Bone morphogenetic proteins (BMPs) are members of the TGF-β superfamily and have diverse functions during development and organogenesis. BMPs play a major role in skeletal development and bone formation, and disrupti...Bone morphogenetic proteins (BMPs) are members of the TGF-β superfamily and have diverse functions during development and organogenesis. BMPs play a major role in skeletal development and bone formation, and disruptions in BMP signaling cause a variety of skeletal and extraskeletal anomalies. Several knockout models have provided insight into the mechanisms responsible for these phenotypes. Proper bone formation requires the differentiation of osteoblasts from mesenchymal stem cell (MSC) precursors, a process mediated in part by BMP signaling. Multiple BMPs, including BMP2, BMP6, BMP7 and BMP9, promote osteoblastic differentiation of MSCs both in vitro and in vivo. BMP9 is one of the most osteogenic BMPs, yet it is a poorly characterized member of the BMP family. Several studies demonstrate that the mechanisms controlling BMP9-mediated osteogenesis differ from other osteogenic BMPs, but little is known about these specific mechanisms. Several pathways critical to BMP9-mediated osteogenesis are also important in the differentiation of other cell lineages, including adipocytes and chondrocytes. BMP9 has also demonstrated translational promise in spinal fusion and bone fracture repair. This review will summarize our current knowledge of BMP-mediated osteogenesis, with a focus on BMP9, by presenting recently completed work which may help us to further elucidate these pathways.展开更多
目的观察脓毒症相关急性呼吸窘迫综合征(acute respiratory distress syndrome,ARDS)患者骨形态发生蛋白9(bone morphogenetic protein 9,BMP9)表达水平,探讨BMP9在脓毒症相关ARDS早期识别及预后预测中的作用。方法选取山西白求恩医院2...目的观察脓毒症相关急性呼吸窘迫综合征(acute respiratory distress syndrome,ARDS)患者骨形态发生蛋白9(bone morphogenetic protein 9,BMP9)表达水平,探讨BMP9在脓毒症相关ARDS早期识别及预后预测中的作用。方法选取山西白求恩医院2022年5月至2023年5月收治的脓毒症相关ARDS患者56例作为ARDS组,心源性肺水肿患者49例作为病例对照组,同期于本院体检中心进行健康体检的成人46名作为健康对照组。追踪随访ARDS组患者28 d的转归情况,分为存活组26例和死亡组30例。分析比较各组受试者血清BMP9表达水平及其与临床指标的相关性,采用Logistic回归分析脓毒症相关ARDS发病的危险因素,并对相关指标进行诊断效能及预后预测价值分析。结果脓毒症相关ARDS组患者血清BMP9水平[1401.14(856.59,1982.86)pg/mL]高于病例对照组(438.26±128.52)pg/mL及健康对照组(398.96±96.55)pg/mL,差异有统计学意义(P<0.01)。而且BMP9表达与炎症指标降钙素原、淋巴细胞计数及疾病严重程度序贯器官衰竭评分(sequential organ failure assessment,SOFA)均显著相关(P<0.05,P<0.01)。采用多因素Logistic回归分析显示,BMP9是脓毒症相关ARDS发病的高危因素(P<0.01)。绘制受试者工作特征曲线(receiver operating characteristic curve,ROC),BMP9预测脓毒症相关ARDS发生的ROC曲线下面积(area under the ROC curve,AUC)为0.930,特异度为100.0%,敏感度为80.4%,明显高于氧合指数的特异度(89.8%)和敏感度(67.9%)。追踪随访并比较脓毒症相关ARDS不同预后患者BMP9水平,发现相较于存活组,死亡组患者BMP9表达水平更高,差异有统计学意义(P<0.05)。绘制ROC曲线,分析BMP9在脓毒症相关ARDS患者预后预测中的作用,ROC曲线下面积为0.699,敏感度为43.3%,特异度为100.0%。结论脓毒症相关ARDS患者BMP9表达明显升高,而且其高表达与炎症指标降钙素原、淋巴细胞计数及疾病严重SOFA评分呈显著相关性。BMP9是脓毒症相关展开更多
