We previously demonstrated that matrine could inhibit the proliferating, migrating, as well as invading processes of both PC-3 and DU145 cells. However, the underlying molecular mechanisms have not yet been clearly de...We previously demonstrated that matrine could inhibit the proliferating, migrating, as well as invading processes of both PC-3 and DU145 cells. However, the underlying molecular mechanisms have not yet been clearly defined. In this study, using various techniques such as high throughput sequencing technology, bioinformatics, quantitative real-time PCR, and immunoblot analysis,we aimed to understand whether matrine serves as a novel regulator of FOXO and PI3K-AKT signaling pathway. DU145 and PC-3 cell lines were cultured for 24 h in vitro. Cells were treated with either matrine or control serum for 48 h, followed by extraction of total RNA. The RNA was sequenced using HiSeq 2500 high-throughput sequencing platform (Illumina). A gene library was established and quality analysis of read data carried out. Integrated database from the website DAVID was used to analyze Gene Ontology (GO), and Kyoto encyclopedia of genes and genomes (KEGG) pathway of differential genes was used for pathway analysis, screening for fold differences of more than two times. The FOXO and PI3K-AKT signaling pathways were screened, and expression levels of mRNA and core protein detected by real-time PCR and immunoblotting, respectively. High throughput sequencing and GO analysis revealed that differentially expressed genes before and after treatment played an important role in cell metabolic process, growth process, anatomical structure formation, cellular component organization, and biological regulation. KEGG signal pathway analysis revealed that FOXO and PI3K-AKT signal pathways had a significant difference between before and after matrine-treated androgen-independent prostate cancer cells PC-3 and DU145. Real-time PCR showed that matrine treatment led to a significant increase in the expression levels of FOXO1A, FOXO3A, FOXO4, and FOXO6 in DU145 and PC-3 cells (P<0.01 or P<0.05), whereas the PI3K expression levels decreased (P<0.01). Similarly, immunoblotting revealed a significant increase (P<0.05) in the expression levels of FOXO1A FOXO3展开更多
Prostate cancer(PCa)is the most common malignancy in men.Despite aggressive therapy involving surgery and hormonal treatments,the recurrence and emergence of metastatic castration-resistant prostate cancer(CRPCa)remai...Prostate cancer(PCa)is the most common malignancy in men.Despite aggressive therapy involving surgery and hormonal treatments,the recurrence and emergence of metastatic castration-resistant prostate cancer(CRPCa)remain a major challenge.Dysregulation of the transforming growth factor-β(TGF-β)signaling pathway is crucial to PCa development and progression.This also contributes to androgen receptor activation and the emergence of CRPC.In addition,TGF-β signaling regulates long non-coding RNA(lncRNA)expression in multiple cancers,including PCa.Here,we discuss the complex regulatory network of lncRNAs and TGF-β signaling in PCa and their potential applications in diagnosing,prognosis,and treating PCa.Further investigations on the role of lncRNAs in the TGF-β pathway will help to better understand PCa pathogenesis.展开更多
We sought to investigate the underlying mechanism of action of the long noncoding RNA （IncRNA） LOC283070 in the development of androgen independence in prostate cancer. The interactions between LOC283070 and target ...We sought to investigate the underlying mechanism of action of the long noncoding RNA （IncRNA） LOC283070 in the development of androgen independence in prostate cancer. The interactions between LOC283070 and target proteins were investigated by RNA pull-down and RNA-binding protein immunoprecipitation （RIP） assays. Subceilular fractionation and quantitative reverse transcription-polymerase chain reaction （qRT-PCR） were used to detect the subcellular localization of LOC283070. Western blotting was performed to detect the expression of prohibitin 2 （PHB2）. Luciferase activity assays were performed to evaluate the effects of LOC283070 and PHB2 on the androgen receptor （AR） signaling pathway. A methyl thiazolyl tetrazolium （MTT） assay and a growth curve assay were used to test cell viability. Flow cytometry was performed to analyze