Name: analysis of combinatorial mirna treatments to regulate cell cycle and angiogenesis
Uploaded: 2019-03-30T14:00:00
Description: Watch this Scientific Journal Video about Analysis of Combinatorial miRNA Treatments to Regulate Cell Cycle and Angiogenesis at JoVE.com

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1. Mir-143 和 Mir-506 转染
注意: 在进行上述实验时, 请使用乳胶手套、防护眼镜和实验室外套。必要时, 请使用带机柜风扇的生物安全柜, 不要堵塞气道或干扰层流。如制造商所述, 始终将保护玻璃窗设置为适当的高度。
<ol><li>种子 NSCLC A549 细胞在 t25 厘米2 flask 96 井板在 dmm/f12k 介质中补充10% 的 fbs 和1% 的青霉素链霉素 (培养培养基) 在组织培养罩中, 在37°c 孵育, 在组织培...

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Mirna 可以作为癌症治疗的靶向疗法, 认识到患病组织与正常组织表达水平的失调。这项研究旨在确定在多个阶段可能阻止细胞周期进展的 Mirna。经证实, Mir-143 和 Mir-506 停止了癌细胞的细胞周期, 并提出了旨在了解这种组合 miRNA 治疗的活性的协议。
所述方法为 Mirna 的功能提供了全面的理解。研究 Mirna 的挑战与它们针对多个基因的能力有关, 因此影响多个途径。所描述的 qPCR 分?...

An erratum was issued for:&#160;<a href="https://www.jove.com/video/59460/analysis-combinatorial-mirna-treatments-to-regulate-cell-cycle">Analysis of Combinatorial miRNA Treatments to Regulate Cell Cycle and Angiogenesis</a>.&#160; An author name was updated.
One of the authors&#39; names was corrected from:
George Matthaiolampakis
to:
George Mattheolabakis

An erratum was issued for: <a href="https://www.jove.com/video/59460/analysis-combinatorial-mirna-treatments-to-regulate-cell-cycle">Analysis of Combinatorial miRNA Treatments to Regulate Cell Cycle and Angiogenesis</a>.&#160; An author name was updated.

Erratum: Analysis of Combinatorial miRNA Treatments to Regulate Cell Cycle and Angiogenesis

细胞周期是多个调控事件的组合, 允许 DNA 复制和细胞通过有丝分裂过程1增殖。环素依赖性激酶 (Cdk) 调节和促进细胞周期 2。其中有丝分裂 CDK (CDK1) 和间期 Cdk (CDK2、CDK4 和 CDK6) 在细胞周期进展3中起着举足轻重的作用。视网膜母细胞瘤蛋白 (Rb) 由 cdkk! cdk6 复合物磷酸化, 以允许细胞周期进展4, CDK1 激活是成功的细胞分裂5的关键。在过去几十年里, 在临床试验中开发并评估了许多 CDK 抑制剂, 这表明在癌症治疗中有可能以 Cdk 为目标。事实上, 最近已经批准了三种 cdk 抑制剂来治疗乳腺癌6、7、8、9、10.因此, Cdk, 特别是 CDK1 和 CDK--在调节癌细胞进展方面非常感兴趣。
Mirna (miRs) 是小型、非编码的 Rna 和转录后基因表达的调节剂, 约占所有人类基因的 30%,11。他们的活动是基于对信使 Rna (Mrna)12的翻译压制或退化。在生物意义的说明中, 已经确定了超过 5, 000个 mirna, 一个单一的 mirna 分子可以调节多个基因11,13。更重要的是, miRNA 的表达与不同的疾病和疾病状态有关, 包括癌症13。事实上, mirna 已被描述为致癌或肿瘤抑制剂, 能够促进或抑制肿瘤的发展和进展14,15。Mirna 在病变组织中的相对表达可以调节疾病的进展;因此, 外源性传递 Mirna 具有治疗潜力。
肺癌是导致癌症相关死亡的主要原因, 超过60% 的肺癌是非小细胞肺癌 16,17 例, 5年生存率不到 20%18.Mirr-143-3p 和 Mir-506-3p 的使用最近被评估为针对肺癌细胞11的细胞周期.mir-143 和 Mir-506 具有与 CDK1 和 cdkkkh/cdk6 互补的序列, 并分析了这两个 miRs 对 A549 细胞的影响。本文对实验细节进行了介绍和讨论。基因表达、细胞周期进展和凋亡的评估使用不同的实验设计和转染后的时间点。我们使用实时定量 PCR (RT-qPCR) 方法和微阵列分析来测量特定的基因表达, 并使用下一代 RNA 测序来确定全球基因失调11。后一种方法以高灵敏度和可重复性识别每个基因转录的相对丰度, 而从单一的实验分析中可以分析数千个基因。此外, 还进行了由于 miRNA 治疗引起的凋亡分析, 并在此进行了描述。生物信息学补充了路径分析。这里提出的协议用于分析组合 Mir-143 和 Mir-506 的治疗潜力。
该协议的主要目的是确定 Mirna 在细胞中的影响, 重点是细胞周期。这里介绍的各种技术从基因表达分析翻译前 (使用 qPCR) 到在蛋白质水平上详细阐述和新的基因分析技术, 如微阵列分析。希望这份报告对有兴趣与 Mirna 合作的研究人员有所帮助。此外, 还提出了细胞周期和细胞凋亡的流式细胞仪分析方法。

