Mitogen-activated protein kinase-8-interacting protein 2(MAPK8IP2)is a scaffold protein that modulates MAPK signal cascades.Although MAPK pathways were heavily implicated in prostate cancer progression,the regulation ...Mitogen-activated protein kinase-8-interacting protein 2(MAPK8IP2)is a scaffold protein that modulates MAPK signal cascades.Although MAPK pathways were heavily implicated in prostate cancer progression,the regulation of MAPK8IP2 expression in prostate cancer is not yet reported.We assessed MAPK8IP2 gene expression in prostate cancer related to disease progression and patient survival outcomes.MAPK8IP2 expression was analyzed using multiple genome-wide gene expression datasets derived from The Cancer Genome Atlas(TCGA)RNA-sequence project and complementary DNA(cDNA)microarrays.Multivariable Cox regressions and log-rank tests were used to analyze the overall survival outcome and progression-free interval.MAPK8IP2 protein expression was evaluated using the immunohistochemistry approach.The quantitative PCR and Western blot methods analyzed androgen-stimulated MAPK8IP2 expression in LNCaP cells.In primary prostate cancer tissues,MAPK8IP2 mRNA expression levels were significantly higher than those in the case-matched benign prostatic tissues.Increased MAPK8IP2 expression was strongly correlated with late tumor stages,lymph node invasion,residual tumors after surgery,higher Gleason scores,and preoperational serum prostate-specific antigen(PSA)levels.MAPK8IP2 upregulation was significantly associated with worse overall survival outcomes and progression-free intervals.In castration-resistant prostate cancers,MAPK8IP2 expression strongly correlated with androgen receptor(AR)signaling activity.In cell culture-based experiments,MAPK8IP2 expression was stimulated by androgens in AR-positive prostate cancer cells.However,MAPK8IP2 expression was blocked by AR antagonists only in androgen-sensitive LNCaP but not castration-resistant C4-2B and 22RV1 cells.These results indicate that MAPK8IP2 is a robust prognostic factor and therapeutic biomarker for prostate cancer.The potential role of MAPK8IP2 in the castration-resistant progression is under further investigation.展开更多
The VirE2-interaction protein 1(VIP1)serves as a regulator of mitogen-activated protein kinase 3(MPK3)-mediated stress gene modulation under biotic stress,which in turn activates the MPK3 pathway in Arabidopsis.The mo...The VirE2-interaction protein 1(VIP1)serves as a regulator of mitogen-activated protein kinase 3(MPK3)-mediated stress gene modulation under biotic stress,which in turn activates the MPK3 pathway in Arabidopsis.The mode of action of the VIP1 protein in Populus in response to biotic stress remains unknown.In this study,we cloned the full-length cDNA of the PtVIP1 gene from Populus trichocarpa(accession number of GenBank:KY793105).The VIP1 protein harboured a conserved bZIP(basic leucine zipper)domain located in the C-terminus.The VIP1 subcellular localization assay indicated that the VIP1 protein was present in the cytoplasm and nucleus under normal conditions,and that an increase in the amount of the protein in the nucleus occurred after treatment with flg22,the elicitor-active epitope of flagellin which triggers the innate immune response in plants.Transgenic Populus plants overexpressing VIP1 genes(PtVIP1 of Populus;or AtVIP1 of Arabidopsis,as positive control)were generated to investigate the role of VIP1 in vivo.The expression of poplar pathogenesis-related protein 1(PR1)genes was upregulated in transgenic-PtVIP1 or AtVIP1 poplar plants.The transgenic poplar plants overexpressing PtVIP1 or AtVIP1 also showed enhanced resistance to Brenneria salicis infection.These results suggest that the VIP1 protein accumulates in the nucleus in response to biotic stress,and that the pathogen resistance of transgenic VIP1 poplar may be associated with the induced expression of PR1 genes in response to pathogen challenge.展开更多
