目的探讨人乙醛脱氢酶家族1成员A3(aldehyde dehydrogenase 1 family member A3,ALDH1A3)在胰腺癌及正常胰腺组织中的表达情况,并研究其对胰腺癌侵袭的调控及可能的机制。方法利用癌症和肿瘤基因图谱(the Cancer Genome Atlas,TCGA)数...目的探讨人乙醛脱氢酶家族1成员A3(aldehyde dehydrogenase 1 family member A3,ALDH1A3)在胰腺癌及正常胰腺组织中的表达情况,并研究其对胰腺癌侵袭的调控及可能的机制。方法利用癌症和肿瘤基因图谱(the Cancer Genome Atlas,TCGA)数据库中胰腺癌患者数据集、基因组织表达项目(Genotype-Tissue Expression Project,GTEx)中正常胰腺组织基因表达谱数据和肿瘤细胞系百科全书(Cancer Cell Line Encyclopedia,CCLE)数据库中胰腺癌细胞系的基因表达谱矩阵,分析ALDH家族各亚型的表达情况;Transwell实验检测敲低ALDH1A3对胰腺癌细胞侵袭能力的影响;基因富集分析(gene set enrichment analysis,GSEA)探讨ALDH1A3与JAK/STAT3通路活化的关系。结果 ALDH1A3在胰腺癌组织中的表达高于正常胰腺组织;在胰腺癌细胞系中的表达高于其他ALDH亚型;下调ALDH1A3可以降低胰腺癌细胞系的侵袭能力;JAK/STAT3通路在ALDH1A3高表达胰腺癌患者中显著富集;下调ALDH1A3可抑制STAT3的磷酸化。结论 ALDH1A3参与调控胰腺癌细胞的侵袭,其机制可能与JAK/STAT3通路的活化有关。展开更多
目的研究氯化锂(lithium chloride,LiCl)干预对双侧卵巢切除(ovariectomized,OVX)小鼠骨丢失、Janus激酶/信号转导与转录激活子(The janus kinase/signal transducer and activator of tran-ions,JAK/STAT)信号通路及成骨细胞(osteoblas...目的研究氯化锂(lithium chloride,LiCl)干预对双侧卵巢切除(ovariectomized,OVX)小鼠骨丢失、Janus激酶/信号转导与转录激活子(The janus kinase/signal transducer and activator of tran-ions,JAK/STAT)信号通路及成骨细胞(osteoblast,OB)的影响。方法体内实验:选取8周龄雌性C57BL6/J小鼠24只,分为假手术组(Sham组),OVX组和卵巢切除+氯化锂干预(OVX+LiCl)组。收集动物左右侧股骨及右侧胫骨标本,进行Micro-CT、免疫组织化学和骨生物力学测试。体外实验:处死3日龄C57BL/6小鼠,分离颅骨培养原代OB,分为空白对照组(Control组)和LiCl组进行细胞活性检测、碱性磷酸酶染色、茜素红染色、实时荧光定量PCR检测。结果Micro-CT分析示OVX组骨量减少,骨小梁厚度变薄,骨小梁数量减少,骨小梁分离度增加,LiCl干预显著改善骨丢失。生物力学示OVX组的最大载荷、最大应力均显著低于Sham组(P<0.05),OVX+LiCl组的最大载荷、最大应力较OVX组显著升高(P<0.05)。免疫组化结果示,OVX+LiCl组JAK2和STAT3表达显著高于Sham组(P<0.05);OVX组P-STAT3表达明显低于Sham组(P<0.05),而OVX+LiCl组表达显著升高(P<0.05)。LiCl组的OB增殖活性显著高于对照组(P<0.05)。LiCl组的碱性磷酸酶活性和钙结节数量显著高于OVX组(P<0.05)。LiCl组STAT3 mRNA和OCN mRNA表达显著高于对照组(P<0.05)。结论LiCl通过调控OB改善OVX小鼠骨量和骨微结构,其机制可能与调节JAK/STAT3信号通路有关。展开更多
Recent studies regarding neuronal differentiation of mesenchymal stem cells （MSCs） have primarily focused on induction methods and transplantation in vivo. However, knowledge about the intrinsic regulatory mechanism...Recent studies regarding neuronal differentiation of mesenchymal stem cells （MSCs） have primarily focused on induction methods and transplantation in vivo. However, knowledge about the intrinsic regulatory mechanisms underlying neuronal induction of MSCs remains limited and unclear. OBJECTIVE： To elucidate the role of JAK-STAT3 signaling pathway during neuronal differentiation of MSCs using a combination of the JAK-STAT3 signaling inhibitor AG490 and growth factors. DESIGN, TIME AND SETTING： Neural, molecular, biomedical, in vitro experiment was performed at the Laboratory of Pharmacology, School of Pharmacy, Nanjing Medical University between March and December 2008 MATERIALS： An inhibitor of the JAK-STAT3 signaling pathway was purchased from Calbiochem, USA. Antibody kit for total and phosphorylated STAT3 was purchased from Cell Signaling, USA. METHODS： MSCs from passage 3 were assigned to non-induced, growth factor, and AG490 groups. MAIN OUTCOME MEASURE： The number of cells expressing neuron-specific enolase, microtubule-associated protein, and glial fibrillary acidic protein were determined by immunocytochemistry. Total and phosphorylated （Tyr705） expression levels of STAT3 protein were measured by Western blot analysis. RESULTS： MSCs were transdifferentiated into neuronal- and astrocyte-like phenotypes through the induction of epidermal growth factor, basic fibroblast growth factor, and