Aim:To determine the effect of saposin C (a known trophic domain of prosaposin) on proliferation,migration and invasion,as well as its effect on the expression of urokinase plasmonogen activator (uPA),its receptor (uP...Aim:To determine the effect of saposin C (a known trophic domain of prosaposin) on proliferation,migration and invasion,as well as its effect on the expression of urokinase plasmonogen activator (uPA),its receptor (uPAR) and matrix metalloproteinases (MMP)-2 and -9 in normal and malignant prostate cells.In addition,we tested whether saposin C can activate p42/44 and stress-activated protein kinase/c-Jun NH_2-terminal kinase (SAPK/JNK) signal transduction pathways of the mitogen-activated protein kinase (MAPK) superfamily.Methods:We employed West- ern blot analysis,phospho-specific antibodies,cell proliferation assay,reverse transcriptase-polymerase chain reaction, in vitro kinase assays and migration and invasion to determine the effect of saposin C on various biological behaviors of prostate stromal and cancer cells.Results:Saposin C,in a cell type-specific manner,upregulates uPA/uPAR and immediate early gene c-Jun expression,stimulates cell proliferation,migration and invasion and activates p42/44 and SAPK/JNK MAPK pathways in prostate stromal and cancer cells.Normal prostate epithelial cells were not responsive to saposin C treatment in the above studies.Conclusion:Saposin C functions as a multipotential modulator of diverse biological activities in prostate cancer and stromal cells.These results strongly suggest that saposin C functions as a potent growth factor for prostatic cells and may contribute to prostate carcinogenesis and/or the development of hormone-refractory prostate cancer.展开更多
Aim： To determine the effects of the functional domain of saposin C （neurotrophic peptide [NP]） on androgen receptor （AR） expression and transcriptional activity. Methods： We constructed DNA vectors expressing N...Aim： To determine the effects of the functional domain of saposin C （neurotrophic peptide [NP]） on androgen receptor （AR） expression and transcriptional activity. Methods： We constructed DNA vectors expressing NP or a chimeric peptide of the viral TAT transduction domain and NP （TAT-NP） using gene cloning technology. The effects of ectopic expression of NP or TAT-NP on cell growth were examined by 3-（4, 5-dimethylthiazol-2-yl）-2, 5-diphenyl-2H-tetrazolium bromide （MTT） assay. Reverse transcription-polymerase chain reaction （RT-PCR）, Western blot, transient transfection and reporter gene assays were used to determine the effects of NP on AR expression and activation. Results： NP stimulated proliferation of androgen responsive LNCaP cells in the absence of androgens. RT-PCR and Western blot analyses showed that ectopic expression of NP resulted in induction of AR gene expression, and that the NP-stimulated expression of AR could be synergistically enhanced in the presence of androgens. Furthermore, reporter gene assay results showed that NP could enhance AR transactivation by increasing androgen-inducible gene reporter activity. Conclusion： We provided evidence that ectopic expression of saposin C-originated NP could upregulate AR gene expression and activate the AR transcriptional function in an androgen-independent manner in prostate cancer cells.展开更多
Pancreatic cancer is one of the deadliest of cancers with a five-year survival of roughly 8%.Current therapies are:surgery,radiation and chemotherapy.Surgery is curative only if the cancer is caught very early,which i...Pancreatic cancer is one of the deadliest of cancers with a five-year survival of roughly 8%.Current therapies are:surgery,radiation and chemotherapy.Surgery is curative only if the cancer is caught very early,which is rare,and the latter two modalities are only marginally effective and have significant side effects.We have developed a nanosome comprised of the lysosomal protein,saposin C(SapC)and the acidic phospholipid,dioleoylphosphatidylserine(DOPS).In the acidic tumor microenvironment,this molecule,SapC-DOPS,targets the phosphatidylserine cancer-biomarker which is predominantly elevated on the surface of cancer cells.Importantly,SapC-DOPS can selectively target pancreatic tumors and metastases.Furthermore,SapC-DOPS has exhibited an impressive safety profile with only a few minor side effects in both preclinical experiments and in phase I clinical trials.With the dismal outcomes for pancreatic cancer there is an urgent need for better treatments and SapC-DOPS is a good candidate for addition to the oncologist’s toolbox.展开更多
Granulysin is a cytotoxic granular protein that was identified from human T cells by using the gene subtraction method in 1987. Based on its amino acid homology, granulysin belongs to the saposin-like protein family. ...Granulysin is a cytotoxic granular protein that was identified from human T cells by using the gene subtraction method in 1987. Based on its amino acid homology, granulysin belongs to the saposin-like protein family. The bioactive 9-k Da form of granulysin is processed from the 15-k Da pro-product in the cytoplasmic granules. It is expressed in CD8-positive αβT cells 5 d after mitogenic stimulation and constitutively in natural killer(NK) cells and γδT cells, although regulation of its expression has not yet been precisely determined. The 9-k Da granulysin form has anti-microbial activity against microorganisms such as bacteria, fungi, mycobacteria and parasites, as well as tumoricidal activity against some tumors at 1-10 μmol/L concentrations. Granulysin is secreted in both Ca-dependent and-inde-pendent manners. In sera, only the 15-k Da form is detectable and is expected to be a biomarker for immune potency, acute viral infection, anti-tumor immune reaction, acute graft vs host disease, and NK cell associated neoplasm.展开更多
文摘Aim:To determine the effect of saposin C (a known trophic domain of prosaposin) on proliferation,migration and invasion,as well as its effect on the expression of urokinase plasmonogen activator (uPA),its receptor (uPAR) and matrix metalloproteinases (MMP)-2 and -9 in normal and malignant prostate cells.In addition,we tested whether saposin C can activate p42/44 and stress-activated protein kinase/c-Jun NH_2-terminal kinase (SAPK/JNK) signal transduction pathways of the mitogen-activated protein kinase (MAPK) superfamily.Methods:We employed West- ern blot analysis,phospho-specific antibodies,cell proliferation assay,reverse transcriptase-polymerase chain reaction, in vitro kinase assays and migration and invasion to determine the effect of saposin C on various biological behaviors of prostate stromal and cancer cells.Results:Saposin C,in a cell type-specific manner,upregulates uPA/uPAR and immediate early gene c-Jun expression,stimulates cell proliferation,migration and invasion and activates p42/44 and SAPK/JNK MAPK pathways in prostate stromal and cancer cells.Normal prostate epithelial cells were not responsive to saposin C treatment in the above studies.Conclusion:Saposin C functions as a multipotential modulator of diverse biological activities in prostate cancer and stromal cells.These results strongly suggest that saposin C functions as a potent growth factor for prostatic cells and may contribute to prostate carcinogenesis and/or the development of hormone-refractory prostate cancer.