cell cycles. A transwell assay was employed to test cell migration. We identified PHB2 as an interaction partner of LOC283070 in the pull-down and RIP experiments. Furthermore, we confirmed that the enrichment of L0C283070 with PHB2 in androgen-independent LNCaP （LNCaP-AI） cells was much greater than that in LNCaP cells. Moreover, the expression of PHB2 was not significantly different between the two cell lines, and the expression of LOC283070 in the nuclei of the LNCaP-AI cells was significantly greater than that in the LNCaP cells. In vitro data revealed that PHB2 overexpression significantly inhibited AR activity and cell proliferation and migration and induced accumulation of prostate cancer cells in GO/G1 phase. Moreover, the overexpression of LOC283070 fully abrogated the effects of PHB2 overexpression. In conclusion, we found that LOC283070 can bind to PHB2 located in the nucleus and inhibit its effect, and this is one of the mechanisms by which LOC283070 is involved in the transition of LNCaP cells into androgen-independent cells.展开更多
The anti-tumor activity of curcumin against androgen-independent prostate cancer cells in vitro and the possible mechanism were investigated.After curcumin treatment,the effect of curcumin on the proliferation of pros...The anti-tumor activity of curcumin against androgen-independent prostate cancer cells in vitro and the possible mechanism were investigated.After curcumin treatment,the effect of curcumin on the proliferation of prostate cancer PC-3 cells was assessed by CFSE staining.Flow cytometery（FCM） was performed to analyze the cell cycle and the induction of apoptosis of tumor cells.A luciferase reporter gene assay was used to determine the effects of curcumin on the activities of intracellular NF-κB and AP-1 signaling pathways.The results showed curcumin could effectively inhibit the proliferation of PC-3 cells in vitro（P0.05）.Cells were arrested at G2/M phase.After curcumin treatment,the percentage of apoptotic cells was significantly higher than in control group（P0.05）.The results of the luciferase assay revealed that curcumin selectively inhibited the activities of the NF-κB and AP-1 signaling pathways in PC-3 cells significantly.It was suggested that curcumin could exert anti-tumor activity against androgen-independent prostate cancer cells in vitro by inhibiting cellular proliferation and inducing apoptosis,which was probably contributed to the inhibition of transcription factors NF-κB and AP-1.展开更多
Objective: To identify mutually regulated proteins in PC-3 and DU145 androgen-independent prostate cancer cell lines treated with 1,5-bis(2-hydroxyphenyl)-1,4-pentadiene-3-one(MS17), and to study the molecular pathway...Objective: To identify mutually regulated proteins in PC-3 and DU145 androgen-independent prostate cancer cell lines treated with 1,5-bis(2-hydroxyphenyl)-1,4-pentadiene-3-one(MS17), and to study the molecular pathways that contributed to the anticancer activity of MS17.Methods: PC-3 and DU145 cells were treated with 3 × EC_(50)(15 μM) concentration of MS17 for 24 h and were subjected to protein expression profiling using two-dimensional gel electrophoresis and protein identification by mass spectrometry.Selected differentially expressed proteins with significant P-value of P<0.05 and fold change over 1.5-folds were filtered through and ontologically classified.Mutually regulated proteins were ranked by fold change and identified as common protein targets of MS17.Results: Profiling data revealed that, the mutually down-regulated proteins included ACTB and ACTG associated with structural molecule activity, ACTN1 with cell cycle, ACTN4 with cell migration, HNRPK with apoptosis, PLST with morphogenesis and TERA with proteolysis.However, the expressions of CH60 and HS71 A respectively associated with response to unfolded protein demonstrated opposing regulation in PC-3 and DU145 cells.Pathway analysis of the differentially expressed proteins in PC-3 cells demonstrated the modulation of top pathways associated with cell-cell adhesion and cytoskeletal organization while in DU145 cells the pathways were associated with proteosomal degradation, regulation of electrolytes and water, regulation control of germ cells and organization of filament assembly/disassembly.Conclusions: The findings of the present study provide an understanding on the anti-tumorigenic activity of MS17 at the proteome level and warrant further research for its potential application for the management and treatment of androgen-independent prostate cancer.展开更多
