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。

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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

Synthesis and Characterization of an Aspirin-fumarate Prodrug that Inhibits NF&#954;B Activity and Breast Cancer Stem Cells

Inflammation is a cancer hallmark that underlies cancer incidence and promotion, and eventually progression to metastasis. Therefore, adding an anti-inflammatory drug to standard cancer regiments may improve patient outcome. One such drug, aspirin (acetylsalicylic acid, ASA), has been explored for cancer chemoprevention and anti-tumor activity. Besides inhibiting the cyclooxygenase 2-prostaglandin axis, ASA&#39;s anti-cancer activities have also been attributed to nuclear factor &#312;B (NF&#312;B) inhibition. Because prolonged ASA use may cause gastrointestinal toxicity, a prodrug strategy has been implemented successfully. In this prodrug design the carboxylic acid of ASA is masked and additional pharmacophores are incorporated.
This protocol describes how we synthesized an aspirin-fumarate prodrug, GTCpFE, and characterized its inhibition of the NF&#312;B pathway in breast cancer cells and attenuation of the cancer stem-like properties, an important NF&#312;B-dependent phenotype. GTCpFE effectively inhibits the NF&#312;B pathway in breast cancer cell lines whereas ASA lacks any inhibitory activity, indicating that adding fumarate to ASA structure significantly contributes to its activity. In addition, GTCpFE shows significant anti-cancer stem cell activity by blocking mammosphere formation and attenuating the cancer stem cell associated CD44+CD24- immunophenotype. These results establish a viable strategy to develop improved anti-inflammatory drugs for chemoprevention and cancer therapy.

This procedure will demonstrate how we synthesized and characterized the anti-NF&#954;B and anti-cancer stem cell activity of an aspirin-fumarate prodrug.

合成与阿司匹林富马酸酯前药的表征抑制NFκB活动和乳腺癌干细胞

Inflammation is a cancer hallmark that underlies cancer incidence and promotion, and eventually progression to metastasis. Therefore, adding an ...

科研

癌症研究

Therapeutic Gene Delivery and Transfection in Human Pancreatic Cancer Cells using Epidermal Growth Factor Receptor-targeted Gelatin Nanoparticles

More than 32,000 patients are diagnosed with pancreatic cancer in the United States per year and the disease is associated with very high mortality 1. Urgent need exists to develop novel clinically-translatable therapeutic strategies that can improve on the dismal survival statistics of pancreatic cancer patients. Although gene therapy in cancer has shown a tremendous promise, the major challenge is in the development of safe and effective delivery system, which can lead to sustained transgene expression. 
Gelatin is one of the most versatile natural biopolymer, widely used in food and pharmaceutical products. Previous studies from our laboratory have shown that type B gelatin could physical encapsulate DNA, which preserved the supercoiled structure of the plasmid and improved transfection efficiency upon intracellular delivery. By thiolation of gelatin, the sulfhydryl groups could be introduced into the polymer and would form disulfide bond within nanoparticles, which stabilizes the whole complex and once disulfide bond is broken due to the presence of glutathione in cytosol, payload would be released 2-5. Poly(ethylene glycol) (PEG)-modified GENS, when administered into the systemic circulation, provides long-circulation times and preferentially targets to the tumor mass due to the hyper-permeability of the neovasculature by the enhanced permeability and retention effect 6. Studies have shown over-expression of the epidermal growth factor receptor (EGFR) on Panc-1 human pancreatic adenocarcinoma cells 7. In order to actively target pancreatic cancer cell line, EGFR specific peptide was conjugated on the particle surface through a PEG spacer.8
Most anti-tumor gene therapies are focused on administration of the tumor suppressor genes, such as wild-type p53 (wt-p53), to restore the pro-apoptotic function in the cells 9. The p53 mechanism functions as a critical signaling pathway in cell growth, which regulates apoptosis, cell cycle arrest, metabolism and other processes 10. In pancreatic cancer, most cells have mutations in p53 protein, causing the loss of apoptotic activity. With the introduction of wt-p53, the apoptosis could be repaired and further triggers cell death in cancer cells 11. 
Based on the above rationale, we have designed EGFR targeting peptide-modified thiolated gelatin nanoparticles for wt-p53 gene delivery and evaluated delivery efficiency and transfection in Panc-1 cells.