Background:The casein kinase 2-interacting protein-1(CKIP-1)is important in the development of osteoblasts and cardiomyocytes.However,the effects of CKIP-1 on osteoblast precursor mesenchymal stem cells(MSCs)remain un...Background:The casein kinase 2-interacting protein-1(CKIP-1)is important in the development of osteoblasts and cardiomyocytes.However,the effects of CKIP-1 on osteoblast precursor mesenchymal stem cells(MSCs)remain unclear.This study aimed to determine whether CKIP-1 affects osteogenic differentiation in MSCs and explore the relationship of CKIP-1 and inflammation.Methods:Bone marrow MSCs of CKIP-1 wild type(WT)and knockout(KO)mice were cultivated in vitro.Cell phenotype was analyzed by flow cytometry,colony formation was detected to study the proliferative ability.Osteogenic and adipogenic induction were performed.The osteogenic ability was explored by alizarin red staining,alkaline phosphatase(ALP)staining and ALP activity detection.Quantitative real-time polymerase chain reaction(qRT-PCR)was carried out to determine the mRNA expression levels of osteoblast marker genes.The adipogenic ability was detected by oil red O staining.Content of the bone was analyzed to observe the differences of bone imaging parameters including trabecular bone volume/tissue volume(BV/TV),bone surface area fraction/trabecular BV,trabecular number(Tb.N),and trabecular spacing(Tb.sp).Interleukin(IL)-1b was injected on WT mice of 2 months old and 18 months old,respectively.Difference in CKIP-1 expression was detected by RT-PCR and western blot.The relationship between CKIP-1 and inflammation was explored by RT-PCR and western blot.Results:ALP assays,alizarin red staining,and qRT-PCR showed that MSCs derived from CKIP-1 KO mice exhibited a stronger capability for osteogenesis.Micro-computed tomography detection showed that among 18-month-old mice,CKIP-1 KO mice presented significantly higher bone mass compared withWTmice(P=0.02).No significant difference was observed in 2-month-old mice.In vivo data showed that expression of CKIP-1 was higher in the bone marrow of aging mice than in young mice(4.3-fold increase at themRNA level,P=0.04).Finally,the expression levels of CKIP-1 in bone marrow(3.2-fold increase at themRNA level,P=0.03)and cultured MSCs were up-regulated on chronic inflammatory stimulation by IL-1b.Conclusions:CKIP-1 is responsible for negative regulation of MSC osteogenesis with age-dependent effects.Increasing levels of inflammation with aging may be the primary factor responsible for higher expression levels of CKIP-1 but may not necessarily affect MSC aging.展开更多
Objective Colorectal cancer(CRC),a prevalent malignancy worldwide,has prompted extensive research into anticancer drugs.Traditional Chinese medicinal materials offer promising avenues for cancer management due to thei...Objective Colorectal cancer(CRC),a prevalent malignancy worldwide,has prompted extensive research into anticancer drugs.Traditional Chinese medicinal materials offer promising avenues for cancer management due to their diverse pharmacological activities.This study investigated the effects of Notopterygium incisum,a traditional Chinese medicine named Qianghuo(QH),on CRC cells and the underlying mechanism.Methods The sulforhodamine B assay and colony formation assay were employed to assess the effect of QH extract on the proliferation of CRC cell lines HCT116 and Caco-2.Propidium iodide(PI)staining was utilized to detect cell cycle progression,and PE Annexin V staining to detect apoptosis.Western blotting was conducted to examine the levels of apoptotic proteins,including B-cell lymphoma 2-interacting mediator of cell death(BIM),B-cell lymphoma 2(Bcl-2),Bcl-2-associated X protein(BAX)and cleaved caspase-3,as well as BIM stability after treatment with the protein synthesis inhibitor cycloheximide.The expression of BAX was suppressed using lentivirus-mediated shRNA to validate the involvement of the BIM/BAX axis in QH-induced apoptosis.The in vivo effects of QH extract on tumor growth were observed using a xenograft model.Lastly,APC^(Min+)mice were used to study the effects of QH extract on primary intestinal tumors.Results QH extract exhibited significant in vitro anti-CRC activities evidenced by the inhibition of cell proliferation,perturbation of cell cycle progression,and induction of apoptosis.Mechanistically,QH extract significantly increased the stability of BIM proteins,which undergo rapid degradation under unstressed conditions.Knockdown of BAX,the downstream effector of BIM,significantly rescued QH-induced apoptosis.Furthermore,the in vitro effect of QH extract was recapitulated in vivo.QH extract significantly inhibited the tumor growth of HCT116 xenografts in nude mice and decreased the number of intestinal polyps in the APC^(Min+)mice.Conclusion QH extract promotes the apoptosis of CRC cells by preventing the degradation of BIM.展开更多