Prostate cancer(PC) is a common malignant tumor in the male seniors,and its incidence has been ascending in China in recent years. Over 80%of the patients are diagnosed as being in the late stage and lose the opportun...Prostate cancer(PC) is a common malignant tumor in the male seniors,and its incidence has been ascending in China in recent years. Over 80%of the patients are diagnosed as being in the late stage and lose the opportunity of radical operation because of its early stage metastasis. Endocrinotherapy is regarded展开更多
基金supported by the National Natural Science Foundation of China (81472382)the National Natural Science Foundation of China for Young Scientists (81101947)+3 种基金the Guangdong Province Natural Science Foundation (2014A030313079)the Fundamental Research Funds for the Central Universities (14ykpy19)Guangdong Province Science and Technology for Social Development Project (2013B021800107)Guangzhou City in 2015 scientific research projects (7415600066401 to Hai Huang)
文摘We previously demonstrated that matrine could inhibit the proliferating, migrating, as well as invading processes of both PC-3 and DU145 cells. However, the underlying molecular mechanisms have not yet been clearly defined. In this study, using various techniques such as high throughput sequencing technology, bioinformatics, quantitative real-time PCR, and immunoblot analysis,we aimed to understand whether matrine serves as a novel regulator of FOXO and PI3K-AKT signaling pathway. DU145 and PC-3 cell lines were cultured for 24 h in vitro. Cells were treated with either matrine or control serum for 48 h, followed by extraction of total RNA. The RNA was sequenced using HiSeq 2500 high-throughput sequencing platform (Illumina). A gene library was established and quality analysis of read data carried out. Integrated database from the website DAVID was used to analyze Gene Ontology (GO), and Kyoto encyclopedia of genes and genomes (KEGG) pathway of differential genes was used for pathway analysis, screening for fold differences of more than two times. The FOXO and PI3K-AKT signaling pathways were screened, and expression levels of mRNA and core protein detected by real-time PCR and immunoblotting, respectively. High throughput sequencing and GO analysis revealed that differentially expressed genes before and after treatment played an important role in cell metabolic process, growth process, anatomical structure formation, cellular component organization, and biological regulation. KEGG signal pathway analysis revealed that FOXO and PI3K-AKT signal pathways had a significant difference between before and after matrine-treated androgen-independent prostate cancer cells PC-3 and DU145. Real-time PCR showed that matrine treatment led to a significant increase in the expression levels of FOXO1A, FOXO3A, FOXO4, and FOXO6 in DU145 and PC-3 cells (P<0.01 or P<0.05), whereas the PI3K expression levels decreased (P<0.01). Similarly, immunoblotting revealed a significant increase (P<0.05) in the expression levels of FOXO1A FOXO3
基金supported by intramural funds from BITS Pilani,Hyderabad,India.
文摘Prostate cancer(PCa)is the most common malignancy in men.Despite aggressive therapy involving surgery and hormonal treatments,the recurrence and emergence of metastatic castration-resistant prostate cancer(CRPCa)remain a major challenge.Dysregulation of the transforming growth factor-β(TGF-β)signaling pathway is crucial to PCa development and progression.This also contributes to androgen receptor activation and the emergence of CRPC.In addition,TGF-β signaling regulates long non-coding RNA(lncRNA)expression in multiple cancers,including PCa.Here,we discuss the complex regulatory network of lncRNAs and TGF-β signaling in PCa and their potential applications in diagnosing,prognosis,and treating PCa.Further investigations on the role of lncRNAs in the TGF-β pathway will help to better understand PCa pathogenesis.
基金This work was supported by grants from the National Natural Science Foundation of China （No. 81702538, 81372764, and 81528015） and the Natural Science Foundation of Shandong Province （No. ZR201702160271, ZR2018MH030, and ZR2018MH025）.