brain-derived neurotrophic factor. In addition, the JAK-STAT3 signaling pathway was significantly activated during neural differentiation. Expression of phosphorylated （Tyr705） STAT3 was inhibited with AG490 （5 pmol/L） prior to neural induction with epidermal growth factor, basic fibroblast growth factor, and brain-derived neurotrophic factor; proportion of astrocyte-like cells was significantly reduced （P 〈 0.01）, and the proportion of neuronal-like phenotypes was significantly increased （P〈 0.01）. CONCLUSION： JAK-STAT3 signaling pathway was shown to regulate neuronal induction of b展开更多
文摘目的探讨人乙醛脱氢酶家族1成员A3(aldehyde dehydrogenase 1 family member A3,ALDH1A3)在胰腺癌及正常胰腺组织中的表达情况,并研究其对胰腺癌侵袭的调控及可能的机制。方法利用癌症和肿瘤基因图谱(the Cancer Genome Atlas,TCGA)数据库中胰腺癌患者数据集、基因组织表达项目(Genotype-Tissue Expression Project,GTEx)中正常胰腺组织基因表达谱数据和肿瘤细胞系百科全书(Cancer Cell Line Encyclopedia,CCLE)数据库中胰腺癌细胞系的基因表达谱矩阵,分析ALDH家族各亚型的表达情况;Transwell实验检测敲低ALDH1A3对胰腺癌细胞侵袭能力的影响;基因富集分析(gene set enrichment analysis,GSEA)探讨ALDH1A3与JAK/STAT3通路活化的关系。结果 ALDH1A3在胰腺癌组织中的表达高于正常胰腺组织;在胰腺癌细胞系中的表达高于其他ALDH亚型;下调ALDH1A3可以降低胰腺癌细胞系的侵袭能力;JAK/STAT3通路在ALDH1A3高表达胰腺癌患者中显著富集;下调ALDH1A3可抑制STAT3的磷酸化。结论 ALDH1A3参与调控胰腺癌细胞的侵袭,其机制可能与JAK/STAT3通路的活化有关。
基金the National Nature Science Foundation of China, No. 30973092"Xingwei" Project Medical Emphasis Grant from Jiangsu Province, No. RC2007062
文摘Recent studies regarding neuronal differentiation of mesenchymal stem cells （MSCs） have primarily focused on induction methods and transplantation in vivo. However, knowledge about the intrinsic regulatory mechanisms underlying neuronal induction of MSCs remains limited and unclear. OBJECTIVE： To elucidate the role of JAK-STAT3 signaling pathway during neuronal differentiation of MSCs using a combination of the JAK-STAT3 signaling inhibitor AG490 and growth factors. DESIGN, TIME AND SETTING： Neural, molecular, biomedical, in vitro experiment was performed at the Laboratory of Pharmacology, School of Pharmacy, Nanjing Medical University between March and December 2008 MATERIALS： An inhibitor of the JAK-STAT3 signaling pathway was purchased from Calbiochem, USA. Antibody kit for total and phosphorylated STAT3 was purchased from Cell Signaling, USA. METHODS： MSCs from passage 3 were assigned to non-induced, growth factor, and AG490 groups. MAIN OUTCOME MEASURE： The number of cells expressing neuron-specific enolase, microtubule-associated protein, and glial fibrillary acidic protein were determined by immunocytochemistry. Total and phosphorylated （Tyr705） expression levels of STAT3 protein were measured by Western blot analysis. RESULTS： MSCs were transdifferentiated into neuronal- and astrocyte-like phenotypes through the induction of epidermal growth factor, basic fibroblast growth factor, and brain-derived neurotrophic factor. In addition, the JAK-STAT3 signaling pathway was significantly activated during neural differentiation. Expression of phosphorylated （Tyr705） STAT3 was inhibited with AG490 （5 pmol/L） prior to neural induction with epidermal growth factor, basic fibroblast growth factor, and brain-derived neurotrophic factor; proportion of astrocyte-like cells was significantly reduced （P 〈 0.01）, and the proportion of neuronal-like phenotypes was significantly increased （P〈 0.01）. CONCLUSION： JAK-STAT3 signaling pathway was shown to regulate neuronal induction of b
