这项工作已经得到了哥伦布章9671（安大略省温莎市），以及CIHR弗雷德里克班廷和查尔斯·加拿大研究生奖学金颁发给丹尼斯·马骑士。感谢他们的协助，你罗伯特·霍奇和伊丽莎白菲达尔戈达席尔瓦随着时间推移显微镜。谢谢凯蒂Facecchia时间推移显微镜视频编辑。我们也想感谢这个手稿的严格审查Sudipa六月查特吉和Phillip特伦布莱。这项工作是致力于记忆凯文Couvillon谁失去了对抗癌症的斗争，他在2010年。

<ol>
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J. Oncol. 38, 1549-1556 (2011).</li><li>Collins, J., Rinner, U., Moser, M., Hudlicky, T., Ghiviriga, I., Romero, A. E., Kornienko, A., Ma, D., Griffin, C., Pandey, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chemoenzymatic+synthesis+of+Amaryllidaceae+constituents+and+biological+evaluation+of+their+C-1+analogues.+The+next+generation+synthesis+of+7-deoxypancratistatin+and+trans-dihydrolycoricidine.">Chemoenzymatic synthesis of Amaryllidaceae constituents and biological evaluation of their C-1 analogues. The next generation synthesis of 7-deoxypancratistatin and trans-dihydrolycoricidine.</a> J. Org. Chem. 75, 3069-3084 (2010).</li><li>Zhang, G., Gurtu, V., Kain, S. 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U., Haj-Yehia, A., Levi-Schaffer, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Role+of+reactive+oxygen+species+(ROS)+in+apoptosis+induction.">Role of reactive oxygen species (ROS) in apoptosis induction.</a> Apoptosis. 5, 415-418 (2000).</li><li>Batandier, C., Leverve, X., Fontaine, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Opening+of+the+mitochondrial+permeability+transition+pore+induces+reactive+oxygen+species+production+at+the+level+of+the+respiratory+chain+complex+I.">Opening of the mitochondrial permeability transition pore induces reactive oxygen species production at the level of the respiratory chain complex I.</a> J Biol Chem. 279, 17197-17204 (2004).</li><li>Lefranc, F., Sauvage, S., Goietsenoven, G. V. a. n., Mégalizzi, V., Lamoral-Theys, D., Debeir, O., Spiegl-Kreinecker, S., Berger, W., Mathieu, V., Decaestecker, C., Kiss, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Narciclasine,+a+plant+growth+modulator,+activates+Rho+and+stress+fibers+in+glioblastoma+cells.">Narciclasine, a plant growth modulator, activates Rho and stress fibers in glioblastoma cells.</a> Mol. Cancer Ther. 8, 1739-1750 (2009).</li><li>Warburg, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=On+the+origin+of+cancer+cells.">On the origin of cancer cells.</a> Science. 123, 309-314 (1956).</li><li>Heerdt, B. G., Houston, M. A., Augenlicht, L. 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没有利益冲突的声明。

PST和类似的化合物已被证实具有抗癌特性11-15,21。我们以前曾报道自然的PST线粒体选择性破坏肿瘤细胞，从而诱导细胞凋亡12,14凋亡因子的释放。这是最有可能的，JCTH-4的行为，通过相同的机制;造成JCTH-4 TMRM染色（ 图5a），并增加从SH-SY5Y细胞中分离出线粒体ROS生成在MCF7细胞基质金属蛋白酶的崩溃（ 图5B），线粒体功能障碍的指标。因此，诱导细胞凋亡由J...

