AIM:To determine the expression and function of epithelial membrane protein 1(EMP1)in colorectal carcinoma.METHODS:Colorectal samples were taken from cancer lesions and adjacent normal tissue in colorectal cancer pati...AIM:To determine the expression and function of epithelial membrane protein 1(EMP1)in colorectal carcinoma.METHODS:Colorectal samples were taken from cancer lesions and adjacent normal tissue in colorectal cancer patients immediately after endoscopic biopsy.A portion of the sample was either fixed in 4%paraformaldehyde and embedded in paraffin for immunohistochemistry or stored in liquid nitrogen for Western blot.In order to determine protein expression of EMP1in colorectal cancer(n=63)and normal tissue(n=31),semi-quantitative immunohistochemistry and Western blot were utilized.For in vitro studies,the human colorectal cancer cell line SW-480 was maintained in RPMI-1640 medium supplemented with 10%fetal bovine serum.Recombinant lentivirus mediated overexpression of EMP1 in SW-480 cells was quantified by real-time reverse transcription-polymerase chain reaction and Western blot.Control SW-480 cells were transfected with an empty vector.To further study the effect of EMP1 overexpression in SW-480 cells,cell proliferation,apoptosis,migration and invasion assays were conducted.RESULTS:Expression of EMP1 was significantly lower in colorectal cancer tissue than in normal tissue using both immunohistochemistry(39.7%vs 90.3%of tissues,P<0.05)and Western blot(0.126±0.022 vs0.632±0.053,P<0.05).The level of EMP1 protein expression was not correlated with gender,age,or tumor location.Decreased expression of EMP1 was significantly correlated with T stage,lymph node metastasis,clinic stage,and histological grade in patients with colorectal cancer(P<0.05).According to Kaplan-Meier analysis,low EMP1 expression correlated significantly with poor overall five-year survival(34.2%vs 64.0%survival,P<0.05).SW-480 cells transfected with EMP1 had a lower survival fraction,higher cell apoptosis(12.1%±1.3%vs 3.1%±0.6%,P<0.05),a significant decrease in migration and invasion(124.0±17.0 and 87.0±12.0,respectively vs 213.0±29.0 and 178.0±21.0,respectively,P<0.05),higher caspase-9(0.635±0.063 vs0.315±0.032,P<0.05),and lower VEGFC protein expression(0.229±0.021 vs 0.519±0.055,P<0.05)relative to cells not transfected with EMP1.CONCLUSION:Low EMP1 expression in colorectal cancer is associated with increased disease severity,suggesting that EMP1 may be a negative regulator of colorectal cancer.展开更多
Here we report a family with a clinical spectrum of Pachyonychia Congenita Tarda (PCT) encompassing two generations via a balanced chromosomal translocation between 4q26 and 12p12.3. We discuss the effects of chromoso...Here we report a family with a clinical spectrum of Pachyonychia Congenita Tarda (PCT) encompassing two generations via a balanced chromosomal translocation between 4q26 and 12p12.3. We discuss the effects of chromosomal translocations on gene expression through involved breakpoints and structural gene abnormalities detected by array CGH. We believe that the family we present gives further insight to the better understanding of molecular and structural basis of keratin disorders, and to the late onset and genetic basis of PCT through the possible role of C-type lectins and human epithelial membrane protein1 (EMP1). Better understanding of the molecular basis of keratin disorders is the foundation for improved diagnosis, genetic counseling and novel therapeutic approaches to overcome the current treatment limitations related to this disease.展开更多
Aim： To investigate the possible functions of human sperm membrane protein （hSMP-1） in the process of fertilization. Methods： A 576-bp cDNA fragment of HSD-1 gene coding for the extracellular domain of hSMP-1 was ...Aim： To investigate the possible functions of human sperm membrane protein （hSMP-1） in the process of fertilization. Methods： A 576-bp cDNA fragment of HSD-1 gene coding for the extracellular domain of hSMP-1 was cloned and expressed. The localization of this protein on human and mouse sperm was determined by indirect immunofluorescent staining by using anti-recombinant hSMP-1 （anti-rhSMP-1） antibodies. Sperm acrosome reaction and spermzona pellucida （ZP） binding assay were carried out in 10-week-old BALB/c mice. Results： Recombinant hSMP-1 was successfully cloned and expressed. The expression of the native protein was limited on the acrosome of human and mouse sperm. Treatment of anti-rhSMP-1 antibodies significantly decreased the average number of sperms bound to each egg. Meanwhile, the percentage of acrosome reaction was decreased in comparison to pre-immune control after treatment with anti-rhSMP-1 （P 〈 0.05）. Conclusion： The results suggest that anti-rhSMP-1 antibody inhibited mouse acrosome reaction and sperm-ZP binding.展开更多
