AIM： To study the expression of Sonic hedgehog pathway-related molecules, Sonic hedgehog （Shh） and Glil in gastric carcinoma. METHODS： Expression of Shh in 56 gastric specimens including non-cancerous gastric tiss...AIM： To study the expression of Sonic hedgehog pathway-related molecules, Sonic hedgehog （Shh） and Glil in gastric carcinoma. METHODS： Expression of Shh in 56 gastric specimens including non-cancerous gastric tissues, gastric adenocarcinoma, gastric squamous cell carcinoma was detected by RT-PCR, in situ hybridization and immunohistochemistry. Expression of Glil was observed by in situ hybridization. RESULTS： The positive rate of Shh and Glil expression was 0.0%, 0.0% in non-cancerous gastric tissues while it was 66.7%, 57.8% respectively in gastric adenocarcinoma, and 100%, 100% respectively in gastric squamous cell carcinoma. There was a significant difference between the non-cancerous gastric tissues and gastric carcinoma （P 〈 0.05）. Elevated expression of Shh and Glil in gastric tubular adenocarcinoma was associated with poorly differentiated tumors while the expression was absent in gastric mucinous adenocarcinoma. CONCLUSION： The elevated expression of Shh and Glil in gastric adenocarcinoma and gastric squamous cell carcinoma shows the involvement of activated Shh signaling in the cellular proliferation of gastric carcinogenesis. It suggests Shh signaling gene may be a new and good target gene for gastric tumor diagnosis and therapy.展开更多
AIM： To determine the role of Sonic hedgehog （Shh） pathway in colorectal adenocarcinomas through analysis of the expression of Shh pathway-related molecules, Shh, Ptchl, hedgehog-interacting protein （Hip）, Gil1, ...AIM： To determine the role of Sonic hedgehog （Shh） pathway in colorectal adenocarcinomas through analysis of the expression of Shh pathway-related molecules, Shh, Ptchl, hedgehog-interacting protein （Hip）, Gil1, Gli3 and PDGFRα. METHODS： Expression of Shh in 25 colorectal adenocarcinomas was detected by RT-PCR,in situ hybridization and immunohistochemistry. Expression of Ptchl was observed by in situ hybridization and immunohistochemistry. Expression of Hip, Gil1, Gli3 and PDGFRα was analyzed by in situ hybridization. RESULTS： Expression of cytokeratin AE1/AE3 was observed in the cytoplasm of colorectal crypts. Members of the Hh signaling pathway were expressed in colorectal epithelium. Shh was expressed in cytoplasm of dysplastic epithelial cells, while expression of Ptchl, Hip and Gill were mainly detected in the malignant crypts of adenocarcinomas. In contrast, PDGFRα was expressed highly in aberrant crypts and moderately in the stroma. Expression of Gli3 could not be detected in colorectal adenocarcinomas. CONCLUSION： These data suggest that Shh-Ptchl-Gli1 signaling pathway may play a role in the progression of colorectal tumor.展开更多
Background Anethole(AN)is an organic antioxidant compound with a benzene ring and is expected to have a positive impact on early embryogenesis in mammals.However,no study has examined the effect of AN on porcine embry...Background Anethole(AN)is an organic antioxidant compound with a benzene ring and is expected to have a positive impact on early embryogenesis in mammals.However,no study has examined the effect of AN on porcine embryonic development.Therefore,we investigated the effect of AN on the development of porcine embryos and the underlying mechanism.Results We cultured porcine in vitro-fertilized embryos in medium with AN(0,0.3,0.5,and 1 mg/mL)for 6 d.AN at 0.5 mg/mL significantly increased the blastocyst formation rate,trophectoderm cell number,and cellular survival rate compared to the control.AN-supplemented embryos exhibited significantly lower reactive oxygen species levels and higher glutathione levels than the control.Moreover,AN significantly improved the quantity of mitochondria and mitochondrial membrane potential,and increased the lipid droplet,fatty acid,and ATP levels.Interestingly,the levels of proteins and genes related to the sonic hedgehog(SHH)signaling pathway were significantly increased by AN.Conclusions These results revealed that AN improved the developmental competence of porcine preimplantation embryos by activating SHH signaling against oxidative stress and could be used for large-scale production of high-quality porcine embryos.展开更多