<table border="1"><tbody><tr><td> 材料名称 </td><td> 公司 </td><td> 目录编号 </td></tr><tr><td> SH-SY5Y细胞线</td><td> ATCC </td><td> CRL-2266 </td></tr><tr><td>杜尔贝科的修改老鹰中等的F-12火腿</td><td> Sigma-Aldrich公司</td><td> 51448C </td></tr><tr><td>胎牛血清</td><td> Gibco BRL公司</td><td> 16000-044 </td></tr><tr><td> MCF7细胞株</td><td> ATCC </td><td> HTB-22 </td></tr><tr><td>的RPMI-1640培养基</td><td> Sigma-Aldrich公司</td><td> ŕ0883 </td></tr><tr><td>显然正常的人类胎儿成纤维细胞株（NFF的） </td><td> coriell研究所医学研究</td><td> AG04431B </td></tr><tr><td>杜尔贝科的改良Eagle培养基高糖</td><td> Thermo Scientific的</td><td> SH30022.01 </td></tr><tr><td>枸橼酸他莫昔芬盐</td><td> Sigma-Aldrich公司</td><td> T9262 </td&gt; </tr><tr><td> 35毫米的玻璃底培养皿</td><td> MatTek </td><td> P35G-014-C的</td></tr><tr><td>徕卡DMI6000 B的反向显微镜</td><td>徕卡公司</td><td> N / A </td></tr><tr><td> Hoechst 33342荧光染料</td><td>分子探针</td><td> H3570 </td></tr><tr><td>徕卡马克内部评级倒置荧光显微镜</td><td>徕卡公司</td><td> N / A </td></tr><tr><td> Annexin V的AlexaFluor-488 </td><td> Invitrogen公司</td><td> A13201 </td></tr><tr><td>台盼蓝溶液</td><td> Sigma-Aldrich公司</td><td> T8154-20ML </td></tr><tr><td>血球</td><td> Fisher Scientific则</td><td> 267110 </td></tr><tr><td> WST-1试剂</td><td>罗氏应用科学部</td><td> 11644807001 </td></tr><tr><td> wallac维克多3 1420多标记计数器</td><td>珀金埃尔默</td><td> 1420-011 </td></tr><tr><td>四甲甲酯（TMRM） </td><td> Gibco BRL公司</td><td>的T-668 </td></tr><tr><td>玻璃组织磨床</td><td> Fisher Scientific则</td><td> K8885300-0002 </td></tr><tr><td> BioRad公司蛋白检测</td><td> BIO-RAD实验室</td><td> 500-0001Bottom的表格</td></tr><tr><td> amplex红</td><td> Invitrogen公司</td><td> A12222 </td></tr><tr><td>辣根过氧化物酶（HRP） </td><td> Sigma-Aldrich公司</td><td> P8125 </td></tr><tr><td> SpectraMax双子座XPS </td><td>分子器件</td><td> 3126666 </td></tr><tr><td>提高兔抗-LC3的抗体</td><td> Novus公司生物制品</td><td> NB100-2220 </td></tr><tr><td>抗小鼠HRP标记的二抗</td><td> abcam </td><td> ab6728 </td></tr><tr><td>抗兔HRP标记的二抗</td><td> abcam </td><td> ab6802 </td></tr><tr><td>化学发光法过氧化物酶底物</td><td> Sigma-Aldrich公司</td><td> CPS160 </td></tr><tr><td> monodansylcadaverine </td><td> Sigma-Aldrich公司</td><td> 30432 </td></tr></tbody></table>

enhancement of apoptotic and autophagic induction by a novel synthetic c-1 analogue of 7-deoxypancratistatin in human breast adenocarcinoma and neuroblastoma cells with tamoxifen

乳腺癌是最常见的癌症之一，除在北美的妇女。目前许多抗癌药物，包括电离辐射，通过DNA损伤诱导细胞凋亡。不幸的是，这种治疗方法有非选择性的肿瘤细胞，在正常细胞产生类似的毒性。我们已经报道（PST），通过天然化合​​物pancratistatin的选择性诱导肿瘤细胞凋亡。最近，一种新的PST模拟，7 deoxypancratistatin（JCTH-4）的C-1乙酰衍生物，从头合成产生，它在多种癌细胞株展品媲美选择性诱导凋亡活性。最近，自噬已被牵连在恶性肿瘤亲生存和死亡机制在化疗反应。三苯氧胺（TAM）总是表现出亲生存在许多癌症细胞自噬的诱导。在这项研究中，JCTH-4单独和谭相结合的疗效，诱导细胞死亡在人类乳腺癌canceR（MCF7细胞）和神经母细胞瘤细胞（SH-SY5Y细胞）进行了评估。谭单独诱导细胞自噬，但单独JCTH-4微不足道的细胞死亡而造成的一些诱导的自噬诱导细胞凋亡的重要。有趣的是，组合的治疗取得了大幅增加在细胞凋亡和自噬的诱导。我们监测的组合使用治疗时间推移显微镜MCF7细胞的形态学变化时间依赖性接受TAM诱导细胞自噬，JCTH-4诱导的细胞凋亡和自噬，并加速细胞死亡。我们已经证明了这些化合物诱导细胞凋亡/自噬在这些癌细胞的线粒体定位。重要的是，这些治疗并不影响非癌的人成纤维细胞的生存。因此，这些结果表明，JCTH-4在与谭相结合，可以作为一个安全和非常强大的抗癌疗法对乳腺癌和神经母细胞瘤细胞。

