Follistatin(FST) is an important regulator of skeletal muscle growth and adipose deposition through its ability to bind to several members of the transforming growth factor-β(TGF-β) superfamily, and thus may be a go...Follistatin(FST) is an important regulator of skeletal muscle growth and adipose deposition through its ability to bind to several members of the transforming growth factor-β(TGF-β) superfamily, and thus may be a good candidate for future animal breeding programs. However, the molecular mechanisms underlying the phenotypic changes have yet to be clarified in pig. We generated transgenic(TG) pigs that express human FST specifically in skeletal muscle tissues and characterized the phenotypic changes compared with the same tissues in wild-type pigs. The TG pigs showed increased skeletal muscle growth, decreased adipose deposition, and improved metabolism status(P<0.05). Transcriptome analysis detected important roles of the PIK3–AKT signaling pathway, calcium-mediated signaling pathway, and amino acid metabolism pathway in FST-induced skeletal muscle hypertrophy, and depot-specific oxidative metabolism changes in psoas major muscle. Furthermore, the lipid metabolism-related process was changed in adipose tissue in the TG pigs. Gene set enrichment analysis revealed that genes related to lipid synthesis, lipid catabolism, and lipid storage were down-regulated(P<0.01) in the TG pigs for subcutaneous fat, whereas genes related to lipid catabolism were significantly up-regulated(P<0.05) in the TG pigs for retroperitoneal fat compared with their expression levels in wild-type pigs. In liver, genes related to the TGF-β signaling pathway were over-represented in the TG pigs, which is consistent with the inhibitory role of FST in regulating TGF-β signaling. Together, these results provide new insights into the molecular mechanisms underlying the phenotypic changes in pig.展开更多
Follistatin （FS） is a secreted protein, which was originally isolated from porcine follicular fluid. Expression of follistatin is tightly regulated during porcine growth and development. To study the essential trans...Follistatin （FS） is a secreted protein, which was originally isolated from porcine follicular fluid. Expression of follistatin is tightly regulated during porcine growth and development. To study the essential transcriptional regions of the porcine FS promoter, ten primer pairs were designed to amplify segments with different lengths of the FS promoter from -1 800 to ＋16 bp. The products were then inserted into the pGL3-basic vector to analyze the relative luciferase activity. The results showed that the most remarkable changes of promoter activity were observed between constructs （-302/＋16 bp）-FS and （-180/＋16 bp）-FS （P〈0.01）. Further research showed that the reconstructed reporter plasmid lacking myeloid zinc finger 1 （MZF1） binding sequence had significantly decreased luciferase activity （P〈0.05）. Furthermore, the FS protein expression was significantly increased in PK15 cells while the MZF1 was overexpressed, suggesting that the short sequence ＂TCCCCACC＂ （the recognition site of transcription factor MZF1） was the most important for FS transcription activation in the porcine.展开更多
Objective:Follistatin(FST)inhibits the action of activin by interfering with the binding of activin to its receptor.Although the prognostic value of FST in various cancers has been investigated previously,studies rare...Objective:Follistatin(FST)inhibits the action of activin by interfering with the binding of activin to its receptor.Although the prognostic value of FST in various cancers has been investigated previously,studies rarely focused on hypopharyngeal carcinoma(HPC).In our study,the effect of FST expression on HPC tissues and cell lines was investigated.Methods:A total of 60 patients with HPC were recruited for this study.Levels of FST mRNA and protein were measured by quantitative polymerase chain reaction(PCR)and immunohistochemistry in HPC tissue samples and by qPCR in the HPC FaDu cells,as well as immortal nasopharyngeal epithelial cell line NP-69 cells.After silencing the FST expression in FaDu cells using lentivirus-mediated siRNA that was specific for FST mRNA,cell proliferation was determined by a Celigo assay.Tumor growth was monitored in nude mice and viability was determined by a methylthiazoletetrazolium assay.The ratio of cell cycle arrest and apoptosis was evaluated by flow cytometry.The colony formation ability was performed using Giemsa staining.In addition,wound healing and Transwell migration and invasion assays were performed for the analysis of cell motility.Results:FST expression was significantly higher in human HPC tissue and FaDu cells than in normal tissue and NP-69 cells.A higher expression of FST in HPC samples was positively correlated with advanced tumors.Moreover,FST knockdown by shRNA significantly decreased cell growth,colony formation,migration and invasion.Furthermore,FST silencing increased the cell apoptosis percentage,and arrested cell cycle in the S phase in FaDu cells.In addition,FST silencing suppressed tumor growth in vivo.Conclusions:Our results indicated that the FST gene was associated with HPC progression and may serve as a potential therapeutic target for the treatment of HPC.展开更多
