Promotion of proliferation of luminal B breast cancer cells by mesenchymal stem cells and its underlying molecular mechanisms

目的分析人脐带间充质干细胞(hUC-MSCs)对Luminal B型乳腺癌细胞生长增殖的影响,并初步探讨其可能的分子机理。方法绿色荧光蛋白(GFP)和荧光素酶共表达慢病毒感染人Luminal B型乳腺癌细胞BT474,并经嘌呤霉素筛选两周后,于荧光显微镜下观察GFP的表达情况,IVIS Kinetic成像系统拍照以观察和记录慢病毒感染后BT474细胞荧光素酶的表达情况;荧光显微镜下直接观察,结合MTS实验分析hUC-MSCs共培养或其浓缩上清处理对GFP和荧光素酶共表达BT474细胞生长增殖的影响;Western blot法检测hUC-MSCs浓缩上清处理对BT474细胞Akt和MAPK信号通路激活情况以及下游细胞周期调控蛋白Cyclin D1表达的影响;常规RT-PCR法检测hUC-MSCs中NRG-1、NRG-2、IGF-Ⅰ、IGF-Ⅱ和EGF等配体的表达。荧光素酶表达强度与细胞数量的相关性经由Excel软件行统计学分析,MTS实验数据则经由SPSS13.0统计软件行统计学分析。结果荧光显微镜和IVIS Kinetic成像系统的观察结果分别证实,GFP和荧光素酶经慢病毒载体系统的介导可在BT474细胞中成功地共表达,且荧光素酶的表达强度与细胞数量呈直线相关。MSCs共培养或其浓缩上清处理均可显著促进Luminal B型乳腺癌细胞BT474的生长增殖,其细胞存活比例分别为各自对照组的148.06%(P<0.005)和147.99%(P<0.001);MSCs浓缩上清处理同时激活BT474细胞内Akt和MAPK信号通路,并上调细胞周期调控蛋白Cyclin D1表达。此外,RT-PCR结果显示,hUC-MSCs中NRG-1和EGF的mRNAs水平呈高表达,而NRG-2、IGF-Ⅰ和IGF-Ⅱ等配体的mRNAs表达也可见。结论 MSCs可通过表达并分泌NRG-1等配体,从而激活Luminal B型乳腺癌细胞BT474的下游Akt和MAPK信号转导通路以上调细胞周期调控蛋白Cyclin D1的表达,进而促进其生长增殖。Objective To investigate the effect of human umbilical cord mesenchymal stem cells(hUC-MSCs) on proliferation of luminal B breast cancer cells and its underlying molecular mechanisms. Methods Human luminal B breast cancer cells BT474 were infected with GFP and luciferase co-expressing lentiviruses and then subjected for selection with Puromycin for 2 weeks. The expression of GFP and luciferase was detected by fluorescent microscopy and IVIS Kinetic image system, respectively. The effect of coculture or treatment with conditioned medium of hUCMSCs on proliferation of BT474 was analyzed with MTS assay. Western blot was carried out to detect the effect of treatment with conditioned medium of hUC-MSCs on the activation of both Akt and MAPK signalings in BT474, as well as the expression of downstream cell cycle regulator Cyclin D1. Regular RT-PCR was applied to analyze the mRNAs expression of ligands such as NRG-1, NRG-2, IGF-Ⅰ, IGF-Ⅱ, and EGF in hUC-MSCs. The correlation between relative luciferase activity and cell number was analyzed with Excel software, while MTS assay data was statistically analyzed with SPSS 13.0 software. Results The co-expression of GFP and luciferase in BT474 via lentiviral expression system was visualized by fluorescent microscopy and IVIS Kinetic image system. The linear correlation between relative luciferase activity and cell number was determined by curve fitting analysis. Coculture or treatment with conditioned medium of hUC-MSC significantly promoted the proliferation of BT474, with survival rates being 148.06 %(P < 0.005)and 147.99 %(P < 0.001)of control, respectively. In addition, treatment with conditioned medium of hUC-MSC was shown to induce activation of both Akt and MAPK signalings, which further upregulated the expression of Cyclin D1. Moreover, high mRNAs expression levels of both NRG-1 and EGF, as well as moderate mRNAs expression levels NRG-2, IGF-Ⅰ, and IGF-Ⅱ were showed by RT-PCR. Conclusion Our results here demonstrated that MSCs may promote the proliferation of luminal B breast cancer cells through paracrine of ligands such as NRG-1, which in turn results in the activation of both Akt and MAPK signalings and upregulation of the expression of Cyclin D1.国家自然科学基金面上项目(81272922);; 福建省自然科学基金面上项目(2016J01577);; 福州总医院院内课题国际合作研究专项(2015G01