The effects of Sonic hedgehog(Shh) signaling pathway activation on S-type neuroblastoma(NB) cell lines and its role in NB tumorigenesis were investigated.Immunohistochemistry was used to detect the expression of Shh p...The effects of Sonic hedgehog(Shh) signaling pathway activation on S-type neuroblastoma(NB) cell lines and its role in NB tumorigenesis were investigated.Immunohistochemistry was used to detect the expression of Shh pathway components— Patched1(PTCH1) and Gli1 in 40 human primary NB samples.Western blotting and RT-PCR were used to examine the protein expression and mRNA levels of PTCH1 and Gli1 in three kinds of S-type NB cell lines(SK-N-AS,SK-N-SH and SHEP1),respectively.Exogenous Shh was administrated to activate Shh signaling pathway while cyclopamine was used as a selective antagonist of Shh pathway.S-type NB cell lines were treated with different concentrations of Shh or/and cyclopamine for different durations.Cell viability was measured by using MTT method.Apoptosis rate and cell cycle were assayed by flow cytometry.The xenograft experiments were used to evaluate the role of Shh pathway in tumor growth in immunodeficient mice.High-level expression of PTCH1 and Gli1 was detected in both NB samples and S-type NB cell lines.Cyclopamine decreased the survival rate of the three cell lines while Shh increased it,and the inhibition effects of cyclopamine could be partially reversed by shh pre-treatment.Cyclopamine induced the cell apoptosis and the cell cycle arrest in G0/G1 phase,while Shh induced the reverse effects and could partially prevent effects of cyclopamine.Cyclopamine could also inhibit the growth of NB in vivo.Our studies revealed that activation of the Shh pathway is important for survival and proliferation of S-type NB cells in vivo and in vitro through affecting cell apoptosis and cell cycle,suggesting a new therapeutic approach to NB.展开更多
Proliferation of neural stem cells is regulated by the secreted signaling molecule sonic hedgehog. In this study, neural stem cells were infected with recombinant adeno-associated virus expressing sonic hedgehog-N-enh...Proliferation of neural stem cells is regulated by the secreted signaling molecule sonic hedgehog. In this study, neural stem cells were infected with recombinant adeno-associated virus expressing sonic hedgehog-N-enhanced green fluorescent protein. The results showed that overexpression of sonic hedgehog in neural stem cells induced the increased expression of Gill and N-myc, a target gene of sonic hedgehog. These findings suggest that N-myc is a direct downstream target of the sonic hedgehog signal pathway in neural stem cells. Sonic hedgehog and N-myc are important mediators of sonic hedgehog-induced proliferation of neural stem cells.展开更多
Background:The sonic hedgehog(SHH)pathway is an important signaling pathway for neural tube closure.GLI family zinc finger 2(GLI2)is the major activation mediator of the SHH pathway;however,no single-nucleotide polymo...Background:The sonic hedgehog(SHH)pathway is an important signaling pathway for neural tube closure.GLI family zinc finger 2(GLI2)is the major activation mediator of the SHH pathway;however,no single-nucleotide polymorphisms(SNPs)in GLI2 have been reported to be associated with human neural tube defects(NTDs)to date.Here,we evaluated a mutation in GLI2 in the Han Chinese population.Methods:We used SNPscan to genotype rs3738880 in the GLI2 coding region.We then investigated the function of this gene by