Objective: To investigate the effects of follistatin(rhFS-288) on biosynthesis andsecretion of testosterone in rat Leydig cell in vitro.Methods: Leydig cells were isolated from Wistar rat testes by a discontinuous Per...Objective: To investigate the effects of follistatin(rhFS-288) on biosynthesis andsecretion of testosterone in rat Leydig cell in vitro.Methods: Leydig cells were isolated from Wistar rat testes by a discontinuous Per-coll gradient procedure. Purified cells were incubated in 24-well plate(105 cell/ml/well)and maintained for 24 h in a CO2 incubator. rhFS-288 and Ca2+ were added to the wellsindependently or jointly in both baseline (without hCG) and stimulation condition (1.0IU/ml of hCG) to observe the change of testosterone concentration in the media.Results: rhFS-288 showed a dose-dependent inhibiting effect on testosterone releasein baseline and stimulating condition. Ca2+ presented inhibitory effect either. Whereas,escape phenomenon emerged while Ca2+ concentration reached to 100 mmol/L. A com-bination of rhFS-288 with Ca2+ displayed a dose-dependent inhibition on testosterone se-cretion.Conclusion: rhFS-288 inhibits testosterone secretion in a dose-dependent manner.Calcium is thought to be the second messenger of FS action. The mechanism of escapephenomenon during high dose of Ca2+ along is unknown.展开更多
Molecular biomarkers are commonly used for the management of several types of malignant tumours in routine clinical practice. However, this is not the case for malignant gliomas. Cytokines and Angiogenesis factors are...Molecular biomarkers are commonly used for the management of several types of malignant tumours in routine clinical practice. However, this is not the case for malignant gliomas. Cytokines and Angiogenesis factors are potential candidates due to their intrinsic role in tumourigenesis. Pre- and post-operative serum from 36 malignant glioma patients and 36 controls was analysed using the Bio-Plex Pro Angiogenesis and Cytokines Assay (Bio-Rad, USA). Amongst the molecules tested, the serum concentration of follistatin was significantly higher in patients than in controls. Moreover, the serum concentration of follistatin of the patients postoperatively was significantly reduced compared to that preoperatively. Factors such as age and gender did not affect the concentrations of follistatin measured in the serum of patients pre- and post-operatively as well as healthy controls. This is the first report of follistatin as potential biomarker for the detection of malignant gliomas.展开更多
Follistatin(FST) is a monomeric glycoprotein highly enriched in cysteines and belongs to TGF-β superfamily. FST can suppress the secretion of follicle-stimulating hormone and plays a vital role in the reproduction of...Follistatin(FST) is a monomeric glycoprotein highly enriched in cysteines and belongs to TGF-β superfamily. FST can suppress the secretion of follicle-stimulating hormone and plays a vital role in the reproduction of vertebrates. We used rapid amplification of cDNA ends technology to clone the FST gene of half-smooth tongue sole,Cynoglossus semilaevis. We characterized its phylogenetic context and expression patterns to elucidate its function in the breeding season. The full-length sequence of FST is 1 455 bp and encodes a protein of 321 amino acids. We investigated the expression pattern of FST in C. semilaevis at different stages of reproduction using reverse transcription-polymerase chain reaction(RTPCR). FST m RNA was expressed in all 13 tissues analyzed,and was expressed at high levels in gonad and at slightly lower levels in gill and brain. During the reproductive cycle of C. semilaevis,the transcript level of FST was the highest in the perinucleolus stage,decreased in the primary yolk stage,slightly increased in the tertiary yolk stage,and then reduced to a minimal level in the atretic follicles stage of the ovary. We concluded that FST suppressed follicle-stimulating hormone,which stimulated oocyte development. However,no significant variation was observed across all stages of testis development,although the expression level in the spermatogenesis stage was relatively low,which may result from the regulation of FST by aromatase.展开更多
基金supported by grants from the National Key R&D Program of China(2020YFA0509500)the National Natural Science Foundation of China(U19A2036,32102512,31872335,and 31802044)+1 种基金the National Special Foundation for Transgenic Species of China(2014ZX0800605B)the Sichuan Science and Technology Program,China(2021YFYZ0009 and 2021YFYZ0030)。