文摘Mitogen-activated protein kinase-8-interacting protein 2(MAPK8IP2)is a scaffold protein that modulates MAPK signal cascades.Although MAPK pathways were heavily implicated in prostate cancer progression,the regulation of MAPK8IP2 expression in prostate cancer is not yet reported.We assessed MAPK8IP2 gene expression in prostate cancer related to disease progression and patient survival outcomes.MAPK8IP2 expression was analyzed using multiple genome-wide gene expression datasets derived from The Cancer Genome Atlas(TCGA)RNA-sequence project and complementary DNA(cDNA)microarrays.Multivariable Cox regressions and log-rank tests were used to analyze the overall survival outcome and progression-free interval.MAPK8IP2 protein expression was evaluated using the immunohistochemistry approach.The quantitative PCR and Western blot methods analyzed androgen-stimulated MAPK8IP2 expression in LNCaP cells.In primary prostate cancer tissues,MAPK8IP2 mRNA expression levels were significantly higher than those in the case-matched benign prostatic tissues.Increased MAPK8IP2 expression was strongly correlated with late tumor stages,lymph node invasion,residual tumors after surgery,higher Gleason scores,and preoperational serum prostate-specific antigen(PSA)levels.MAPK8IP2 upregulation was significantly associated with worse overall survival outcomes and progression-free intervals.In castration-resistant prostate cancers,MAPK8IP2 expression strongly correlated with androgen receptor(AR)signaling activity.In cell culture-based experiments,MAPK8IP2 expression was stimulated by androgens in AR-positive prostate cancer cells.However,MAPK8IP2 expression was blocked by AR antagonists only in androgen-sensitive LNCaP but not castration-resistant C4-2B and 22RV1 cells.These results indicate that MAPK8IP2 is a robust prognostic factor and therapeutic biomarker for prostate cancer.The potential role of MAPK8IP2 in the castration-resistant progression is under further investigation.
基金supported by the National Natural Science Foundation of China(Grant No.31570639)the Distinguished Young Scholars Fund of Nanjing Forestry University,the Priority Academic Program Development of Nanjing Forestry University the Poplar Germplasm Nursery Project(Jiangsu Provincial Platform for Conservation and Utilizations for Agricultural Germplasm)
文摘The VirE2-interaction protein 1(VIP1)serves as a regulator of mitogen-activated protein kinase 3(MPK3)-mediated stress gene modulation under biotic stress,which in turn activates the MPK3 pathway in Arabidopsis.The mode of action of the VIP1 protein in Populus in response to biotic stress remains unknown.In this study,we cloned the full-length cDNA of the PtVIP1 gene from Populus trichocarpa(accession number of GenBank:KY793105).The VIP1 protein harboured a conserved bZIP(basic leucine zipper)domain located in the C-terminus.The VIP1 subcellular localization assay indicated that the VIP1 protein was present in the cytoplasm and nucleus under normal conditions,and that an increase in the amount of the protein in the nucleus occurred after treatment with flg22,the elicitor-active epitope of flagellin which triggers the innate immune response in plants.Transgenic Populus plants overexpressing VIP1 genes(PtVIP1 of Populus;or AtVIP1 of Arabidopsis,as positive control)were generated to investigate the role of VIP1 in vivo.The expression of poplar pathogenesis-related protein 1(PR1)genes was upregulated in transgenic-PtVIP1 or AtVIP1 poplar plants.The transgenic poplar plants overexpressing PtVIP1 or AtVIP1 also showed enhanced resistance to Brenneria salicis infection.These results suggest that the VIP1 protein accumulates in the nucleus in response to biotic stress,and that the pathogen resistance of transgenic VIP1 poplar may be associated with the induced expression of PR1 genes in response to pathogen challenge.
基金Supported by the grants from the National Key Research and Development Project(No.2017YFB1304300)Conversion Fund of PLA General Hospital(No.2017tm-018)the Clinical Research Support Fund of PLA General Hospital(No.2017fc-tsys-2013).