施肥对新疆引进欧李生长及营养品质的影响

【目的】施肥对欧李生长及营养品质的影响。【方法】对野生欧李和山西选育的三个欧李品种的基肥、追肥、微肥的根施和叶面喷施处理以及果实营养成分分析。【结果】在肥力水平较高的试验地栽培,定植时不宜施基肥;叶面喷施氯化钙,可以提高野生欧李果实品质;根施氮、磷、钾肥可提高次年坐果率;从野生欧李中筛选出几株优良单株,经过果实品尝和营养成分分析,野生欧李的果实品质比选育的欧李品种好。【结论】欧李的根系发达,抗逆性强,果实营养价值很高,新疆引进欧李种植对干旱地区农业可持续发展有积极作用,具有广阔的应用发展前景

中国空间生命科学40年回顾与展望

我国空间生命科学的探索起源于20世纪60年代,1981年随着空间生命专业委员会的正式成立,依托此专业的学术交流平台,空间生命科学进入多学科并进多机构建设的新阶段.随着中国载人航天及空间探索研究的深入发展,以分支学科或重大问题为牵引,我国在空间生命科学的几个重要领域取得了一系列关键成果.本文从发展历程、研究成果、平台模型、重大项目与后续展望等方面综述了我国空间生命科学40年的发展历程与标志性成果,为后续发展提供借鉴与参考

~(265)Bh(Z=107)同位素的首次观测

在兰州的重离子加速器上用 2 6Mg离子束轰击 2 43 Am靶 ,产生了新同位素 2 65Bh .通过观测新同位素 2 65Bh和它的已知子核 2 61Db和 2 57Lr之间的α衰变的关联 ,实现了对新核素的鉴别 .实验中使用了一套新建立的具有数个探测器对的转轮收集探测系统 .将该系统用于特殊的母 -子核搜索模式 ,从而大大减少了本底 .共测得了 8个 2 65Bh的α衰变关联事件 ;同时 4个已知核 2 64Bh的衰变关联事件也被鉴别出来 .实验测得 2 65Bh的α衰变能量为 (9.2 4± 0 .0 5 )MeV ,半衰期为 0 .94 + 0 .70-0 .3 1s

新同位素~(265)Bh(Z=107)的合成证据

利用兰州重离子加速器提供的26Mg离子束轰击243Am靶, 产生了新同位素265Bh. 实验中用氦喷技术对产物进行传输, 并用一套具有数对探测器组的转轮收集探测系统对产物进行收集和测量. 通过观测265Bh与它的衰变子核261Db及257Lr之间的α衰变的关联, 实现了对新核素的鉴别. 实验测得265Bh的α衰变能量为(9. 24±0. 05)MeV, 半衰期为 0. 94+0. 70 0. 31 s

利用α关联衰变链鉴别新同位素_(107)~(265)Bh

主要介绍了合成和鉴别107号元素的新同位素265Bh的实验装置、实验方法以及实验结果.目标核265Bh是由能量为135MeV的26Mg离子轰击243Am靶,通过融合蒸发反应而产生.反应产物首先由He jet系统传输到装有数个探测器对的转轮收集测量系统,然后依靠母子核遗传关系通过观察新同位素和它们已知子核261Db和257Lr之间的α衰变的关联,来实现对新核素的鉴别.实验测得265Bh的α衰变能量为(9.24±0.05)MeV, 半衰期为0.94+0.70-0.31s.从该实验得出的265Bh的α衰变能量和半寿命能够与理论预言一致