Mesenchymal stem cells(MSCs)are multipotent stem cells and capable of differentiating into multiple cell types including osteoblastic,chondrogenic and adipogenic lineages.We previously identified BMP9 as one of the mo...Mesenchymal stem cells(MSCs)are multipotent stem cells and capable of differentiating into multiple cell types including osteoblastic,chondrogenic and adipogenic lineages.We previously identified BMP9 as one of the most potent BMPs that induce osteoblastic differentiation of MSCs although exact molecular mechanism through which BMP9 regulates osteogenic differentiation remains to be fully understood.Here,we seek to develop a recombinant adenovirus system to optimally silence mouse BMP9 and then characterize the important role of BMP9 in osteogenic differentiation of MSCs.Using two different siRNA bioinformatic prediction programs,we design five siRNAs targeting mouse BMP9(or simB9),which are expressed under the control of the converging H1 and U6 promoters in recombinant adenovirus vectors.We demonstrate that two of the five siRNAs,simB9-4 and simB9-7,exhibit the highest efficiency on silencing exogenous mouse BMP9 in MSCs.Furthermore,simB9-4 and simB9-7 act synergistically in inhibiting BMP9-induced expression of osteogenic markers,matrix mineralization and ectopic bone formation from MSCs.Thus,our findings demonstrate the important role of BMP9 in osteogenic differentiation of MSCs.The characterized simB9 siRNAs may be used as an important tool to investigate the molecular mechanism behind BMP9 osteogenic signaling.Our results also indicate that recombinant adenovirus-mediated expression of siRNAs is efficient and sustained,and thus may be used as an effective delivery vehicle of siRNA therapeutics.展开更多
Bone defects and non-union are prevalent in clinical orthopedy,and the outcomes of current treatments are often suboptimal.Bone tissue engineering offers a promising approach to treating these conditions effectively.B...Bone defects and non-union are prevalent in clinical orthopedy,and the outcomes of current treatments are often suboptimal.Bone tissue engineering offers a promising approach to treating these conditions effectively.Bone morphogenetic protein 9(BMP9)can commit mesenchymal stem cells to osteogenic lineage,and a knowledge of the underlying mechanisms may help advance the field of bone tissue engineering.Leucine-rich repeats con-taining G protein-coupled receptor 4(LGR4),a member of G protein-coupled receptors,is essential for modulating bone development.This study is aimed at investigating the impact of LGR4 on BMP9-induced osteogenesis in mesenchymal stem cells as well as the underlying mechanisms.Bone marrow stromal cells from BMp9-knockout mice exhibited diminished LGR4 expression,and exogenous LGR4 clearly restored the impaired osteogenic potency of the bone marrow stromal cells.Furthermore,LGR4 expression was increased by BMP9 in C3H10T1/2 cells.LGR4 augmented the benefits of BMP9-induced osteogenic markers and bone formation,whereas LGR4 inhibition restricted these effects.Meanwhile,the BMP9-induced li-pogenic markers were increased by LGR4 inhibition.The protein levels of Raptor and p-Stat3 were elevated by BMP9.Raptor knockdown or p-Stat3 suppression attenuated the osteoblastic markers and LGR4 expression brought on by BMP9.LGR4 significantly reversed the blocking ef-fect of Raptor knockdown or p-Stat3 suppression on the BMP9-induced osteoblastic markers.Raptor interacts with p-Stat3,and p-Stat3 activates the LGR4 promoter activity.In conclusion,LGR4 boosts BMP9 osteoblastic potency in mesenchymal stem cells,and BMP9 may up-regulate LGR4 via the mTORC1/Stat3 signal activation.展开更多
