We conducted a study concerning the suppressive mechanism of KAI1/CD82 on hepatoma cell metastasis.Hepatocyte growth factor(HGF)induces the migration of hepatoma cells through activation of cellular sphingosine kinase...We conducted a study concerning the suppressive mechanism of KAI1/CD82 on hepatoma cell metastasis.Hepatocyte growth factor(HGF)induces the migration of hepatoma cells through activation of cellular sphingosine kinase 1(SphK1).Adenovirus-mediated gene transfer of KAI1(Ad-KAI1)downregulates the SphK1 expression and suppresses the HGF-induced migration of SMMC-7721 human hepatocellcular carcinoma cells.Overexpression of KAI1/CD82 significantly elevates Sprouty2 at the protein level.Ablation of Sprouty2 with RNA interference can block the KAI1/CD82-induced suppression of hepatoma cell migration and downregulation of SphK1 expression.It is demonstrated that KAI1/CD82 suppresses HGF-induced migration of hepatoma cells via upregulation of Sprouty2.展开更多
BACKGROUND:In vitro experiments and mice models have confirmed the importance of Sprouty 2(Spry2) in inhibiting tumorigenesis and the progression of human cancer.However,the prognostic value of Spry2 in cancer patient...BACKGROUND:In vitro experiments and mice models have confirmed the importance of Sprouty 2(Spry2) in inhibiting tumorigenesis and the progression of human cancer.However,the prognostic value of Spry2 in cancer patients remains unknown.This study is aimed to investigate the clinical relevance and prognostic significance of Spry2 expression in patients with hepatocellular carcinoma(HCC).METHODS:With samples from 240 randomly-selected HCC patients who underwent surgery,immunohistochemistry was used to investigate Spry2 expression on tissue microarrays.The correlation of Spry2 expression with survival was estimated by the Kaplan-Meier method and univariate/multivariate Cox proportional hazard regression analysis.Spry2,ERK and phospho-ERK expression in HCC cell lines was detected by Western blotting.RESULTS:Among the patients,86.3%(207 of 240) exhibited down-regulation of Spry2 expression.Patients negative for Spry2 showed poorer survival(P=0.002) and increased recurrence(P=0.003).Multivariate analysis further established Spry2 as an independent predictor of postoperative recurrence in HCC patients(HR=1.47;95% CI,1.02-2.08;P=0.037).Downregulation of Spry2 was associated with highly malignant phenotypes like vascular invasion and advanced tumor stages,and was positively correlated with the metastatic potential of HCC cell lines.CONCLUSION:In the era of molecular targeted therapy,the expression of Spry2 in HCC may have relevant clinical significance and turn out to be a key factor in prognostic assessment and in treatment planning.展开更多
Objective:To investigate the effect of mitogen-activated protein kinase interaction serine kinase 1(Mnk1)gene deletion on lipopolysaccharide(LPS)-induced inflammatory response in mouse macrophages(Mφ)and the possible...Objective:To investigate the effect of mitogen-activated protein kinase interaction serine kinase 1(Mnk1)gene deletion on lipopolysaccharide(LPS)-induced inflammatory response in mouse macrophages(Mφ)and the possible mechanism.Methods:Healthy male wildtype C57BL/6J(WT)and Mnk1 knockout(KO)mice were selected at 8-10 weeks of age and divided into WT+PBS,KO+PBS,WT+LPS and KO+LPS groups,and the serum levels of IL-1βwere measured by ELISA after 24 h intraperitoneal injection of PBS or LPS.The mRNA expression levels of IL-1βand Sprouty2(Spry2)in the spleen Mφwere measured by qRTPCR.Mφwas also extracted from the peritoneal cavity of two strains of mice for in vitro experiments to detect macrophage adhesion function and stimulated with equal volumes of LPS or PBS solution for 24 h,divided into WT+PBS group,KO+PBS group,WT+LPS group and KO+LPS group,and transfected with adenovirus expressing Spry2.qRT-PCR was used to detect the mRNA expression levels of LFA-1α,IL-1β,iNOS,CD206,Arg1 and Spry2 in Mφ.Mnk1,ERK1/2,P-ERK1/2,P-p38,P-JNK and Spry2 protein levels in Mφwere detected by western blot.Results:In the in vivo experiments,the concentration of IL-1βin the serum of the KO+LPS group was more significantly elevated than that of the WT+LPS group in mice injected intraperitoneally with LPS.The expression level of splenic MφIL-1βwas higher and the mRNA expression level of Spry2 was decreased in the KO+LPS group compared to the WT+LPS group.In the in vitro experiments,the mRNA expression levels of IL-1βand iNOS were elevated and those of CD206,Arg1 and Spry2 were decreased in the KO+LPS group compared with the WT+LPS group;the expression of LFA-1αwas not significantly different in the WT+PBS and WT+LPS groups,while the expression level of LFA-1αwas significantly increased in the KO+LPS group compared with the WT+LPS group.The results of the macrophage adhesion function assay showed that the adhesion rate of Mφin the KO group was increased at several time points compared to the WT group.After LPS stimulation,the e展开更多
基金the National Natural Science Foundation of China(Grant Nos.30670954 and 30670885)
文摘We conducted a study concerning the suppressive mechanism of KAI1/CD82 on hepatoma cell metastasis.Hepatocyte growth factor(HGF)induces the migration of hepatoma cells through activation of cellular sphingosine kinase 1(SphK1).Adenovirus-mediated gene transfer of KAI1(Ad-KAI1)downregulates the SphK1 expression and suppresses the HGF-induced migration of SMMC-7721 human hepatocellcular carcinoma cells.Overexpression of KAI1/CD82 significantly elevates Sprouty2 at the protein level.Ablation of Sprouty2 with RNA interference can block the KAI1/CD82-induced suppression of hepatoma cell migration and downregulation of SphK1 expression.It is demonstrated that KAI1/CD82 suppresses HGF-induced migration of hepatoma cells via upregulation of Sprouty2.