Type B gelatin-based engineered nanovectors system (GENS) was developed for systemic gene delivery and transfection in the treatment of pancreatic cancer. By modification with epidermal growth factor receptor (EGFR) specific peptide on the surface of nanparticles, they could target on EGFR receptor and release plasmid under reducing environment, such as high intracellular glutathione concentrations.

交付使用表皮生长因子在人胰腺癌细胞转染基因受体有针对性的明胶基于工程Nanovectors

More than 32,000 patients are diagnosed with pancreatic cancer in the United States per year and the disease is associated with very high mortality 1. ...

生物工程

Flow Cytometric Detection of Newly-formed Breast Cancer Stem Cell-like Cells After Apoptosis Reversal

Cancer recurrence has long been studied by oncologists while the underlying mechanisms remain unclear. Recently, we and others found that a phenomenon named apoptosis reversal leads to increased tumorigenicity in various cell models under different stimuli. Previous studies have been focused on tracking this process in vitro and in vivo; however, the isolation of real reversed cells has yet to be achieved, which limits our understanding on the consequences of apoptosis reversal. Here, we take advantage of a Caspase-3/7 Green Detection dye to label cells with activated caspases after apoptotic induction. Cells with positive signals are further sorted out by fluorescence-activated cell sorting (FACS) for recovery. Morphological examination under confocal microscopy helps confirm the apoptotic status before FACS. An increase in tumorigenicity can often be attributed to the elevation in the percentage of cancer stem cell (CSC)-like cells. Also, given the heterogeneity of breast cancer, identifying the origin of these CSC-like cells would be critical to cancer treatment. Thus, we prepare breast non-stem cancer cells before triggering apoptosis, isolating caspase-activated cells and performing the apoptosis reversal procedure. Flow cytometry analysis reveals that breast CSC-like cells re-appear in the reversed group, indicating breast CSC-like cells are transited from breast non-stem cancer cells during apoptosis reversal. In summary, this protocol includes the isolation of apoptotic breast cancer cells and detection of changes in CSC percentage in reversed cells by flow cytometry.

Here, we present a protocol to isolate apoptotic breast cancer cells by fluorescence-activated cell sorting and further detect the transition of breast non-stem cancer cells to breast cancer stem cell-like cells after apoptosis reversal by flow cytometry.

细胞凋亡逆转后新形成乳腺癌干细胞样细胞的流式细胞仪检测

Cancer recurrence has long been studied by oncologists while the underlying mechanisms remain unclear. Recently, we and others found that a phenomenon ...

Immunohistochemical Staining of B7-H1 (PD-L1) on Paraffin-embedded Slides of Pancreatic Adenocarcinoma Tissue