Watch this Scientific Journal Video about Enhancement of Apoptotic and Autophagic Induction by a Novel Synthetic C-1 Analogue of 7-deoxypancratistatin in Human Breast Adenocarcinoma and Neuroblastoma 

Enhancement of Apoptotic and Autophagic Induction by a Novel Synthetic C-1 Analogue of 7-deoxypancratistatin in Human Breast Adenocarcinoma and Neuroblastoma Cells with Tamoxifen

我们已合成了新型防癌抗癌作为本地pancratistatin活动相媲美的pancratistatin模拟;有趣的是，与他莫昔芬治疗组合的产生在细胞凋亡和自噬的诱导线粒体与非癌成纤维细胞的影响最小的目标大幅提高。因此，与他莫昔芬结合JCTH-4可以提供一个安全的抗癌疗法。

增强细胞凋亡和自噬的诱导合成一种新型的C-1 7-deoxypancratistatin的类似物在人类乳癌细胞和神经母细胞瘤与他莫昔芬

Breast cancer is one of the most common cancers amongst women in North America. Many current anti-cancer treatments, including ionizing radiation, induce ...

enhancement-apoptotic-autophagic-induction-novel-synthetic-c-1

Research

JoVE Journal

Medicine

18.6K 次观看.  University of Windsor. 我们已合成了新型防癌抗癌作为本地pancratistatin活动相媲美的pancratistatin模拟;有趣的是，与他莫昔芬治疗组合的产生在细胞凋亡和自噬的诱导线粒体与非癌成纤维细胞的影响最小的目标大幅提高。因此，与他莫昔芬结合JCTH-4可以提供一个安全的抗癌疗法。

视频: Enhancement of Apoptotic and Autophagic Induction by a Novel Synthetic C-1 Analogue of 7-deoxypancratistatin in Human Breast Adenocarcinoma and Neuroblastoma Cells with Tamoxifen

Isolation, Characterization and Comparative Differentiation of Human Dental Pulp Stem Cells Derived from Permanent Teeth by Using Two Different Methods

Developing wisdom teeth are easy-accessible source of stem cells during the adulthood which could be obtained by routine orthodontic treatments. Human pulp-derived stem cells (hDPSCs) possess high proliferation potential with multi-lineage differentiation capacity compare to the ordinary source of adult stem cells1-8; therefore, hDPSCs could be the good candidates for autologous transplantation in tissue engineering and regenerative medicine. Along with these benefits, possessing the mesenchymal stem cells (MSC) features, such as immunolodulatory effect, make hDPSCs more valuable, even in the case of allograft transplantation6,9,10. Therefore, the primary step for using this source of stem cells is to select the best protocol for isolating hDPSCs from pulp tissue. In order to achieve this goal, it is crucial to investigate the effect of various isolation conditions on different cellular behaviors, such as their common surface markers &amp; also their differentiation capacity.
Thus, here we separate human pulp tissue from impacted third molar teeth, and then used both existing protocols based on literature, for isolating hDPSCs,11-13 i.e. enzymatic dissociation of pulp tissue (DPSC-ED) or outgrowth from tissue explants (DPSC-OG). In this regards, we tried to facilitate the isolation methods by using dental diamond disk. Then, these cells characterized in terms of stromal-associated Markers (CD73, CD90, CD105 &amp; CD44), hematopoietic/endothelial Markers (CD34, CD45 &amp; CD11b), perivascular marker, like CD146 and also STRO-1. Afterwards, these two protocols were compared based on the differentiation potency into odontoblasts by both quantitative polymerase chain reaction (QPCR) &amp; Alizarin Red Staining. QPCR were used for the assessment of the expression of the mineralization-related genes (alkaline phosphatase; ALP, matrix extracellular phosphoglycoprotein; MEPE &amp; dentin sialophosphoprotein; DSPP).14

The method described isolation and characterization of human Dental Pulp Stem Cells (hDPSCs) by using either enzymatic dissociation of pulp (DPSC-ED) or direct outgrowth of stem cells from pulp tissue explants (DPSC-OG). Then followed by in vitro comparative differentiation of both types of hDPSCs into odontoblasts.