文摘Background:The casein kinase 2-interacting protein-1(CKIP-1)is important in the development of osteoblasts and cardiomyocytes.However,the effects of CKIP-1 on osteoblast precursor mesenchymal stem cells(MSCs)remain unclear.This study aimed to determine whether CKIP-1 affects osteogenic differentiation in MSCs and explore the relationship of CKIP-1 and inflammation.Methods:Bone marrow MSCs of CKIP-1 wild type(WT)and knockout(KO)mice were cultivated in vitro.Cell phenotype was analyzed by flow cytometry,colony formation was detected to study the proliferative ability.Osteogenic and adipogenic induction were performed.The osteogenic ability was explored by alizarin red staining,alkaline phosphatase(ALP)staining and ALP activity detection.Quantitative real-time polymerase chain reaction(qRT-PCR)was carried out to determine the mRNA expression levels of osteoblast marker genes.The adipogenic ability was detected by oil red O staining.Content of the bone was analyzed to observe the differences of bone imaging parameters including trabecular bone volume/tissue volume(BV/TV),bone surface area fraction/trabecular BV,trabecular number(Tb.N),and trabecular spacing(Tb.sp).Interleukin(IL)-1b was injected on WT mice of 2 months old and 18 months old,respectively.Difference in CKIP-1 expression was detected by RT-PCR and western blot.The relationship between CKIP-1 and inflammation was explored by RT-PCR and western blot.Results:ALP assays,alizarin red staining,and qRT-PCR showed that MSCs derived from CKIP-1 KO mice exhibited a stronger capability for osteogenesis.Micro-computed tomography detection showed that among 18-month-old mice,CKIP-1 KO mice presented significantly higher bone mass compared withWTmice(P=0.02).No significant difference was observed in 2-month-old mice.In vivo data showed that expression of CKIP-1 was higher in the bone marrow of aging mice than in young mice(4.3-fold increase at themRNA level,P=0.04).Finally,the expression levels of CKIP-1 in bone marrow(3.2-fold increase at themRNA level,P=0.03)and cultured MSCs were up-regulated on chronic inflammatory stimulation by IL-1b.Conclusions:CKIP-1 is responsible for negative regulation of MSC osteogenesis with age-dependent effects.Increasing levels of inflammation with aging may be the primary factor responsible for higher expression levels of CKIP-1 but may not necessarily affect MSC aging.
基金funded by the Science and Technology Development Fund,Macao SAR(No.0105/2022/A2 and No.006/2023/SKL).
文摘Objective Colorectal cancer(CRC),a prevalent malignancy worldwide,has prompted extensive research into anticancer drugs.Traditional Chinese medicinal materials offer promising avenues for cancer management due to their diverse pharmacological activities.This study investigated the effects of Notopterygium incisum,a traditional Chinese medicine named Qianghuo(QH),on CRC cells and the underlying mechanism.Methods The sulforhodamine B assay and colony formation assay were employed to assess the effect of QH extract on the proliferation of CRC cell lines HCT116 and Caco-2.Propidium iodide(PI)staining was utilized to detect cell cycle progression,and PE Annexin V staining to detect apoptosis.Western blotting was conducted to examine the levels of apoptotic proteins,including B-cell lymphoma 2-interacting mediator of cell death(BIM),B-cell lymphoma 2(Bcl-2),Bcl-2-associated X protein(BAX)and cleaved caspase-3,as well as BIM stability after treatment with the protein synthesis inhibitor cycloheximide.The expression of BAX was suppressed using lentivirus-mediated shRNA to validate the involvement of the BIM/BAX axis in QH-induced apoptosis.The in vivo effects of QH extract on tumor growth were observed using a xenograft model.Lastly,APC^(Min+)mice were used to study the effects of QH extract on primary intestinal tumors.Results QH extract exhibited significant in vitro anti-CRC activities evidenced by the inhibition of cell proliferation,perturbation of cell cycle progression,and induction of apoptosis.Mechanistically,QH extract significantly increased the stability of BIM proteins,which undergo rapid degradation under unstressed conditions.Knockdown of BAX,the downstream effector of BIM,significantly rescued QH-induced apoptosis.Furthermore,the in vitro effect of QH extract was recapitulated in vivo.QH extract significantly inhibited the tumor growth of HCT116 xenografts in nude mice and decreased the number of intestinal polyps in the APC^(Min+)mice.Conclusion QH extract promotes the apoptosis of CRC cells by preventing the degradation of BIM.