B7-H1/PD-L1, a member of the B7 family of immune-regulatory cell-surface proteins, plays an important role in the negative regulation of cell-mediated immune responses through its interaction with its receptor, programmed death-1 (PD-1) 1,2. Overexpression of B7-H1 by tumor cells has been noted in a number of human cancers, including melanoma, glioblastoma, and carcinomas of the lung, breast, colon, ovary, and renal cells, and has been shown to impair anti-tumor T-cell immunity3-8.
Recently, B7-H1 expression by pancreatic adenocarcinoma tissues has been identified as a potential prognostic marker9,10. Additionally, blockade of B7-H1 in a mouse model of pancreatic cancer has been shown to produce an anti-tumor response11. These data suggest the importance of B7-H1 as a potential therapeutic target. Anti-B7-H1 blockade antibodies are therefore being tested in clinical trials for multiple human solid tumors including melanoma and cancers of lung, colon, kidney, stomach and pancreas12.
In order to eventually be able to identify the patients who will benefit from B7-H1 targeting therapies, it is critical to investigate the correlation between expression and localization of B7-H1 and patient response to treatment with B7-H1 blockade antibodies. Examining the expression of B7-H1 in human pancreatic adenocarcinoma tissues through immunohistochemistry will give a better understanding of how this co-inhibitory signaling molecule contributes to the suppression of antitumor immunity in the tumor's microenvironment. The anti-B7-H1 monoclonal antibody (clone 5H1) developed by Chen and coworkers has been shown to produce reliable staining results in cryosections of multiple types of human neoplastic tissues4,8, but staining on paraffin-embedded slides had been a challenge until recently13-18. We have developed the B7-H1 staining protocol for paraffin-embedded slides of pancreatic adenocarcinoma tissues. The B7-H1 staining protocol described here produces consistent membranous and cytoplasmic staining of B7-H1 with little background.

Immunohistochemical Staining of B7-H1 PD-L1 on Paraffin-embedded Slides of Pancreatic Adenocarcinoma Tissue

B7-H1 (PD-L1) and its binding to PD-1 provide a major tumor-induced immunosuppressive signal in the tumor&#x2019;s microenvironment. An immunohistochemical staining technique to characterize the expression and localization of B7-H1 in pancreatic adenocarcinoma is described here.

B7-H1（PD-L1）免疫组化染色石蜡的嵌入式幻灯片的胰腺癌组织

B7-H1/PD-L1, a member  of the B7 family of immune-regulatory cell-surface proteins, plays an important  role in the negative regulation of cell-mediated ...

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 ...

FAM83A 作为宫颈癌靶基因的验证

Knockdown of FAM83A to Verify Its Role in Cervical Cancer Cell Growth and Cisplatin Sensitivity

The exploration of tumor target genes holds paramount importance for the prevention and treatment of cervical cancer. In this study, we outline the steps involved in the identification of a tumor target gene FAM83A in cervical cancer. First, the Cancer Genome Atlas dataset was employed to validate the expression and prognostic significance of FAM83A in women. A small interfering RNA (siRNA) was used for knockdown of the FAM83A gene in HeLa and C33a cells. Next, 5-ethynyl-2&#39;-deoxyuridine (EdU) staining was conducted to determine the effects on the proliferation capabilities of the tumor cells. Wound healing and porous membrane insert assays were performed to evaluate tumor cell migration and invasion abilities.
Western blotting was used to quantify apoptosis-related protein levels. JC-1 staining was employed to evaluate mitochondrial function alterations. Furthermore, cisplatin (diaminedichloroplatinum, DDP) intervention was used to assess the therapeutic potential of the target gene. Flow cytometry and colony formation assays were conducted to further validate the anticancer characteristics of the gene. As a result, FAM83A knockdown was shown to inhibit the proliferation, migration, and invasion of cervical cancer cells and sensitize these cells to cisplatin. These comprehensive methodologies collectively validate FAM83A as a tumor-associated target gene, holding promise as a potential therapeutic target in the prevention and treatment of cervical cancer.

作者聚焦：探索 FAM83A 在宫颈癌中的作用 

Here, we show the procedures for FAM83A knockdown; the assays to detect its effects on proliferation, migration, and invasion of cervical cancer cells; and the sensitization of these cells to cisplatin. This study provides a promising target gene for cervical cancer and a reference for further drug research.

敲低 FAM83A 以验证其在宫颈癌细胞生长和顺铂敏感性中的作用

The exploration of tumor target genes holds paramount importance for the prevention and treatment of cervical cancer. In this study, we outline the steps ...