人牙髓干细胞的分离，鉴定和比较性恒牙使用两种不同的方法，得出

Developing wisdom teeth are easy-accessible source of stem cells during the adulthood which could be obtained by routine orthodontic treatments. Human ...

科研

医学

Cell Death Associated with Abnormal Mitosis Observed by Confocal Imaging in Live Cancer Cells

Phenanthrene derivatives acting as potent PARP1 inhibitors prevented the bi-focal clustering of supernumerary centrosomes in multi-centrosomal human cancer cells in mitosis. The phenanthridine PJ-34 was the most potent molecule. Declustering of extra-centrosomes causes mitotic failure and cell death in multi-centrosomal cells. Most solid human cancers have high occurrence of extra-centrosomes. The activity of PJ-34 was documented in real-time by confocal imaging of live human breast cancer MDA-MB-231 cells transfected with vectors encoding for fluorescent &gamma;-tubulin, which is highly abundant in the centrosomes and for fluorescent histone H2b present in the chromosomes. Aberrant chromosomes arrangements and de-clustered &gamma;-tubulin foci representing declustered centrosomes were detected in the transfected MDA-MB-231 cells after treatment with PJ-34. Un-clustered extra-centrosomes in the two spindle poles preceded their cell death. These results linked for the first time the recently detected exclusive cytotoxic activity of PJ-34 in human cancer cells with extra-centrosomes de-clustering in mitosis, and mitotic failure leading to cell death. According to previous findings observed by confocal imaging of fixed cells, PJ-34 exclusively eradicated cancer cells with multi-centrosomes without impairing normal cells undergoing mitosis with two centrosomes and bi-focal spindles. This cytotoxic activity of PJ-34 was not shared by other potent PARP1 inhibitors, and was observed in PARP1 deficient MEF harboring extracentrosomes, suggesting its independency of PARP1 inhibition. Live confocal imaging offered a useful tool for identifying new molecules eradicating cells during mitosis.

The cytotoxic activity of the phenanthridine PJ-34 in cancer cells undergoing mitosis was documented in real time by live confocal imaging. PJ-34 eradicated human breast cancer MDA-MB-231 cells harboring extra-centrosomes in mitosis. Unlike normal bi-focal mitosis, the extra-centrosomes were not clustered in the two spindle poles in the presence of PJ-34.

共聚焦成像在肝癌细胞中观察到的异常有丝分裂细胞死亡相关

Phenanthrene derivatives acting as potent PARP1 inhibitors prevented the bi-focal clustering of supernumerary centrosomes in multi-centrosomal human ...

Initiation of Metastatic Breast Carcinoma by Targeting of the Ductal Epithelium with Adenovirus-Cre: A Novel Transgenic Mouse Model of Breast Cancer

Breast cancer is a heterogeneous disease involving complex cellular interactions between the developing tumor and immune system, eventually resulting in exponential tumor growth and metastasis to distal tissues and the collapse of anti-tumor immunity. Many useful animal models exist to study breast cancer, but none completely recapitulate the disease progression that occurs in humans. In order to gain a better understanding of the cellular interactions that result in the formation of latent metastasis and decreased survival, we have generated an inducible transgenic mouse model of YFP-expressing ductal carcinoma that develops after sexual maturity in immune-competent mice and is driven by consistent, endocrine-independent oncogene expression. Activation of YFP, ablation of p53, and expression of an oncogenic form of K-ras was achieved by the delivery of an adenovirus expressing Cre-recombinase into the mammary duct of sexually mature, virgin female mice. Tumors begin to appear 6 weeks after the initiation of oncogenic events. After tumors become apparent, they progress slowly for approximately two weeks before they begin to grow exponentially. After 7-8 weeks post-adenovirus injection, vasculature is observed connecting the tumor mass to distal lymph nodes, with eventual lymphovascular invasion of YFP+ tumor cells to the distal axillary lymph nodes. Infiltrating leukocyte populations are similar to those found in human breast carcinomas, including the presence of &#945;&#946; and &#947;&#948; T cells, macrophages and MDSCs. This unique model will facilitate the study of cellular and immunological mechanisms involved in latent metastasis and dormancy in addition to being useful for designing novel immunotherapeutic interventions to treat invasive breast cancer.