动脉性肺动脉高压(Pulmonary Arterial Hypertension, PAH),即第1大类肺动脉高压,是一种以高发病率和死亡率为特点的血管疾病,其主要表现是肺血管重塑和肺血管阻力增加,最终导致右心室衰竭甚至死亡。骨形态发生蛋白9 (Bone Morphogeneti...动脉性肺动脉高压(Pulmonary Arterial Hypertension, PAH),即第1大类肺动脉高压,是一种以高发病率和死亡率为特点的血管疾病,其主要表现是肺血管重塑和肺血管阻力增加,最终导致右心室衰竭甚至死亡。骨形态发生蛋白9 (Bone Morphogenetic Protein 9, BMP9)属于转化生长因子β (Transforming Growth Factor-β, TGF-β)家族,主要由肝脏星状细胞产生,随血液循环到肺血管内皮细胞上与受体结合,在PAH中发挥相应的生物学效应,但是部分研究结果是相互矛盾的。本文对PAH中BMP9的信号通路及BMP9对肺血管内皮细胞作用的研究和进展进行综述。Arterial pulmonary hypertension (PAH), also known as the group 1 pulmonary hypertension, is a vascular disease characterized by high morbidity and mortality, and its main manifestations are pulmonary vascular remodeling and increased pulmonary vascular resistance, which eventually leads to right ventricular failure and even death. Bone Morphogenetic Protein 9 (BMP9) belongs to the Transforming Growth Factor-β (TGF-β) family, which is mainly produced by hepatic stellate cells, which circulates to pulmonary vascular endothelial cells with blood and bind to their receptors to exert corresponding biological effects in PAH, but some research results are contradictory. This article briefly reviews the BMP9 signaling pathway in PAH and the research and progress of BMP9 on pulmonary vascular endothelial cells.展开更多
Silk as a natural biomaterial is considered as a promising bone substitute in tissue regeneration.Sericin and fibroin are the main components of silk and display unique features for their programmable mechanical prope...Silk as a natural biomaterial is considered as a promising bone substitute in tissue regeneration.Sericin and fibroin are the main components of silk and display unique features for their programmable mechanical properties,biocompatibility,biodegradability and morphological plasticity.It has been reported that sericin recombinant growth factors(GFs)can support cell proliferation and induce stem cell differentiation through cross-talk of signaling pathways during tissue regeneration.The transgenic technology allows the productions of bioactive heterologous GFs as fusion proteins with sericin,which are then fabricated into solid matrix or hydrogel format.Herein,using an injectable hydrogel derived from transgenic platelet-derived GF(PDGF)-BB silk sericin,we demonstrated that the PDGF-BB sericin hydrogel effectively augmented osteogenesis induced by bone morphogenetic protein(BMP9)-stimulated mesenchymal stem cells(MSCs)in vivo and in vitro,while inhibiting adipogenic differentiation.Further gene expression and protein-protein interactions studies demonstrated that BMP9 and PDGF-BB synergistically induced osteogenic differentiation through the cross-talk between Smad and Stat3 pathways in MSCs.Thus,our results provide a novel strategy to encapsulate osteogenic factors and osteoblastic progenitors in transgenic sericin-based hydrogel for robust bone tissue engineering.展开更多
目的:研究骨形态发生蛋白9(Bone morphogenetic protein 9,BMP9)对乳腺癌细胞MDA-MB-231增殖、周期及凋亡的影响。方法:半定量RT-PCR法检测高转移性乳腺癌细胞MDA-MB-231和低转移性乳腺癌细胞MCF-7中BMP9的表达,用人BMP9重组腺病毒(AdBM...目的:研究骨形态发生蛋白9(Bone morphogenetic protein 9,BMP9)对乳腺癌细胞MDA-MB-231增殖、周期及凋亡的影响。方法:半定量RT-PCR法检测高转移性乳腺癌细胞MDA-MB-231和低转移性乳腺癌细胞MCF-7中BMP9的表达,用人BMP9重组腺病毒(AdBMP9)感染MDA-MB-231细胞作为实验组,含GFP的空载腺病毒(AdGFP)感染MDA-MB-231细胞作为对照组,MTT法观察细胞增殖的变化,流式细胞仪分析周期及凋亡的变化。结果:半定量RT-PCR显示,乳腺癌细胞MDA-MB-231中未检测到BMP9 mRNA的表达;MTT结果显示,AdBMP9处理可引起MDA-MB-231细胞增殖抑制,第5 d MDA-MB-231/BMP9组的细胞增殖率(0.8860±0.0532)显著低于MDA-MB-231/GFP组(1.2240±0.1031)(P<0.05);流式细胞仪分析结果显示,腺病毒感染后第2 d和第3 d,MDA-MB-231/BMP9组细胞周期各相发生变化,G2/M期细胞明显增加,分别为MDA-MB-231/GFP组的3.2倍和2.4倍(P<0.05);同时BMP9可诱发细胞凋亡,腺病毒感染后第3 d MDA-MB-231/BMP9组细胞凋亡百分率(31.55%±8.26%)明显高于MDA-MB-231/GFP组(3.80%±0.46%)(P<0.05)。结论:提示BMP9可通过阻滞细胞周期进程和诱导细胞凋亡来抑制乳腺癌细胞MDA-MB-231的增殖。展开更多