文摘We sought to investigate the underlying mechanism of action of the long noncoding RNA （IncRNA） LOC283070 in the development of androgen independence in prostate cancer. The interactions between LOC283070 and target proteins were investigated by RNA pull-down and RNA-binding protein immunoprecipitation （RIP） assays. Subceilular fractionation and quantitative reverse transcription-polymerase chain reaction （qRT-PCR） were used to detect the subcellular localization of LOC283070. Western blotting was performed to detect the expression of prohibitin 2 （PHB2）. Luciferase activity assays were performed to evaluate the effects of LOC283070 and PHB2 on the androgen receptor （AR） signaling pathway. A methyl thiazolyl tetrazolium （MTT） assay and a growth curve assay were used to test cell viability. Flow cytometry was performed to analyze cell cycles. A transwell assay was employed to test cell migration. We identified PHB2 as an interaction partner of LOC283070 in the pull-down and RIP experiments. Furthermore, we confirmed that the enrichment of L0C283070 with PHB2 in androgen-independent LNCaP （LNCaP-AI） cells was much greater than that in LNCaP cells. Moreover, the expression of PHB2 was not significantly different between the two cell lines, and the expression of LOC283070 in the nuclei of the LNCaP-AI cells was significantly greater than that in the LNCaP cells. In vitro data revealed that PHB2 overexpression significantly inhibited AR activity and cell proliferation and migration and induced accumulation of prostate cancer cells in GO/G1 phase. Moreover, the overexpression of LOC283070 fully abrogated the effects of PHB2 overexpression. In conclusion, we found that LOC283070 can bind to PHB2 located in the nucleus and inhibit its effect, and this is one of the mechanisms by which LOC283070 is involved in the transition of LNCaP cells into androgen-independent cells.
基金supported by a grant from the Specialized Research Fund for the Doctoral Program of Higher Education of China (No. 20090142120096)
文摘The anti-tumor activity of curcumin against androgen-independent prostate cancer cells in vitro and the possible mechanism were investigated.After curcumin treatment,the effect of curcumin on the proliferation of prostate cancer PC-3 cells was assessed by CFSE staining.Flow cytometery（FCM） was performed to analyze the cell cycle and the induction of apoptosis of tumor cells.A luciferase reporter gene assay was used to determine the effects of curcumin on the activities of intracellular NF-κB and AP-1 signaling pathways.The results showed curcumin could effectively inhibit the proliferation of PC-3 cells in vitro（P0.05）.Cells were arrested at G2/M phase.After curcumin treatment,the percentage of apoptotic cells was significantly higher than in control group（P0.05）.The results of the luciferase assay revealed that curcumin selectively inhibited the activities of the NF-κB and AP-1 signaling pathways in PC-3 cells significantly.It was suggested that curcumin could exert anti-tumor activity against androgen-independent prostate cancer cells in vitro by inhibiting cellular proliferation and inducing apoptosis,which was probably contributed to the inhibition of transcription factors NF-κB and AP-1.
基金financially supported by the Fundamental Research Grant Scheme,(FRGS/1/2016/SKK08/MUSM/02/1)under the Ministry of Higher Education,Malaysia
文摘Objective: To identify mutually regulated proteins in PC-3 and DU145 androgen-independent prostate cancer cell lines treated with 1,5-bis(2-hydroxyphenyl)-1,4-pentadiene-3-one(MS17), and to study the molecular pathways that contributed to the anticancer activity of MS17.Methods: PC-3 and DU145 cells were treated with 3 × EC_(50)(15 μM) concentration of MS17 for 24 h and were subjected to protein expression profiling using two-dimensional gel electrophoresis and protein identification by mass spectrometry.Selected differentially expressed proteins with significant P-value of P<0.05 and fold change over 1.5-folds were filtered through and ontologically classified.Mutually regulated proteins were ranked by fold change and identified as common protein targets of MS17.Results: Profiling data revealed that, the mutually down-regulated proteins included ACTB and ACTG associated with structural molecule activity, ACTN1 with cell cycle, ACTN4 with cell migration, HNRPK with apoptosis, PLST with morphogenesis and TERA with proteolysis.However, the expressions of CH60 and HS71 A respectively associated with response to unfolded protein demonstrated opposing regulation in PC-3 and DU145 cells.Pathway analysis of the differentially expressed proteins in PC-3 cells demonstrated the modulation of top pathways associated with cell-cell adhesion and cytoskeletal organization while in DU145 cells the pathways were associated with proteosomal degradation, regulation of electrolytes and water, regulation control of germ cells and organization of filament assembly/disassembly.Conclusions: The findings of the present study provide an understanding on the anti-tumorigenic activity of MS17 at the proteome level and warrant further research for its potential application for the management and treatment of androgen-independent prostate cancer.
文摘Prostate cancer(PC) is a common malignant tumor in the male seniors,and its incidence has been ascending in China in recent years. Over 80%of the patients are diagnosed as being in the late stage and lose the opportunity of radical operation because of its early stage metastasis. Endocrinotherapy is regarded