Objective： To investigate the expressions of beta-catenin, protein APC （adenomatous polyposis coil protein）, c-myc and cyclin D1 and their implication in ovarian epithelial tumor. Methods： Immunohistochemical stai...Objective： To investigate the expressions of beta-catenin, protein APC （adenomatous polyposis coil protein）, c-myc and cyclin D1 and their implication in ovarian epithelial tumor. Methods： Immunohistochemical staining with SP method was conducted to identify the expressions of beta-catenin, APC protein, c-myc and cyclin D1 in ovarian epithelial tumor in 48 cases. Results： The abnormal expression rate of beta-catenin in malignant and borderline ovarian epithelial tumors was higher than that in benign epithelial tumors （P〈0.01）. The expression rates of c-myc and cyclin-D1 in ovarian malignant and borderline epithelial tumors were higher than those in benign epithelial tumors too（P〈0.05）. The prevalence of APC protein positive expression in benign epithelial tumors were significantly greater than that in malignant epithelial tumors （P〈0.05）. A significant negative correlation was found between beta-catenin and APC protein in ovarian epithelial tumors; while a significant positive correlation was found between beta-catenin, c-myc and cyclin-D1 in ovarian epithelial tumor （P〈0.05）. Conclusion： The abnormal expressions of Beta-catenin, APC protein, c-myc and cyclin-D1 might be used to indicate the malignance transform of ovarian epithelial tumors.展开更多
AIM: To investigate the role of nuclear factor κB(NF-κB) in the regulation of Epstein-Barr virus(EBV) latent membrane protein 1(LMP1) in EBV transformed cells. METHODS: LMP1 expression was examined in EBV transforme...AIM: To investigate the role of nuclear factor κB(NF-κB) in the regulation of Epstein-Barr virus(EBV) latent membrane protein 1(LMP1) in EBV transformed cells. METHODS: LMP1 expression was examined in EBV transformed human B lymphocytes with modulation of NF-κB activity. RESULTS: EBV infection is associated with several human cancers. EBV LMP1 is required for efficient transformation of adult primary B cells in vitro, and is expressed in several pathogenic stages of EBVassociated cancers. Regulation of EBV LMP1 involves both viral and cellular factors. LMP1 activates NF-κB signaling pathway that is a part of the EBV transformation program. However, the relation between NF-κB and LMP1 expression is not well established yet. In this report, we found that blocking the NF-κB activity by Inhibitor of κB stimulated LMP1 expression, while the overexpression of NF-κB repressed LMP1 expression in EBV-transformed IB4 cells. In addition, LMP1 repressed its own promoter activities in reporter assays, and the repression was associated with the activation of NF-κB. Moreover, NF-κB alone is sufficient to repress LMP1 promoter activities. CONCLUSION: Our data suggest LMP1 may repress its own expression through NF-κB in EBV transformed cells and shed a light on LMP1 regulation during EBV transformation.展开更多
文摘AIM:To determine the expression and function of epithelial membrane protein 1(EMP1)in colorectal carcinoma.METHODS:Colorectal samples were taken from cancer lesions and adjacent normal tissue in colorectal cancer patients immediately after endoscopic biopsy.A portion of the sample was either fixed in 4%paraformaldehyde and embedded in paraffin for immunohistochemistry or stored in liquid nitrogen for Western blot.In order to determine protein expression of EMP1in colorectal cancer(n=63)and normal tissue(n=31),semi-quantitative immunohistochemistry and Western blot were utilized.For in vitro studies,the human colorectal cancer cell line SW-480 was maintained in RPMI-1640 medium supplemented with 10%fetal bovine serum.Recombinant lentivirus mediated overexpression of EMP1 in SW-480 cells was quantified by real-time reverse transcription-polymerase chain reaction and Western blot.Control SW-480 cells were transfected with an empty vector.To further study the effect of EMP1 overexpression in SW-480 cells,cell proliferation,apoptosis,migration and invasion assays were conducted.RESULTS:Expression of EMP1 was significantly lower in colorectal cancer tissue than in normal tissue using both immunohistochemistry(39.7%vs 90.3%of tissues,P<0.05)and Western blot(0.126±0.022 vs0.632±0.053,P<0.05).The level of EMP1 protein expression was not correlated with gender,age,or tumor location.Decreased expression of EMP1 was significantly correlated with T stage,lymph node metastasis,clinic stage,and histological grade in patients with colorectal cancer(P<0.05).According to Kaplan-Meier analysis,low EMP1 expression correlated significantly with poor overall five-year survival(34.2%vs 64.0%survival,P<0.05).SW-480 cells transfected with EMP1 had a lower survival fraction,higher cell apoptosis(12.1%±1.3%vs 3.1%±0.6%,P<0.05),a significant decrease in migration and invasion(124.0±17.0 and 87.0±12.0,respectively vs 213.0±29.0 and 178.0±21.0,respectively,P<0.05),higher caspase-9(0.635±0.063 vs0.315±0.032,P<0.05),and lower VEGFC protein expression(0.229±0.021 vs 0.519±0.055,P<0.05)relative to cells not transfected with EMP1.CONCLUSION:Low EMP1 expression in colorectal cancer is associated with increased disease severity,suggesting that EMP1 may be a negative regulator of colorectal cancer.