Activation of latent mutations with adenovirus-Cre into the mammary ductal system results in a clinically relevant metastatic breast cancer. Incorporation of a YFP promoter allows tracking of distal metastatic tumor cells. This model is useful to study latent metastasis, anti-tumor immunity, and for designing novel immunotherapies to treat breast cancer.

乳腺癌的一种新的转基因小鼠模型：转移性乳腺癌通过瞄准导管上皮与腺病毒Cre重组酶的启动

Breast cancer is a heterogeneous disease involving complex cellular interactions between the developing tumor and immune system, eventually resulting in ...

Profiling of Estrogen-regulated MicroRNAs in Breast Cancer Cells

Estrogen plays vital roles in mammary gland development and breast cancer progression. It mediates its function by binding to and activating the estrogen receptors (ERs), ER&#945;, and ER&#946;. ER&#945; is frequently upregulated in breast cancer and drives the proliferation of breast cancer cells. The ERs function as transcription factors and regulate gene expression. Whereas ER&#945;&#39;s regulation of protein-coding genes is well established, its regulation of noncoding microRNA (miRNA) is less explored. miRNAs play a major role in the post-transcriptional regulation of genes, inhibiting their translation or degrading their mRNA. miRNAs can function as oncogenes or tumor suppressors&#160;and are also promising biomarkers. Among the miRNA assays available, microarray and quantitative real-time polymerase chain reaction (qPCR) have been extensively used to detect and quantify miRNA levels. To identify miRNAs regulated by estrogen signaling in breast cancer, their expression in ER&#945;-positive breast cancer cell lines were compared before and after estrogen-activation using both the &#181;Paraflo-microfluidic microarrays and Dual Labeled Probes-low density arrays. Results were validated using specific qPCR assays, applying both Cyanine dye-based and Dual Labeled Probes-based chemistry. Furthermore, a time-point assay was used to identify regulations over time. Advantages of the miRNA assay approach used in this study is that it enables a fast screening of mature miRNA regulations in numerous samples, even with limited sample amounts. The layout, including the specific conditions for cell culture and estrogen treatment, biological and technical replicates, and large-scale screening followed by in-depth confirmations using separate techniques, ensures a robust detection of miRNA regulations,&#160;and eliminates false positives and other artifacts. However, mutated or unknown miRNAs, or regulations at the primary and precursor transcript level, will not be detected. The method presented here represents a thorough investigation of estrogen-mediated miRNA regulation.

Molecular signaling through both estrogen and microRNAs are critical in breast cancer development and growth. Estrogen activates the estrogen receptors, which are transcription factors. Many transcription factors can regulate the expression of microRNAs, and estrogen-regulated microRNAs can be profiled using different large-scale techniques.

在乳腺癌细胞雌激素调节小分子RNA的剖析

Estrogen plays vital roles in mammary gland development and breast cancer progression. It mediates its function by binding to and activating the estrogen ...

Utilization of the Soft Agar Colony Formation Assay to Identify Inhibitors of Tumorigenicity in Breast Cancer Cells

Given the inherent difficulties in investigating the mechanisms of tumor progression in vivo, cell-based assays such as the soft agar colony formation assay (hereafter called soft agar assay), which measures the ability of cells to proliferate in semi-solid matrices, remain a hallmark of cancer research. A key advantage of this technique over conventional 2D monolayer or 3D spheroid cell culture assays is the close mimicry of the 3D cellular environment to that seen in vivo. Importantly, the soft agar assay also provides an ideal tool to rigorously test the effects of novel compounds or treatment conditions on cell proliferation and migration. Additionally, this assay enables the quantitative assessment of cell transformation potential within the context of genetic perturbations. We recently identified peptidylarginine deiminase 2 (PADI2) as a potential breast cancer biomarker and therapeutic target. Here we highlight the utility of the soft agar assay for preclinical anti-cancer studies by testing the effects of the PADI inhibitor, BB-Cl-amidine (BB-CLA), on the tumorigenicity of human ductal carcinoma in situ (MCF10DCIS) cells.

