cancer metastasis.
文摘OBJECTIVE To investigate the expression of the RECK gene in human breast （cancer） cell lines, and to determine the relationship between RECK gene expression and the invasive capacity of the breast cancer cell lines.METHODS The invasive capacity of breast （cancer） cell lines including HBL-100, MCF-7 and MDA-MB-435S were determined by the Tran- swell method. The protein expression levels of RECK, MMP-2 and MMP- 9 genes in these three cell lines were measured by immunocytochemical methods. The expressions of the RECK gene and protein level were measured hv RT-PCR and Westrn hints in the cell lines respectively.RESULTS The order of the nvasive capacity of the breast （cancer） cell lines was MDA-MB-435S, being the highest, and HBL-100, being the lowest. The invasive capacity difference between any two groups among the three groups was significant （P〈0.01）. The protein expression level of the RECK gene in the HBL-100 cell line was highest, and no expression was detected in MDA-MB-435S cells. Moreover, the expression of the RECK gene was negatively correlated with the expression of the MMP-2 and MMP-9 genes. The mRNA level of the RECK gene in HBL-100 cells was the highest, but no expression was found in the MDA-MB-435S cells （P〈0.001）.CONCLUSION There was a significant negative correlation between the expression level of the RECK gene and invasive capacity in vitro, and the RECK gene expression showed an inverse proportion to that of the MMP-2, MMP-9 genes.
文摘AIM： We examined quantitative mRNA expression of growth factor receptors （c-erbB-1, c-erbB-2） and the anti-apoptosis gene survivin known to be regulated in pancreatic adenocarcinomas and compared the expression pattern with that in carcinomas of the papilla of Vater. METHODS： Quantitative real-time reverse transcriptase- PCR （QRT-PCR, Taqman^TM） was performed to analyze mRNA expression levels of c-erbB-1, c-erbB-2 and survivin in normal and corresponding tumor samples of 31 pancreatic adenocarcinomas and 8 cancers of the papilla of Vater. RESULTS： The overall median mRNA expression of survivin was significantly increased in both adenocarcinoma of the pancreas （P〈0.01） and papilla of Vater （P〈0.008） compared with uninvolved normal control tissue. In pancreatic cancer, expression of c-erbB-1 was significantly decreased compared with the normal pancreatic tissue （P〈0.03）, whereas in the cancer of the papilla of Vater expression of c-erbB-2 was significantly downregulated （P〈0.05） compared with the paired normal samples. Gene expression was not associated with tumor stage, differentiation or prognosis. CONCLUSION： The common anti-apoptosis gene survivin is overexpressed both in the cancer of the papilla of Vater and pancreas. In contrast, the growth factor receptor genes c-erbB-1 and c-erbB-2 are differentially regulated in both tumor entities adding further evidence that pancreatic cancer is biologically different from the cancer of papilla of Vater.
文摘A malignant transformed mammary epithelial cell line (11A1) was transfected with liposome encapsulated eukaryotic expression plasmid pCMV-neo-RB, yielding 4 constant clones which have obvious pheno-typic reversion changes, and named 11A1-R1-R4 respectively. Further experiments showed that the 11A1-R1 behaved like normal epithelial cells in both morphological and biological characteristics, with decreased clonogenicity in solid argar medium as well as decreased tumorigenicity. Northern blot hybridization showed increased expression of RB gene and decreased expression of c-myc gene in 11A1-R1, 11A1-R2 cells compared to 11A1 cells. This was an ideal phenotypic reversion model for epithelial transformed cell line and demonstrated that the RB gene can reexpress and suppress malignant phenotype in RB inactive cells.