文摘Here we report a family with a clinical spectrum of Pachyonychia Congenita Tarda (PCT) encompassing two generations via a balanced chromosomal translocation between 4q26 and 12p12.3. We discuss the effects of chromosomal translocations on gene expression through involved breakpoints and structural gene abnormalities detected by array CGH. We believe that the family we present gives further insight to the better understanding of molecular and structural basis of keratin disorders, and to the late onset and genetic basis of PCT through the possible role of C-type lectins and human epithelial membrane protein1 (EMP1). Better understanding of the molecular basis of keratin disorders is the foundation for improved diagnosis, genetic counseling and novel therapeutic approaches to overcome the current treatment limitations related to this disease.
文摘Aim： To investigate the possible functions of human sperm membrane protein （hSMP-1） in the process of fertilization. Methods： A 576-bp cDNA fragment of HSD-1 gene coding for the extracellular domain of hSMP-1 was cloned and expressed. The localization of this protein on human and mouse sperm was determined by indirect immunofluorescent staining by using anti-recombinant hSMP-1 （anti-rhSMP-1） antibodies. Sperm acrosome reaction and spermzona pellucida （ZP） binding assay were carried out in 10-week-old BALB/c mice. Results： Recombinant hSMP-1 was successfully cloned and expressed. The expression of the native protein was limited on the acrosome of human and mouse sperm. Treatment of anti-rhSMP-1 antibodies significantly decreased the average number of sperms bound to each egg. Meanwhile, the percentage of acrosome reaction was decreased in comparison to pre-immune control after treatment with anti-rhSMP-1 （P 〈 0.05）. Conclusion： The results suggest that anti-rhSMP-1 antibody inhibited mouse acrosome reaction and sperm-ZP binding.
基金the Scientific Research Start Found of Chongqing Medical University(QD 200201) project of Chongqing Science and Technology Committee (No. 040307)
文摘Objective： To investigate the expressions of beta-catenin, protein APC （adenomatous polyposis coil protein）, c-myc and cyclin D1 and their implication in ovarian epithelial tumor. Methods： Immunohistochemical staining with SP method was conducted to identify the expressions of beta-catenin, APC protein, c-myc and cyclin D1 in ovarian epithelial tumor in 48 cases. Results： The abnormal expression rate of beta-catenin in malignant and borderline ovarian epithelial tumors was higher than that in benign epithelial tumors （P〈0.01）. The expression rates of c-myc and cyclin-D1 in ovarian malignant and borderline epithelial tumors were higher than those in benign epithelial tumors too（P〈0.05）. The prevalence of APC protein positive expression in benign epithelial tumors were significantly greater than that in malignant epithelial tumors （P〈0.05）. A significant negative correlation was found between beta-catenin and APC protein in ovarian epithelial tumors; while a significant positive correlation was found between beta-catenin, c-myc and cyclin-D1 in ovarian epithelial tumor （P〈0.05）. Conclusion： The abnormal expressions of Beta-catenin, APC protein, c-myc and cyclin-D1 might be used to indicate the malignance transform of ovarian epithelial tumors.
基金Supported by Grants from the NIH CA138213,RR15635Department of Defense W81XWH-12-1-0225(Luwen Zhang)Qianli Wang was partially supported by Undergraduate Creative Activities and Research Experiences and Beckman Scholars Program
文摘AIM: To investigate the role of nuclear factor κB(NF-κB) in the regulation of Epstein-Barr virus(EBV) latent membrane protein 1(LMP1) in EBV transformed cells. METHODS: LMP1 expression was examined in EBV transformed human B lymphocytes with modulation of NF-κB activity. RESULTS: EBV infection is associated with several human cancers. EBV LMP1 is required for efficient transformation of adult primary B cells in vitro, and is expressed in several pathogenic stages of EBVassociated cancers. Regulation of EBV LMP1 involves both viral and cellular factors. LMP1 activates NF-κB signaling pathway that is a part of the EBV transformation program. However, the relation between NF-κB and LMP1 expression is not well established yet. In this report, we found that blocking the NF-κB activity by Inhibitor of κB stimulated LMP1 expression, while the overexpression of NF-κB repressed LMP1 expression in EBV-transformed IB4 cells. In addition, LMP1 repressed its own promoter activities in reporter assays, and the repression was associated with the activation of NF-κB. Moreover, NF-κB alone is sufficient to repress LMP1 promoter activities. CONCLUSION: Our data suggest LMP1 may repress its own expression through NF-κB in EBV transformed cells and shed a light on LMP1 regulation during EBV transformation.