BMP9 mediated osteogenic differentiation mechanisms of MSCs were widely explored, however, mechanisms of BMP9-induced angiogenesis still need to be clarified. We previously characterized that Notch1 promoted BMP9-indu...BMP9 mediated osteogenic differentiation mechanisms of MSCs were widely explored, however, mechanisms of BMP9-induced angiogenesis still need to be clarified. We previously characterized that Notch1 promoted BMP9-induced osteogenesis–angiogenesis coupling process in mesenchymal stem cells (MSCs). Here, we explored the underlying mechanisms of lncRNA H19 (H19) mediated regulation of BMP9-induced angiogenesis through activating Notch1 signaling. We demonstrated that basal expression level of H19 was high in MSCs, and silencing H19 attenuates BMP9-induced osteogenesis and angiogenesis of MSCs both in vitro and in vivo. Meanwhile, we identified that BMP9-induced production of CD31+ cells was indispensable for BMP9-induced bone formation, and silencing H19 dramatically blocked BMP9-induced production of CD31^(+) cells. In addition, we found that down-regulation of H19 inhibited BMP9 mediated blood vessel formation and followed subsequent bone formation in vivo. Mechanistically, we clarified that H19 promoted p53 phosphorylation by direct interacting and phosphorylating binding, and phosphorylated p53 potentiated Notch1 expression and activation of Notch1 targeting genes by binding on the promoter area of Notch1 gene. These findings suggested that H19 regulated BMP9-induced angiogenesis of MSCs by promoting the p53-Notch1 angiogenic signaling axis.展开更多
Despite advances in screening and treatment,colon cancer remains one of the leading causes of cancer-related death.Finding novel and useful drug treatment targets is also an urgent need for clinical applications.Tetra...Despite advances in screening and treatment,colon cancer remains one of the leading causes of cancer-related death.Finding novel and useful drug treatment targets is also an urgent need for clinical applications.Tetrandrine(Tet)is extracted from the Chinese medicinal herbal medicine,which is a well-known calcium blocker with a variety of pharmacological activities,including anti-cancer.In this study,we recruited cell viability assay,flow cytometry analysis,cloning formation to confirm that Tet can inhibit the proliferation of SW620 cells,and induce apoptosis.Mechanically,we confirmed that Tet up-regulates the mRNA and protein level of BMP9 in SW620 cells.Over-expression BMP9 enhances the anticancer effects of Tet in SW620 cells,but these effects can be partly reversed by silencing BMP9.Also,Tet reduces phosphorylation of Aktl/2/3 in SW620 cells,which could be elevated by overexpressed BMP9 and impaired by silencing BMP9.Furthermore,we demonstrated that Tet reduces phosphorylated PTEN,which can be promoted by overexpressed BMP9,analogously also be attenuated through silencing BMP9.Finally,we introduced a xenograft tumor model to investigate the anti-proliferative effect of Tet,further to explore the effects of BMP9 and PTEN in SW620 cells.Our findings suggested that the anti-cancer activity of Tet in SW620 cells may be mediated partly by up-regulating BMP9,followed by inactivation PI3K/Akt through up-regulating PTEN at least.展开更多