Through the Looking Glass: Time-lapse Microscopy and Longitudinal Tracking of Single Cells to Study Anti-cancer Therapeutics

The response of single cells to anti-cancer drugs contributes significantly in determining the population response, and therefore is a major contributing factor in the overall outcome. Immunoblotting, flow cytometry and fixed cell experiments are often used to study how cells respond to anti-cancer drugs. These methods are important, but they have several shortcomings. Variability in drug responses between cancer and normal cells, and between cells of different cancer origin, and transient and rare responses are difficult to understand using population averaging assays and without being able to directly track and analyze them longitudinally. The microscope is particularly well suited to image live cells. Advancements in technology enable us to routinely image cells at a resolution that enables not only cell tracking, but also the observation of a variety of cellular responses. We describe an approach in detail that allows for the continuous time-lapse imaging of cells during the drug response for essentially as long as desired, typically up to 96 hr. Using variations of the approach, cells can be monitored for weeks. With the employment of genetically encoded fluorescent biosensors numerous processes, pathways and responses can be followed. We show examples that include tracking and quantification of cell growth and cell cycle progression, chromosome dynamics, DNA damage, and cell death. We also discuss variations of the technique and its flexibility, and highlight some common pitfalls.

Here, we describe a method of long-term time-lapse microscopy to longitudinally track single cells in response to anti-cancer therapeutics.

透过玻璃看:时间推移显微镜和单细胞的纵向跟踪研究抗癌治疗

The response of single cells to anti-cancer drugs contributes significantly in determining the population response, and therefore is a major contributing ...

Using Mouse Mammary Tumor Cells to Teach Core Biology Concepts: A Simple Lab Module

Undergraduate biology students are required to learn, understand and apply a variety of cellular and molecular biology concepts and techniques in preparation for biomedical, graduate and professional programs or careers in science. To address this, a simple laboratory module was devised to teach the concepts of cell division, cellular communication and cancer through the application of animal cell culture techniques. Here the mouse mammary tumor (MMT) cell line is used to model for breast cancer. Students learn to grow and characterize these animal cells in culture and test the effects of traditional and non-traditional chemotherapy agents on cell proliferation. Specifically, students determine the optimal cell concentration for plating and growing cells, learn how to prepare and dilute drug solutions, identify the best dosage and treatment time course of the antiproliferative agents, and ascertain the rate of cell death in response to various treatments. The module employs both a standard cell counting technique using a hemocytometer and a novel cell counting method using microscopy software. The experimental procedure lends to open-ended inquiry as students can modify critical steps of the protocol, including testing homeopathic agents and over-the-counter drugs. In short, this lab module requires students to use the scientific process to apply their knowledge of the cell cycle, cellular signaling pathways, cancer and modes of treatment, all while developing an array of laboratory skills including cell culture and analysis of experimental data not routinely taught in the undergraduate classroom.

A feasible laboratory module for biology undergraduates that explores advanced cellular and molecular concepts using animal cell culture is described. Students grow, characterize and manipulate a breast cancer cell model by exposure to chemotherapy agents. Cell viability is assayed through cell counting using both a standard and novel method.

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

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合成与阿司匹林富马酸酯前药的表征抑制NFκB活动和乳腺癌干细胞

交付使用表皮生长因子在人胰腺癌细胞转染基因受体有针对性的明胶基于工程Nanovectors

细胞凋亡逆转后新形成乳腺癌干细胞样细胞的流式细胞仪检测

B7-H1（PD-L1）免疫组化染色石蜡的嵌入式幻灯片的胰腺癌组织

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

敲低 FAM83A 以验证其在宫颈癌细胞生长和顺铂敏感性中的作用

敲低 FAM83A 以验证其在宫颈癌细胞生长和顺铂敏感性中的作用

敲低 FAM83A 以验证其在宫颈癌细胞生长和顺铂敏感性中的作用

透过玻璃看:时间推移显微镜和单细胞的纵向跟踪研究抗癌治疗

用小鼠乳腺肿瘤细胞教生物学的核心概念:一个简单的实验模块