Western blotting and dual-luciferase assays.Results:In this study,we found that the GLI2 missense variant rs3738880 significantly increased the risk of NTDs in the Han Chinese population via association studies in a cohort of 254 patients and 277 controls from Shanxi Province(odds ratio[OR]=1.89,95%confidence interval[CI]=1.28-2.80,P=0.0012).Additional stratified analyses demonstrated that rs3738880 was significantly related to spina bifida(114 cases,OR=2.01,95%CI=1.19–3.38,P=0.0067).Functional analysis revealed that rs3738880 did not affect GLI2 protein stability and significantly increased SHH activity because of the introduction of a potential phosphorylation site in GLI2.Conclusion:rs3738880 was a risk factor for NTDs in the Han Chinese population.展开更多
Aim:Hepatocellular carcinoma(HCC)is one of the leading causes of cancer-associated death.The Sonic Hedgehog(SHH)signaling pathway participates in the initiation,progression,migration,and recurrence of HCC cancer stem ...Aim:Hepatocellular carcinoma(HCC)is one of the leading causes of cancer-associated death.The Sonic Hedgehog(SHH)signaling pathway participates in the initiation,progression,migration,and recurrence of HCC cancer stem cells.Furthermore,SHH regulates various cellular behaviors such as proliferation,differentiation,survival,self-renewal,epithelial-mesenchymal transition(EMT),and SHH autoregulation.Glioma-associated oncogene(GLI)family zinc finger are key transcription factors in the development of many organs and are deregulated in cancer.In this study,Huh-7 cells were treated with GLI-specific decoy oligodeoxynucleotide(ODN)to evaluate its anticancer impact.Methods:The transfection efficiency of GLI-specific decoy ODN was measured using fluorescent microscopy.Then,the effects of GLI-specific decoy ODN on apoptosis,viability,proliferation rate,colony formation,and migration capacities of Huh-7 cells were assessed.Furthermore,the expression of genes associated with the alteration of SHH was assessed.Results:Treatment of Huh-7 cells with GLI-specific decoy ODN decreased cell viability(56.36%±3%).Expression of certain genes such as c-MYC,SNAI2,ZEB1,and PROM1 decreased dramatically,while the expression of CDH1 increased significantly.Furthermore,the treated cells’proliferation,colony formation,and migration capacity decreased considerably.This treatment induced apoptosis in the Huh-7 cells.Conclusion:Inhibition of the SHH signaling pathway using GLI-specific decoy ODN led to a decline in the growth rate of HCC cells,decreased migration,and attenuated EMT progression.展开更多
基金Supported by the Foundation of Shandong Province Bureau of Health, No. 2005JZ001
文摘AIM： To study the expression of Sonic hedgehog pathway-related molecules, Sonic hedgehog （Shh） and Glil in gastric carcinoma. METHODS： Expression of Shh in 56 gastric specimens including non-cancerous gastric tissues, gastric adenocarcinoma, gastric squamous cell carcinoma was detected by RT-PCR, in situ hybridization and immunohistochemistry. Expression of Glil was observed by in situ hybridization. RESULTS： The positive rate of Shh and Glil expression was 0.0%, 0.0% in non-cancerous gastric tissues while it was 66.7%, 57.8% respectively in gastric adenocarcinoma, and 100%, 100% respectively in gastric squamous cell carcinoma. There was a significant difference between the non-cancerous gastric tissues and gastric carcinoma （P 〈 0.05）. Elevated expression of Shh and Glil in gastric tubular adenocarcinoma was associated with poorly differentiated tumors while the expression was absent in gastric mucinous adenocarcinoma. CONCLUSION： The elevated expression of Shh and Glil in gastric adenocarcinoma and gastric squamous cell carcinoma shows the involvement of activated Shh signaling in the cellular proliferation of gastric carcinogenesis. It suggests Shh signaling gene may be a new and good target gene for gastric tumor diagnosis and therapy.