文摘Bone morphogenetic proteins (BMPs) are members of the TGF-β superfamily and have diverse functions during development and organogenesis. BMPs play a major role in skeletal development and bone formation, and disruptions in BMP signaling cause a variety of skeletal and extraskeletal anomalies. Several knockout models have provided insight into the mechanisms responsible for these phenotypes. Proper bone formation requires the differentiation of osteoblasts from mesenchymal stem cell (MSC) precursors, a process mediated in part by BMP signaling. Multiple BMPs, including BMP2, BMP6, BMP7 and BMP9, promote osteoblastic differentiation of MSCs both in vitro and in vivo. BMP9 is one of the most osteogenic BMPs, yet it is a poorly characterized member of the BMP family. Several studies demonstrate that the mechanisms controlling BMP9-mediated osteogenesis differ from other osteogenic BMPs, but little is known about these specific mechanisms. Several pathways critical to BMP9-mediated osteogenesis are also important in the differentiation of other cell lineages, including adipocytes and chondrocytes. BMP9 has also demonstrated translational promise in spinal fusion and bone fracture repair. This review will summarize our current knowledge of BMP-mediated osteogenesis, with a focus on BMP9, by presenting recently completed work which may help us to further elucidate these pathways.
文摘目的观察脓毒症相关急性呼吸窘迫综合征(acute respiratory distress syndrome,ARDS)患者骨形态发生蛋白9(bone morphogenetic protein 9,BMP9)表达水平,探讨BMP9在脓毒症相关ARDS早期识别及预后预测中的作用。方法选取山西白求恩医院2022年5月至2023年5月收治的脓毒症相关ARDS患者56例作为ARDS组,心源性肺水肿患者49例作为病例对照组,同期于本院体检中心进行健康体检的成人46名作为健康对照组。追踪随访ARDS组患者28 d的转归情况,分为存活组26例和死亡组30例。分析比较各组受试者血清BMP9表达水平及其与临床指标的相关性,采用Logistic回归分析脓毒症相关ARDS发病的危险因素,并对相关指标进行诊断效能及预后预测价值分析。结果脓毒症相关ARDS组患者血清BMP9水平[1401.14(856.59,1982.86)pg/mL]高于病例对照组(438.26±128.52)pg/mL及健康对照组(398.96±96.55)pg/mL,差异有统计学意义(P<0.01)。而且BMP9表达与炎症指标降钙素原、淋巴细胞计数及疾病严重程度序贯器官衰竭评分(sequential organ failure assessment,SOFA)均显著相关(P<0.05,P<0.01)。采用多因素Logistic回归分析显示,BMP9是脓毒症相关ARDS发病的高危因素(P<0.01)。绘制受试者工作特征曲线(receiver operating characteristic curve,ROC),BMP9预测脓毒症相关ARDS发生的ROC曲线下面积(area under the ROC curve,AUC)为0.930,特异度为100.0%,敏感度为80.4%,明显高于氧合指数的特异度(89.8%)和敏感度(67.9%)。追踪随访并比较脓毒症相关ARDS不同预后患者BMP9水平,发现相较于存活组,死亡组患者BMP9表达水平更高,差异有统计学意义(P<0.05)。绘制ROC曲线,分析BMP9在脓毒症相关ARDS患者预后预测中的作用,ROC曲线下面积为0.699,敏感度为43.3%,特异度为100.0%。结论脓毒症相关ARDS患者BMP9表达明显升高,而且其高表达与炎症指标降钙素原、淋巴细胞计数及疾病严重SOFA评分呈显著相关性。BMP9是脓毒症相关
基金The reported work was supported in part by research grants from the National Institutes of Health(CA226303,DE020140 to TCH and RRR)the U.S.Department of Defense(OR130096 to JMW)+1 种基金the Chicago Biomedical Consortium with support from the Searle Funds at The Chicago Community Trust(RRR,TCH),the Scoliosis Research Society(TCH and MJL)the National Key Research and Development Program of China(2016YFC1000803 and 2011CB707906 to TCH).