基金Supported by National Natural Science Foundation of China(No.31101266)Scientific Research Program Funded by Education Department of Shaanxi Province(No.12JK0711)~~
文摘Aim： To determine the effects of the functional domain of saposin C （neurotrophic peptide [NP]） on androgen receptor （AR） expression and transcriptional activity. Methods： We constructed DNA vectors expressing NP or a chimeric peptide of the viral TAT transduction domain and NP （TAT-NP） using gene cloning technology. The effects of ectopic expression of NP or TAT-NP on cell growth were examined by 3-（4, 5-dimethylthiazol-2-yl）-2, 5-diphenyl-2H-tetrazolium bromide （MTT） assay. Reverse transcription-polymerase chain reaction （RT-PCR）, Western blot, transient transfection and reporter gene assays were used to determine the effects of NP on AR expression and activation. Results： NP stimulated proliferation of androgen responsive LNCaP cells in the absence of androgens. RT-PCR and Western blot analyses showed that ectopic expression of NP resulted in induction of AR gene expression, and that the NP-stimulated expression of AR could be synergistically enhanced in the presence of androgens. Furthermore, reporter gene assay results showed that NP could enhance AR transactivation by increasing androgen-inducible gene reporter activity. Conclusion： We provided evidence that ectopic expression of saposin C-originated NP could upregulate AR gene expression and activate the AR transcriptional function in an androgen-independent manner in prostate cancer cells.
基金Supported by Pancreatic Cancer Action Network Translational Research Grant,No.20-65-QIXIGive Hope Foundation,National Institutes of Health,No.R01CA158372 and No.R21NS095047CCTST Pilot Collaborative Studies,Bearcats Against Cancer,and Hematology-Oncology Programmatic Support from University of Cincinnati College of Medicine(to Qi X).
文摘Pancreatic cancer is one of the deadliest of cancers with a five-year survival of roughly 8%.Current therapies are:surgery,radiation and chemotherapy.Surgery is curative only if the cancer is caught very early,which is rare,and the latter two modalities are only marginally effective and have significant side effects.We have developed a nanosome comprised of the lysosomal protein,saposin C(SapC)and the acidic phospholipid,dioleoylphosphatidylserine(DOPS).In the acidic tumor microenvironment,this molecule,SapC-DOPS,targets the phosphatidylserine cancer-biomarker which is predominantly elevated on the surface of cancer cells.Importantly,SapC-DOPS can selectively target pancreatic tumors and metastases.Furthermore,SapC-DOPS has exhibited an impressive safety profile with only a few minor side effects in both preclinical experiments and in phase I clinical trials.With the dismal outcomes for pancreatic cancer there is an urgent need for better treatments and SapC-DOPS is a good candidate for addition to the oncologist’s toolbox.
基金Supported by The Grant-in-Aid for Scientific Research from Ministry of Education,Science and Culture Japan,24591541 to Nagasawa M
文摘Granulysin is a cytotoxic granular protein that was identified from human T cells by using the gene subtraction method in 1987. Based on its amino acid homology, granulysin belongs to the saposin-like protein family. The bioactive 9-k Da form of granulysin is processed from the 15-k Da pro-product in the cytoplasmic granules. It is expressed in CD8-positive αβT cells 5 d after mitogenic stimulation and constitutively in natural killer(NK) cells and γδT cells, although regulation of its expression has not yet been precisely determined. The 9-k Da granulysin form has anti-microbial activity against microorganisms such as bacteria, fungi, mycobacteria and parasites, as well as tumoricidal activity against some tumors at 1-10 μmol/L concentrations. Granulysin is secreted in both Ca-dependent and-inde-pendent manners. In sera, only the 15-k Da form is detectable and is expected to be a biomarker for immune potency, acute viral infection, anti-tumor immune reaction, acute graft vs host disease, and NK cell associated neoplasm.