基金grants from National Natural Science Foundation of China, No. 30228031, No. 30671072
文摘AIM： To determine the role of Sonic hedgehog （Shh） pathway in colorectal adenocarcinomas through analysis of the expression of Shh pathway-related molecules, Shh, Ptchl, hedgehog-interacting protein （Hip）, Gil1, Gli3 and PDGFRα. METHODS： Expression of Shh in 25 colorectal adenocarcinomas was detected by RT-PCR,in situ hybridization and immunohistochemistry. Expression of Ptchl was observed by in situ hybridization and immunohistochemistry. Expression of Hip, Gil1, Gli3 and PDGFRα was analyzed by in situ hybridization. RESULTS： Expression of cytokeratin AE1/AE3 was observed in the cytoplasm of colorectal crypts. Members of the Hh signaling pathway were expressed in colorectal epithelium. Shh was expressed in cytoplasm of dysplastic epithelial cells, while expression of Ptchl, Hip and Gill were mainly detected in the malignant crypts of adenocarcinomas. In contrast, PDGFRα was expressed highly in aberrant crypts and moderately in the stroma. Expression of Gli3 could not be detected in colorectal adenocarcinomas. CONCLUSION： These data suggest that Shh-Ptchl-Gli1 signaling pathway may play a role in the progression of colorectal tumor.
基金supported by the Ministry of EducationScience and Technology(No.2021M3A9A1096894)+1 种基金Republic of Korea and the KRIBB Research Initiative Program(KGM4252223)Korea Research Institute of Bioscience and Biotechnology,Republic of Korea。
文摘Background Anethole(AN)is an organic antioxidant compound with a benzene ring and is expected to have a positive impact on early embryogenesis in mammals.However,no study has examined the effect of AN on porcine embryonic development.Therefore,we investigated the effect of AN on the development of porcine embryos and the underlying mechanism.Results We cultured porcine in vitro-fertilized embryos in medium with AN(0,0.3,0.5,and 1 mg/mL)for 6 d.AN at 0.5 mg/mL significantly increased the blastocyst formation rate,trophectoderm cell number,and cellular survival rate compared to the control.AN-supplemented embryos exhibited significantly lower reactive oxygen species levels and higher glutathione levels than the control.Moreover,AN significantly improved the quantity of mitochondria and mitochondrial membrane potential,and increased the lipid droplet,fatty acid,and ATP levels.Interestingly,the levels of proteins and genes related to the sonic hedgehog(SHH)signaling pathway were significantly increased by AN.Conclusions These results revealed that AN improved the developmental competence of porcine preimplantation embryos by activating SHH signaling against oxidative stress and could be used for large-scale production of high-quality porcine embryos.
基金supported by a grant from the National Natural Sciences Foundation of China (No.30600189)
文摘The effects of Sonic hedgehog(Shh) signaling pathway activation on S-type neuroblastoma(NB) cell lines and its role in NB tumorigenesis were investigated.Immunohistochemistry was used to detect the expression of Shh pathway components— Patched1(PTCH1) and Gli1 in 40 human primary NB samples.Western blotting and RT-PCR were used to examine the protein expression and mRNA levels of PTCH1 and Gli1 in three kinds of S-type NB cell lines(SK-N-AS,SK-N-SH and SHEP1),respectively.Exogenous Shh was administrated to activate Shh signaling pathway while cyclopamine was used as a selective antagonist of Shh pathway.S-type NB cell lines were treated with different concentrations of Shh or/and cyclopamine for different durations.Cell viability was measured by using MTT method.Apoptosis rate and cell cycle were assayed by flow cytometry.The xenograft experiments were used to evaluate the role of Shh pathway in tumor growth in immunodeficient mice.High-level expression of PTCH1 and Gli1 was detected in both NB samples and S-type NB cell lines.Cyclopamine decreased the survival rate of the three cell lines while Shh increased it,and the inhibition effects of cyclopamine could be partially reversed by shh pre-treatment.Cyclopamine induced the cell apoptosis and the cell cycle arrest in G0/G1 phase,while Shh induced the reverse effects and could partially prevent effects of cyclopamine.Cyclopamine could also inhibit the growth of NB in vivo.Our studies revealed that activation of the Shh pathway is important for survival and proliferation of S-type NB cells in vivo and in vitro through affecting cell apoptosis and cell cycle,suggesting a new therapeutic approach to NB.