文摘Mesenchymal stem cells(MSCs)are multipotent stem cells and capable of differentiating into multiple cell types including osteoblastic,chondrogenic and adipogenic lineages.We previously identified BMP9 as one of the most potent BMPs that induce osteoblastic differentiation of MSCs although exact molecular mechanism through which BMP9 regulates osteogenic differentiation remains to be fully understood.Here,we seek to develop a recombinant adenovirus system to optimally silence mouse BMP9 and then characterize the important role of BMP9 in osteogenic differentiation of MSCs.Using two different siRNA bioinformatic prediction programs,we design five siRNAs targeting mouse BMP9(or simB9),which are expressed under the control of the converging H1 and U6 promoters in recombinant adenovirus vectors.We demonstrate that two of the five siRNAs,simB9-4 and simB9-7,exhibit the highest efficiency on silencing exogenous mouse BMP9 in MSCs.Furthermore,simB9-4 and simB9-7 act synergistically in inhibiting BMP9-induced expression of osteogenic markers,matrix mineralization and ectopic bone formation from MSCs.Thus,our findings demonstrate the important role of BMP9 in osteogenic differentiation of MSCs.The characterized simB9 siRNAs may be used as an important tool to investigate the molecular mechanism behind BMP9 osteogenic signaling.Our results also indicate that recombinant adenovirus-mediated expression of siRNAs is efficient and sustained,and thus may be used as an effective delivery vehicle of siRNA therapeutics.
基金All animal experiments were approved by The Ethics Committee of Chongqing Medical University(No.2022030).
文摘Bone defects and non-union are prevalent in clinical orthopedy,and the outcomes of current treatments are often suboptimal.Bone tissue engineering offers a promising approach to treating these conditions effectively.Bone morphogenetic protein 9(BMP9)can commit mesenchymal stem cells to osteogenic lineage,and a knowledge of the underlying mechanisms may help advance the field of bone tissue engineering.Leucine-rich repeats con-taining G protein-coupled receptor 4(LGR4),a member of G protein-coupled receptors,is essential for modulating bone development.This study is aimed at investigating the impact of LGR4 on BMP9-induced osteogenesis in mesenchymal stem cells as well as the underlying mechanisms.Bone marrow stromal cells from BMp9-knockout mice exhibited diminished LGR4 expression,and exogenous LGR4 clearly restored the impaired osteogenic potency of the bone marrow stromal cells.Furthermore,LGR4 expression was increased by BMP9 in C3H10T1/2 cells.LGR4 augmented the benefits of BMP9-induced osteogenic markers and bone formation,whereas LGR4 inhibition restricted these effects.Meanwhile,the BMP9-induced li-pogenic markers were increased by LGR4 inhibition.The protein levels of Raptor and p-Stat3 were elevated by BMP9.Raptor knockdown or p-Stat3 suppression attenuated the osteoblastic markers and LGR4 expression brought on by BMP9.LGR4 significantly reversed the blocking ef-fect of Raptor knockdown or p-Stat3 suppression on the BMP9-induced osteoblastic markers.Raptor interacts with p-Stat3,and p-Stat3 activates the LGR4 promoter activity.In conclusion,LGR4 boosts BMP9 osteoblastic potency in mesenchymal stem cells,and BMP9 may up-regulate LGR4 via the mTORC1/Stat3 signal activation.