基金supported by grants from the National Key Sci-Tech Special Project of China(2008ZX10002-018/019)the Shanghai Rising-Star Program(10QA1401300)+1 种基金the National Natural Science Foundation of China(30901432)the Research Fund for the Doctoral Program of Higher Education of China (20090071120026)
文摘BACKGROUND:In vitro experiments and mice models have confirmed the importance of Sprouty 2(Spry2) in inhibiting tumorigenesis and the progression of human cancer.However,the prognostic value of Spry2 in cancer patients remains unknown.This study is aimed to investigate the clinical relevance and prognostic significance of Spry2 expression in patients with hepatocellular carcinoma(HCC).METHODS:With samples from 240 randomly-selected HCC patients who underwent surgery,immunohistochemistry was used to investigate Spry2 expression on tissue microarrays.The correlation of Spry2 expression with survival was estimated by the Kaplan-Meier method and univariate/multivariate Cox proportional hazard regression analysis.Spry2,ERK and phospho-ERK expression in HCC cell lines was detected by Western blotting.RESULTS:Among the patients,86.3%(207 of 240) exhibited down-regulation of Spry2 expression.Patients negative for Spry2 showed poorer survival(P=0.002) and increased recurrence(P=0.003).Multivariate analysis further established Spry2 as an independent predictor of postoperative recurrence in HCC patients(HR=1.47;95% CI,1.02-2.08;P=0.037).Downregulation of Spry2 was associated with highly malignant phenotypes like vascular invasion and advanced tumor stages,and was positively correlated with the metastatic potential of HCC cell lines.CONCLUSION:In the era of molecular targeted therapy,the expression of Spry2 in HCC may have relevant clinical significance and turn out to be a key factor in prognostic assessment and in treatment planning.
基金The National Key R&D Program(2018YFC1311300)Scientific Research Project of Hubei Health Commission(WJ2021Q035)。
文摘Objective:To investigate the effect of mitogen-activated protein kinase interaction serine kinase 1(Mnk1)gene deletion on lipopolysaccharide(LPS)-induced inflammatory response in mouse macrophages(Mφ)and the possible mechanism.Methods:Healthy male wildtype C57BL/6J(WT)and Mnk1 knockout(KO)mice were selected at 8-10 weeks of age and divided into WT+PBS,KO+PBS,WT+LPS and KO+LPS groups,and the serum levels of IL-1βwere measured by ELISA after 24 h intraperitoneal injection of PBS or LPS.The mRNA expression levels of IL-1βand Sprouty2(Spry2)in the spleen Mφwere measured by qRTPCR.Mφwas also extracted from the peritoneal cavity of two strains of mice for in vitro experiments to detect macrophage adhesion function and stimulated with equal volumes of LPS or PBS solution for 24 h,divided into WT+PBS group,KO+PBS group,WT+LPS group and KO+LPS group,and transfected with adenovirus expressing Spry2.qRT-PCR was used to detect the mRNA expression levels of LFA-1α,IL-1β,iNOS,CD206,Arg1 and Spry2 in Mφ.Mnk1,ERK1/2,P-ERK1/2,P-p38,P-JNK and Spry2 protein levels in Mφwere detected by western blot.Results:In the in vivo experiments,the concentration of IL-1βin the serum of the KO+LPS group was more significantly elevated than that of the WT+LPS group in mice injected intraperitoneally with LPS.The expression level of splenic MφIL-1βwas higher and the mRNA expression level of Spry2 was decreased in the KO+LPS group compared to the WT+LPS group.In the in vitro experiments,the mRNA expression levels of IL-1βand iNOS were elevated and those of CD206,Arg1 and Spry2 were decreased in the KO+LPS group compared with the WT+LPS group;the expression of LFA-1αwas not significantly different in the WT+PBS and WT+LPS groups,while the expression level of LFA-1αwas significantly increased in the KO+LPS group compared with the WT+LPS group.The results of the macrophage adhesion function assay showed that the adhesion rate of Mφin the KO group was increased at several time points compared to the WT group.After LPS stimulation,the e