基金funded by the National Natural Science Foundation of China,No.81171401Science and Technology Development Program of Dalian City,No.2008J99JH268the Scientific Research Program of Higher Learning School of Department of Education of Liaoning Province,No.L20100108
文摘Proliferation of neural stem cells is regulated by the secreted signaling molecule sonic hedgehog. In this study, neural stem cells were infected with recombinant adeno-associated virus expressing sonic hedgehog-N-enhanced green fluorescent protein. The results showed that overexpression of sonic hedgehog in neural stem cells induced the increased expression of Gill and N-myc, a target gene of sonic hedgehog. These findings suggest that N-myc is a direct downstream target of the sonic hedgehog signal pathway in neural stem cells. Sonic hedgehog and N-myc are important mediators of sonic hedgehog-induced proliferation of neural stem cells.
基金This work was supported by grants from the National Key Basic Research Program of China[2016YFC1000502 to H.Wang and X Yang]the National Natural Science Foundation of China[81430005 to H Wang,81472050 to X Yang]Open project of National Population and Family Planning Key Laboratory of Contraceptive drugs and Devices[2016KF04 for X Yang].
文摘Background:The sonic hedgehog(SHH)pathway is an important signaling pathway for neural tube closure.GLI family zinc finger 2(GLI2)is the major activation mediator of the SHH pathway;however,no single-nucleotide polymorphisms(SNPs)in GLI2 have been reported to be associated with human neural tube defects(NTDs)to date.Here,we evaluated a mutation in GLI2 in the Han Chinese population.Methods:We used SNPscan to genotype rs3738880 in the GLI2 coding region.We then investigated the function of this gene by Western blotting and dual-luciferase assays.Results:In this study,we found that the GLI2 missense variant rs3738880 significantly increased the risk of NTDs in the Han Chinese population via association studies in a cohort of 254 patients and 277 controls from Shanxi Province(odds ratio[OR]=1.89,95%confidence interval[CI]=1.28-2.80,P=0.0012).Additional stratified analyses demonstrated that rs3738880 was significantly related to spina bifida(114 cases,OR=2.01,95%CI=1.19–3.38,P=0.0067).Functional analysis revealed that rs3738880 did not affect GLI2 protein stability and significantly increased SHH activity because of the introduction of a potential phosphorylation site in GLI2.Conclusion:rs3738880 was a risk factor for NTDs in the Han Chinese population.
基金supported by grants from the Royan Institute(97000205)Bahar Tashkhis Teb Co.(BTT,9703,9809,and 9903).
文摘Aim:Hepatocellular carcinoma(HCC)is one of the leading causes of cancer-associated death.The Sonic Hedgehog(SHH)signaling pathway participates in the initiation,progression,migration,and recurrence of HCC cancer stem cells.Furthermore,SHH regulates various cellular behaviors such as proliferation,differentiation,survival,self-renewal,epithelial-mesenchymal transition(EMT),and SHH autoregulation.Glioma-associated oncogene(GLI)family zinc finger are key transcription factors in the development of many organs and are deregulated in cancer.In this study,Huh-7 cells were treated with GLI-specific decoy oligodeoxynucleotide(ODN)to evaluate its anticancer impact.Methods:The transfection efficiency of GLI-specific decoy ODN was measured using fluorescent microscopy.Then,the effects of GLI-specific decoy ODN on apoptosis,viability,proliferation rate,colony formation,and migration capacities of Huh-7 cells were assessed.Furthermore,the expression of genes associated with the alteration of SHH was assessed.Results:Treatment of Huh-7 cells with GLI-specific decoy ODN decreased cell viability(56.36%±3%).Expression of certain genes such as c-MYC,SNAI2,ZEB1,and PROM1 decreased dramatically,while the expression of CDH1 increased significantly.Furthermore,the treated cells’proliferation,colony formation,and migration capacity decreased considerably.This treatment induced apoptosis in the Huh-7 cells.Conclusion:Inhibition of the SHH signaling pathway using GLI-specific decoy ODN led to a decline in the growth rate of HCC cells,decreased migration,and attenuated EMT progression.