文摘Follistatin(FST) is an important regulator of skeletal muscle growth and adipose deposition through its ability to bind to several members of the transforming growth factor-β(TGF-β) superfamily, and thus may be a good candidate for future animal breeding programs. However, the molecular mechanisms underlying the phenotypic changes have yet to be clarified in pig. We generated transgenic(TG) pigs that express human FST specifically in skeletal muscle tissues and characterized the phenotypic changes compared with the same tissues in wild-type pigs. The TG pigs showed increased skeletal muscle growth, decreased adipose deposition, and improved metabolism status(P<0.05). Transcriptome analysis detected important roles of the PIK3–AKT signaling pathway, calcium-mediated signaling pathway, and amino acid metabolism pathway in FST-induced skeletal muscle hypertrophy, and depot-specific oxidative metabolism changes in psoas major muscle. Furthermore, the lipid metabolism-related process was changed in adipose tissue in the TG pigs. Gene set enrichment analysis revealed that genes related to lipid synthesis, lipid catabolism, and lipid storage were down-regulated(P<0.01) in the TG pigs for subcutaneous fat, whereas genes related to lipid catabolism were significantly up-regulated(P<0.05) in the TG pigs for retroperitoneal fat compared with their expression levels in wild-type pigs. In liver, genes related to the TGF-β signaling pathway were over-represented in the TG pigs, which is consistent with the inhibitory role of FST in regulating TGF-β signaling. Together, these results provide new insights into the molecular mechanisms underlying the phenotypic changes in pig.
基金supported by the National Natural Science Foundation of China (31301955)the China Agriculture Research System (CARS-36)
文摘Follistatin （FS） is a secreted protein, which was originally isolated from porcine follicular fluid. Expression of follistatin is tightly regulated during porcine growth and development. To study the essential transcriptional regions of the porcine FS promoter, ten primer pairs were designed to amplify segments with different lengths of the FS promoter from -1 800 to ＋16 bp. The products were then inserted into the pGL3-basic vector to analyze the relative luciferase activity. The results showed that the most remarkable changes of promoter activity were observed between constructs （-302/＋16 bp）-FS and （-180/＋16 bp）-FS （P〈0.01）. Further research showed that the reconstructed reporter plasmid lacking myeloid zinc finger 1 （MZF1） binding sequence had significantly decreased luciferase activity （P〈0.05）. Furthermore, the FS protein expression was significantly increased in PK15 cells while the MZF1 was overexpressed, suggesting that the short sequence ＂TCCCCACC＂ （the recognition site of transcription factor MZF1） was the most important for FS transcription activation in the porcine.
基金supported by the Project of Young and Middle-aged Scientific Research Fund of Wannan Medical College(No.WK2019F11).
文摘Objective:Follistatin(FST)inhibits the action of activin by interfering with the binding of activin to its receptor.Although the prognostic value of FST in various cancers has been investigated previously,studies rarely focused on hypopharyngeal carcinoma(HPC).In our study,the effect of FST expression on HPC tissues and cell lines was investigated.Methods:A total of 60 patients with HPC were recruited for this study.Levels of FST mRNA and protein were measured by quantitative polymerase chain reaction(PCR)and immunohistochemistry in HPC tissue samples and by qPCR in the HPC FaDu cells,as well as immortal nasopharyngeal epithelial cell line NP-69 cells.After silencing the FST expression in FaDu cells using lentivirus-mediated siRNA that was specific for FST mRNA,cell proliferation was determined by a Celigo assay.Tumor growth was monitored in nude mice and viability was determined by a methylthiazoletetrazolium assay.The ratio of cell cycle arrest and apoptosis was evaluated by flow cytometry.The colony formation ability was performed using Giemsa staining.In addition,wound healing and Transwell migration and invasion assays were performed for the analysis of cell motility.Results:FST expression was significantly higher in human HPC tissue and FaDu cells than in normal tissue and NP-69 cells.A higher expression of FST in HPC samples was positively correlated with advanced tumors.Moreover,FST knockdown by shRNA significantly decreased cell growth,colony formation,migration and invasion.Furthermore,FST silencing increased the cell apoptosis percentage,and arrested cell cycle in the S phase in FaDu cells.In addition,FST silencing suppressed tumor growth in vivo.Conclusions:Our results indicated that the FST gene was associated with HPC progression and may serve as a potential therapeutic target for the treatment of HPC.
