为研究鞘脂激活蛋白原(Prosaposin)对细胞增殖、细胞凋亡的调控及其可能的分子机制,以pcDNA3.1 in NIH3T3阴性对照细胞株和过表达prosaposin的Psap-Myc in NIH3T3细胞株为模型,噻唑蓝(MTT)比色法检测prosaposin对细胞增殖的影响;Annexi...为研究鞘脂激活蛋白原(Prosaposin)对细胞增殖、细胞凋亡的调控及其可能的分子机制,以pcDNA3.1 in NIH3T3阴性对照细胞株和过表达prosaposin的Psap-Myc in NIH3T3细胞株为模型,噻唑蓝(MTT)比色法检测prosaposin对细胞增殖的影响;Annexin V联合碘化丙啶(Propidium iodide,PI)法检测血清饥饿状态下prosaposin对细胞凋亡的影响;Western blotting检测PI3K/Akt信号通路中蛋白磷酸化水平的变化;Real-time PCR检测PI3K/Akt信号通路下游靶分子表达水平的改变。结果表明prosaposin可活化PI3K/Akt信号通路,提高AktSer473的磷酸化水平,抑制细胞周期抑制基因P27KIP1的表达,上调细胞周期蛋白Cyclin D1的表达,促进细胞周期从G1→S期进展;诱导survival基因cIAP1、cIAP2的表达,促进细胞存活。这些结果提示,prosaposin对细胞增殖和凋亡的调控可能是通过PI3K/Akt信号通路及其下游靶分子进行的。展开更多
<abstract>The recent development of a prosaposin -/- mouse model has allowed the investigation of the role of prosaposin in the development of the male reproductive organs. A morphometric analysis of the male re...<abstract>The recent development of a prosaposin -/- mouse model has allowed the investigation of the role of prosaposin in the development of the male reproductive organs. A morphometric analysis of the male reproductive system of 37 days old mice revealed that prosaposin ablation produced a 30 % reduction in size and weight of the testes, 37 % of the epididymis, 75 % of the seminal vesicles and 60 % of the prostate glands. Light microscopy (LM) showed that smaller testis size from homozygous mutant mice was associated with reduced spermiogenesis. Both, dorsal and ventral lobules of the prostate glands were underdeveloped in the homozygous mutant. LM analysis also showed that prostatic alveoli were considerably smaller and lined by shorter epithelial cells in the homozygous mutant. Smaller tubular diameter and shorter undifferentiated epithelial cells were also observed in seminal vesicles and epididymis. In the efferent ducts of the homozygous mutant mice, the epithelium was composed exclusively of ciliated cells in contrast to the heterozygotes, which showed the presence of nonciliated cells. Radioimmunoassays demonstrated that testosterone levels were normal or higher in mice with the inactivated prosaposin gene. Immunostaining of prostate sections with an anti-androgen receptor antibody showed that the epithelial cells lining the alveoli express androgen receptor in both the heterozygous and homozygous tissue. Similarly, sections immunostained with antibodies to the phosphorylated MAPKs and Akts strongly reacted with tall prostatic secretory cells in prostate from heterozygous mouse. On the other hand, the epithelial cells in the homozygous prostate remained unstained or weakly stained. These findings demonstrate that inactivation of the prosaposin gene affected the development of the prostate gland and some components of the MAPK pathway. 63 )展开更多
Prosaposin(Psap)has multiple cellular functions.It is involved in the development of the reproductive system,nervous system,and prostate cancer as well as in the regulation of sphingolipid catabolism by activating sev...Prosaposin(Psap)has multiple cellular functions.It is involved in the development of the reproductive system,nervous system,and prostate cancer as well as in the regulation of sphingolipid catabolism by activating several lysosomal hydrolases involved in the metabolism of various sphingolipids.In this research,it was found to be a novel interaction partner for Rhox5 using yeast two-hybrid screening.The interaction between Rhox5 and the full-length prosapsoin(the transcript without exon 8)as well as the C-terminal domain of prosaposin,was further confirmed in both yeast two hybrid analysis and in vitro assay.It suggested that the C-terminal domain of prosaposin may be critical for the Rhox5-prosaposin interaction.Given the important roles played by both Rhox5 and prosaposin in maintaining the differentiation of male reproductive organs,spermatogenesis,and fertilization,the interaction between Rhox5 and prosaposin might regulate the development of male reproductive organs dynamically.展开更多
Objective To investigate the prosaposin （PSAP） expression in polycystic ovaries. Methods The expression of PSAP was examined using immunohistochemistry, quantitative RT-PCR and Western blotting in a rat model with l...Objective To investigate the prosaposin （PSAP） expression in polycystic ovaries. Methods The expression of PSAP was examined using immunohistochemistry, quantitative RT-PCR and Western blotting in a rat model with letrozole-induced polycystic ovary syndrom （PCOS）. Results Letrozole-induced PCOS rat models were similar to those in human PCOS in endocrine disturbances, metabolic characteristics and morphology features. And in these models both mRNA and protein levels of PSAP were significantly increased. Conclusion PSAP was overexpressed in the letrozole-induced PCOS rat model and might play an important role in the oceurance and development of PCOS.展开更多
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