Here, we document the use of the soft agar colony formation assay to test the effects of a peptidylarginine deiminase (PADI) enzyme inhibitor, BB-Cl-amidine, on breast cancer tumorigenicity in vitro.

软琼脂集落形成实验利用识别的致瘤性乳腺癌细胞抑制剂

Given the inherent difficulties in investigating the mechanisms of tumor progression in vivo, cell-based assays such as the soft agar colony formation ...

Studying Pancreatic Cancer Stem Cell Characteristics for Developing New Treatment Strategies

Pancreatic ductal adenocarcinoma (PDAC) contains a subset of exclusively tumorigenic cancer stem cells (CSCs) which have been shown to drive tumor initiation, metastasis and resistance to radio- and chemotherapy. Here we describe a specific methodology for culturing primary human pancreatic CSCs as tumor spheres in anchorage-independent conditions. Cells are grown in serum-free, non-adherent conditions in order to enrich for CSCs while their more differentiated progenies do not survive and proliferate during the initial phase following seeding of single cells. This assay can be used to estimate the percentage of CSCs present in a population of tumor cells. Both size (which can range from 35 to 250 micrometers) and number of tumor spheres formed represents CSC activity harbored in either bulk populations of cultured cancer cells or freshly harvested and digested tumors 1,2. Using this assay, we recently found that metformin selectively ablates pancreatic CSCs; a finding that was subsequently further corroborated by demonstrating diminished expression of pluripotency-associated genes/surface markers and reduced in vivo tumorigenicity of metformin-treated cells. As the final step for preclinical development we treated mice bearing established tumors with metformin and found significantly prolonged survival. Clinical studies testing the use of metformin in patients with PDAC are currently underway (e.g., NCT01210911, NCT01167738, and NCT01488552). Mechanistically, we found that metformin induces a fatal energy crisis in CSCs by enhancing reactive oxygen species (ROS) production and reducing mitochondrial transmembrane potential. In contrast, non-CSCs were not eliminated by metformin treatment, but rather underwent reversible cell cycle arrest. Therefore, our study serves as a successful example for the potential of in vitro sphere formation as a screening tool to identify compounds that potentially target CSCs, but this technique will require further in vitro and in vivo validation to eliminate false discoveries.

Pancreatic cancer stem cells (CSCs) can be expanded in vitro using the anchorage-independent sphere culture technique, which represents a powerful tool to study CSC biology and can serve as the first step to develop novel CSC-targeting therapies. Here the methodology for expanding, analyzing and targeting of pancreatic CSCs is provided.

研究胰腺癌干细胞特性开发新的治疗策略

Pancreatic ductal adenocarcinoma (PDAC) contains a subset of exclusively tumorigenic cancer stem cells (CSCs) which have been shown to drive tumor ...

Systems Biology of Metabolic Regulation by Estrogen Receptor Signaling in Breast Cancer

With the advent of the -omics approaches our understanding of the chronic diseases like cancer and metabolic syndrome has improved. However, effective mining of the information in the large-scale datasets that are obtained from gene expression microarrays, deep sequencing experiments or metabolic profiling is essential to uncover and then effectively target the critical regulators of diseased cell phenotypes. Estrogen Receptor &#945; (ER&#945;) is one of the master transcription factors regulating the gene programs that are important for estrogen responsive breast cancers. In order to understand to role of ER&#945; signaling in breast cancer metabolism we utilized transcriptomic, cistromic and metabolomic data from MCF-7 cells treated with estradiol. In this report we described generation of samples for RNA-Seq, ChIP-Seq and metabolomics experiments and the integrative computational analysis of the obtained data. This approach is useful in delineating novel molecular mechanisms and gene regulatory circuits that are regulated by a particular transcription factor which impacts metabolism of normal or diseased cells.

Integration of data from genome-wide sequencing experiments and metabolomics experiments is a challenge. In this paper we report, for the first time, generation, analysis and integration of transcriptome, cistrome and metabolome data from breast cancer cells treated with estradiol.

乳腺癌代谢调控的系统生物学雌激素受体信号

With the advent of the -omics approaches our understanding of the chronic diseases like cancer and metabolic syndrome has improved. However, effective ...