文摘动脉性肺动脉高压(Pulmonary Arterial Hypertension, PAH),即第1大类肺动脉高压,是一种以高发病率和死亡率为特点的血管疾病,其主要表现是肺血管重塑和肺血管阻力增加,最终导致右心室衰竭甚至死亡。骨形态发生蛋白9 (Bone Morphogenetic Protein 9, BMP9)属于转化生长因子β (Transforming Growth Factor-β, TGF-β)家族,主要由肝脏星状细胞产生,随血液循环到肺血管内皮细胞上与受体结合,在PAH中发挥相应的生物学效应,但是部分研究结果是相互矛盾的。本文对PAH中BMP9的信号通路及BMP9对肺血管内皮细胞作用的研究和进展进行综述。Arterial pulmonary hypertension (PAH), also known as the group 1 pulmonary hypertension, is a vascular disease characterized by high morbidity and mortality, and its main manifestations are pulmonary vascular remodeling and increased pulmonary vascular resistance, which eventually leads to right ventricular failure and even death. Bone Morphogenetic Protein 9 (BMP9) belongs to the Transforming Growth Factor-β (TGF-β) family, which is mainly produced by hepatic stellate cells, which circulates to pulmonary vascular endothelial cells with blood and bind to their receptors to exert corresponding biological effects in PAH, but some research results are contradictory. This article briefly reviews the BMP9 signaling pathway in PAH and the research and progress of BMP9 on pulmonary vascular endothelial cells.
基金supported by the National Natural Science Foundation of China(81702154)partly from the Chongqing Science and Technology Commission(cstc2020jcyj-cxttX0001)+1 种基金the State Key Laboratory of Silkworm Genome Biology(SKLSGB161718-4)the National Institutes of Health(NIH)Research Project Grant Program(DE030480).
文摘Silk as a natural biomaterial is considered as a promising bone substitute in tissue regeneration.Sericin and fibroin are the main components of silk and display unique features for their programmable mechanical properties,biocompatibility,biodegradability and morphological plasticity.It has been reported that sericin recombinant growth factors(GFs)can support cell proliferation and induce stem cell differentiation through cross-talk of signaling pathways during tissue regeneration.The transgenic technology allows the productions of bioactive heterologous GFs as fusion proteins with sericin,which are then fabricated into solid matrix or hydrogel format.Herein,using an injectable hydrogel derived from transgenic platelet-derived GF(PDGF)-BB silk sericin,we demonstrated that the PDGF-BB sericin hydrogel effectively augmented osteogenesis induced by bone morphogenetic protein(BMP9)-stimulated mesenchymal stem cells(MSCs)in vivo and in vitro,while inhibiting adipogenic differentiation.Further gene expression and protein-protein interactions studies demonstrated that BMP9 and PDGF-BB synergistically induced osteogenic differentiation through the cross-talk between Smad and Stat3 pathways in MSCs.Thus,our results provide a novel strategy to encapsulate osteogenic factors and osteoblastic progenitors in transgenic sericin-based hydrogel for robust bone tissue engineering.
文摘目的:研究骨形态发生蛋白9(Bone morphogenetic protein 9,BMP9)对乳腺癌细胞MDA-MB-231增殖、周期及凋亡的影响。方法:半定量RT-PCR法检测高转移性乳腺癌细胞MDA-MB-231和低转移性乳腺癌细胞MCF-7中BMP9的表达,用人BMP9重组腺病毒(AdBMP9)感染MDA-MB-231细胞作为实验组,含GFP的空载腺病毒(AdGFP)感染MDA-MB-231细胞作为对照组,MTT法观察细胞增殖的变化,流式细胞仪分析周期及凋亡的变化。结果:半定量RT-PCR显示,乳腺癌细胞MDA-MB-231中未检测到BMP9 mRNA的表达;MTT结果显示,AdBMP9处理可引起MDA-MB-231细胞增殖抑制,第5 d MDA-MB-231/BMP9组的细胞增殖率(0.8860±0.0532)显著低于MDA-MB-231/GFP组(1.2240±0.1031)(P<0.05);流式细胞仪分析结果显示,腺病毒感染后第2 d和第3 d,MDA-MB-231/BMP9组细胞周期各相发生变化,G2/M期细胞明显增加,分别为MDA-MB-231/GFP组的3.2倍和2.4倍(P<0.05);同时BMP9可诱发细胞凋亡,腺病毒感染后第3 d MDA-MB-231/BMP9组细胞凋亡百分率(31.55%±8.26%)明显高于MDA-MB-231/GFP组(3.80%±0.46%)(P<0.05)。结论:提示BMP9可通过阻滞细胞周期进程和诱导细胞凋亡来抑制乳腺癌细胞MDA-MB-231的增殖。
基金supported by the National Natural Science Foundation of China,PRC(No.#82002312 and#81972069)This project was also supported by Science and Technology Research Program of Chongqing Education Commission,PRC(No.#KJQN202100431 and#KJZD-M202100401)+1 种基金Candidate of Tip-Top Talent of the First Affiliated Hospital of Chongqing Medical University,PRC(No.BJRC2021-04)Natural Science Foundation of Chongqing Science and Technology Commission,PRC(No.#cstc2018jcyjAX0088).