文摘Objective: To investigate the effects of follistatin(rhFS-288) on biosynthesis andsecretion of testosterone in rat Leydig cell in vitro.Methods: Leydig cells were isolated from Wistar rat testes by a discontinuous Per-coll gradient procedure. Purified cells were incubated in 24-well plate(105 cell/ml/well)and maintained for 24 h in a CO2 incubator. rhFS-288 and Ca2+ were added to the wellsindependently or jointly in both baseline (without hCG) and stimulation condition (1.0IU/ml of hCG) to observe the change of testosterone concentration in the media.Results: rhFS-288 showed a dose-dependent inhibiting effect on testosterone releasein baseline and stimulating condition. Ca2+ presented inhibitory effect either. Whereas,escape phenomenon emerged while Ca2+ concentration reached to 100 mmol/L. A com-bination of rhFS-288 with Ca2+ displayed a dose-dependent inhibition on testosterone se-cretion.Conclusion: rhFS-288 inhibits testosterone secretion in a dose-dependent manner.Calcium is thought to be the second messenger of FS action. The mechanism of escapephenomenon during high dose of Ca2+ along is unknown.
文摘Molecular biomarkers are commonly used for the management of several types of malignant tumours in routine clinical practice. However, this is not the case for malignant gliomas. Cytokines and Angiogenesis factors are potential candidates due to their intrinsic role in tumourigenesis. Pre- and post-operative serum from 36 malignant glioma patients and 36 controls was analysed using the Bio-Plex Pro Angiogenesis and Cytokines Assay (Bio-Rad, USA). Amongst the molecules tested, the serum concentration of follistatin was significantly higher in patients than in controls. Moreover, the serum concentration of follistatin of the patients postoperatively was significantly reduced compared to that preoperatively. Factors such as age and gender did not affect the concentrations of follistatin measured in the serum of patients pre- and post-operatively as well as healthy controls. This is the first report of follistatin as potential biomarker for the detection of malignant gliomas.
基金Supported by the National High Technology Research and Development Program of China(863 Program)(No.2012AA10A403)
文摘Follistatin(FST) is a monomeric glycoprotein highly enriched in cysteines and belongs to TGF-β superfamily. FST can suppress the secretion of follicle-stimulating hormone and plays a vital role in the reproduction of vertebrates. We used rapid amplification of cDNA ends technology to clone the FST gene of half-smooth tongue sole,Cynoglossus semilaevis. We characterized its phylogenetic context and expression patterns to elucidate its function in the breeding season. The full-length sequence of FST is 1 455 bp and encodes a protein of 321 amino acids. We investigated the expression pattern of FST in C. semilaevis at different stages of reproduction using reverse transcription-polymerase chain reaction(RTPCR). FST m RNA was expressed in all 13 tissues analyzed,and was expressed at high levels in gonad and at slightly lower levels in gill and brain. During the reproductive cycle of C. semilaevis,the transcript level of FST was the highest in the perinucleolus stage,decreased in the primary yolk stage,slightly increased in the tertiary yolk stage,and then reduced to a minimal level in the atretic follicles stage of the ovary. We concluded that FST suppressed follicle-stimulating hormone,which stimulated oocyte development. However,no significant variation was observed across all stages of testis development,although the expression level in the spermatogenesis stage was relatively low,which may result from the regulation of FST by aromatase.