Prosaposin is a multifunctional protein that is the precursor of four lysosomal activator proteins knownas saposin when located in the lysosome and is an integral membrane protein when secreted.Despite of its function...Prosaposin is a multifunctional protein that is the precursor of four lysosomal activator proteins knownas saposin when located in the lysosome and is an integral membrane protein when secreted.Despite of its function duringsphingolipid catabolism,prosaposin also plays an important role in reproductive system,nervous system and the develop-ment of prostate cancer.展开更多
文摘为研究鞘脂激活蛋白原(Prosaposin)对细胞增殖、细胞凋亡的调控及其可能的分子机制,以pcDNA3.1 in NIH3T3阴性对照细胞株和过表达prosaposin的Psap-Myc in NIH3T3细胞株为模型,噻唑蓝(MTT)比色法检测prosaposin对细胞增殖的影响;Annexin V联合碘化丙啶(Propidium iodide,PI)法检测血清饥饿状态下prosaposin对细胞凋亡的影响;Western blotting检测PI3K/Akt信号通路中蛋白磷酸化水平的变化;Real-time PCR检测PI3K/Akt信号通路下游靶分子表达水平的改变。结果表明prosaposin可活化PI3K/Akt信号通路,提高AktSer473的磷酸化水平,抑制细胞周期抑制基因P27KIP1的表达,上调细胞周期蛋白Cyclin D1的表达,促进细胞周期从G1→S期进展;诱导survival基因cIAP1、cIAP2的表达,促进细胞存活。这些结果提示,prosaposin对细胞增殖和凋亡的调控可能是通过PI3K/Akt信号通路及其下游靶分子进行的。
文摘<abstract>The recent development of a prosaposin -/- mouse model has allowed the investigation of the role of prosaposin in the development of the male reproductive organs. A morphometric analysis of the male reproductive system of 37 days old mice revealed that prosaposin ablation produced a 30 % reduction in size and weight of the testes, 37 % of the epididymis, 75 % of the seminal vesicles and 60 % of the prostate glands. Light microscopy (LM) showed that smaller testis size from homozygous mutant mice was associated with reduced spermiogenesis. Both, dorsal and ventral lobules of the prostate glands were underdeveloped in the homozygous mutant. LM analysis also showed that prostatic alveoli were considerably smaller and lined by shorter epithelial cells in the homozygous mutant. Smaller tubular diameter and shorter undifferentiated epithelial cells were also observed in seminal vesicles and epididymis. In the efferent ducts of the homozygous mutant mice, the epithelium was composed exclusively of ciliated cells in contrast to the heterozygotes, which showed the presence of nonciliated cells. Radioimmunoassays demonstrated that testosterone levels were normal or higher in mice with the inactivated prosaposin gene. Immunostaining of prostate sections with an anti-androgen receptor antibody showed that the epithelial cells lining the alveoli express androgen receptor in both the heterozygous and homozygous tissue. Similarly, sections immunostained with antibodies to the phosphorylated MAPKs and Akts strongly reacted with tall prostatic secretory cells in prostate from heterozygous mouse. On the other hand, the epithelial cells in the homozygous prostate remained unstained or weakly stained. These findings demonstrate that inactivation of the prosaposin gene affected the development of the prostate gland and some components of the MAPK pathway. 63 )
基金supported by the National Natural Sciences Foundation of China(No.39770411)Doctoral Fund of Ministry of Education of China(No.20060559006).
文摘Prosaposin(Psap)has multiple cellular functions.It is involved in the development of the reproductive system,nervous system,and prostate cancer as well as in the regulation of sphingolipid catabolism by activating several lysosomal hydrolases involved in the metabolism of various sphingolipids.In this research,it was found to be a novel interaction partner for Rhox5 using yeast two-hybrid screening.The interaction between Rhox5 and the full-length prosapsoin(the transcript without exon 8)as well as the C-terminal domain of prosaposin,was further confirmed in both yeast two hybrid analysis and in vitro assay.It suggested that the C-terminal domain of prosaposin may be critical for the Rhox5-prosaposin interaction.Given the important roles played by both Rhox5 and prosaposin in maintaining the differentiation of male reproductive organs,spermatogenesis,and fertilization,the interaction between Rhox5 and prosaposin might regulate the development of male reproductive organs dynamically.
基金a Colleges and Universities Scientific Program supported by Education Department of Liaoning Province (LT2010106)
文摘Objective To investigate the prosaposin （PSAP） expression in polycystic ovaries. Methods The expression of PSAP was examined using immunohistochemistry, quantitative RT-PCR and Western blotting in a rat model with letrozole-induced polycystic ovary syndrom （PCOS）. Results Letrozole-induced PCOS rat models were similar to those in human PCOS in endocrine disturbances, metabolic characteristics and morphology features. And in these models both mRNA and protein levels of PSAP were significantly increased. Conclusion PSAP was overexpressed in the letrozole-induced PCOS rat model and might play an important role in the oceurance and development of PCOS.
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.
文摘Prosaposin is a multifunctional protein that is the precursor of four lysosomal activator proteins knownas saposin when located in the lysosome and is an integral membrane protein when secreted.Despite of its function duringsphingolipid catabolism,prosaposin also plays an important role in reproductive system,nervous system and the develop-ment of prostate cancer.