Induction of Accelerated Atherosclerosis in Mice: The "Wire-Injury" Model

Atherosclerosis is a proliferative fibro-inflammatory disease developing in the arterial wall, inducing a deficient blood flow or a lack of blood flow. Moreover, by rupture of the defective vascular wall, atherosclerosis induces occlusive thrombus formation, which represents the main cause of myocardial infarction or stroke and the most frequent cause of death. Despite the advances in the cardiovascular field, many questions remain unanswered, and additional basic research is essential to improve our understanding of the molecular mechanisms during atherosclerosis and its effects. Due to limited clinical studies, there is a need for representative animal models recreating atherosclerotic conditions such as neointima formation after stent implantation, balloon angioplasty, or endarterectomy. Since the mouse presents many advantages and is the most frequently used model for studying molecular processes, the current study proposes an invasive procedure of endothelial denudation, also known as the wire-injury model, which is representative of the human condition of neointima formation in arteries after revascularization procedures.

Induction of Accelerated Atherosclerosis in Mice: The &quot;Wire-Injury&quot; Model

This study describes an invasive procedure for the induction of accelerated atherosclerosis in mice. In comparison to other methods using electric- or cryo-induced injury, mechanical-induced injury mimics the human condition of restenosis after revascularization therapies and is ideal for the study of the molecular mechanisms involved.

小鼠加速动脉硬化的诱导："电线损伤"模型

Atherosclerosis is a proliferative fibro-inflammatory disease developing in the arterial wall, inducing a deficient blood flow or a lack of blood flow. ...

A Novel Clinical Grade Isolation Method for Human Kidney Perivascular Stromal Cells

Mesenchymal Stromal Cells (MSCs) are tissue homeostatic and immune modulatory cells that have shown beneficial effects in kidney diseases and transplantation. Perivascular Stromal Cells (PSCs) share characteristics with bone marrow MSCs (bmMSCs). However, they also possess, most likely due to local imprinting, tissue-specific properties and play a role in local tissue homeostasis. This tissue specificity may result in tissue specific repair, also within the human kidney. We previously showed that human kidney PSCs (kPSCs) have enhanced kidney epithelial wound healing whereas bmMSCs did not have this potential. Moreover, kPSCs can ameliorate kidney injury in vivo. Therefore, kPSCs constitute an interesting source for cell therapy, particularly for kidney diseases and renal transplantation. Here we show the detailed isolation and culture method for kPSCs from transplant-grade human kidneys based on whole-organ perfusion of digestive enzymes via the renal artery and enrichment for the perivascular marker NG2. In this way, large cell quantities can be obtained that are suitable for cellular therapy.

Here we present a novel clinical grade isolation and culture method for kidney Perivascular Stromal Cells (kPSCs) based on whole organ perfusion with digestive enzymes and NG2-cell enrichment. With this method, it is possible to acquire sufficient cell numbers for cellular therapy.

人类肾脏血管周围间质细胞新型临床分级分离方法

Mesenchymal Stromal Cells (MSCs) are tissue homeostatic and immune modulatory cells that have shown beneficial effects in kidney diseases and ...

Characterization of Immune Cells in Human Adipose Tissue by Using Flow Cytometry

Infiltration of immune cells in the subcutaneous and visceral adipose tissue (AT) deposits leads to a low-grade inflammation contributing to the development of obesity-associated complications such as type 2 diabetes. To quantitatively and qualitatively investigate the immune cell subsets in human AT deposits, we have developed a flow cytometry approach. The stromal vascular fraction (SVF), containing the immune cells, is isolated from subcutaneous and visceral AT biopsies by collagenase digestion. Adipocytes are removed after centrifugation. The SVF cells are stained for multiple membrane-bound markers selected to differentiate between immune cell subsets and analyzed using flow cytometry. As a result of this approach, pro- and anti-inflammatory macrophage subsets, dendritic cells (DCs), B-cells, CD4+ and CD8+ T-cells, and NK cells can be detected and quantified. This method gives detailed information about immune cells in AT and the amount of each specific subset. Since there are numerous fluorescent antibodies available, our flow cytometry approach can be adjusted to measure various other cellular and intracellular markers of interest.

This article describes a method to analyze immune cell content of adipose tissue by isolation of immune cells from adipose tissue and subsequent analysis using flow cytometry.