文摘BMP9 mediated osteogenic differentiation mechanisms of MSCs were widely explored, however, mechanisms of BMP9-induced angiogenesis still need to be clarified. We previously characterized that Notch1 promoted BMP9-induced osteogenesis–angiogenesis coupling process in mesenchymal stem cells (MSCs). Here, we explored the underlying mechanisms of lncRNA H19 (H19) mediated regulation of BMP9-induced angiogenesis through activating Notch1 signaling. We demonstrated that basal expression level of H19 was high in MSCs, and silencing H19 attenuates BMP9-induced osteogenesis and angiogenesis of MSCs both in vitro and in vivo. Meanwhile, we identified that BMP9-induced production of CD31+ cells was indispensable for BMP9-induced bone formation, and silencing H19 dramatically blocked BMP9-induced production of CD31^(+) cells. In addition, we found that down-regulation of H19 inhibited BMP9 mediated blood vessel formation and followed subsequent bone formation in vivo. Mechanistically, we clarified that H19 promoted p53 phosphorylation by direct interacting and phosphorylating binding, and phosphorylated p53 potentiated Notch1 expression and activation of Notch1 targeting genes by binding on the promoter area of Notch1 gene. These findings suggested that H19 regulated BMP9-induced angiogenesis of MSCs by promoting the p53-Notch1 angiogenic signaling axis.
基金We thank Professor T.C.He(Medical Center of University of Chicago,Chicago IL,USA)for his kind provision of the recombinant adenoviruses.
文摘Despite advances in screening and treatment,colon cancer remains one of the leading causes of cancer-related death.Finding novel and useful drug treatment targets is also an urgent need for clinical applications.Tetrandrine(Tet)is extracted from the Chinese medicinal herbal medicine,which is a well-known calcium blocker with a variety of pharmacological activities,including anti-cancer.In this study,we recruited cell viability assay,flow cytometry analysis,cloning formation to confirm that Tet can inhibit the proliferation of SW620 cells,and induce apoptosis.Mechanically,we confirmed that Tet up-regulates the mRNA and protein level of BMP9 in SW620 cells.Over-expression BMP9 enhances the anticancer effects of Tet in SW620 cells,but these effects can be partly reversed by silencing BMP9.Also,Tet reduces phosphorylation of Aktl/2/3 in SW620 cells,which could be elevated by overexpressed BMP9 and impaired by silencing BMP9.Furthermore,we demonstrated that Tet reduces phosphorylated PTEN,which can be promoted by overexpressed BMP9,analogously also be attenuated through silencing BMP9.Finally,we introduced a xenograft tumor model to investigate the anti-proliferative effect of Tet,further to explore the effects of BMP9 and PTEN in SW620 cells.Our findings suggested that the anti-cancer activity of Tet in SW620 cells may be mediated partly by up-regulating BMP9,followed by inactivation PI3K/Akt through up-regulating PTEN at least.