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		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:519px;">Eppendorf Pipette Pick-A-Pack Sets</td>
			<td style="width:210px;">Eppendrof</td>
			<td style="width:1167px;">05-403-152</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Ethanol, Absolute (200 Proof), Molecular Biology Grade,&#160;</td>
			<td style="width:210px;">Fisher BioReagents</td>
			<td>BP2818500</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:519px;">Ethidium bromide</td>
			<td style="width:210px;">Alfa acar</td>
			<td style="width:1167px;">L07462</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">F-12K Nutrient Mixture (Kaighn&#39;s Mod.) with L-glutamine, Corning</td>
			<td style="width:210px;">Corning</td>
			<td>45000-354</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:519px;">FACS Calibur Flowcytometer</td>
			<td style="width:210px;">Becton Dickinson</td>
			<td style="width:1167px;"></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Fetal Bovine Serum - Premium</td>
			<td>Antlanta Biologicals</td>
			<td>S11150</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:519px;">Fetal Bovine Serum (FBS)</td>
			<td style="width:210px;">Fisher Scientific</td>
			<td style="width:1167px;">10438026</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Fisherbrand Basix Microcentrifuge Tubes with Standard Snap Caps</td>
			<td style="width:210px;">Fisherbrand Basix</td>
			<td style="width:1167px;">02-682-002</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:519px;">Forma Series II water Jacket CO2 incubator</td>
			<td style="width:210px;">Thermo Scientific</td>
			<td style="width:1167px;"></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:519px;">Heparin Solution (5000 U/mL)</td>
			<td style="width:210px;">Hospira</td>
			<td style="width:1167px;">NDC#63739-920-11</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:519px;">Horixontal Electrophoresis system</td>
			<td style="width:210px;">Benchtop lab system</td>
			<td style="width:1167px;">BT102</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">hsa-miR-143-3p miRNA Mimic</td>
			<td style="width:210px;">ABM</td>
			<td>MCH01315</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">hsa-miR-506-3p miRNA Mimic</td>
			<td style="width:210px;">ABM</td>
			<td>MCH02824</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:519px;">Human Recombinant Vascular Endothelial Growth Factor (VEGF)</td>
			<td style="width:210px;">Thermo Scientific</td>
			<td style="width:1167px;">PHC9394&#160;&#160;</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:519px;">Human Umbilical Vein Endothelial Cells (HUVEC)</td>
			<td style="width:210px;">Individual donors</td>
			<td style="width:1167px;">IRB# A15-3891</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:519px;">HyClone Phosphate Buffered Saline (PBS)</td>
			<td style="width:210px;">Fisher Scientific</td>
			<td style="width:1167px;">SH30256FS</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:519px;">Ingenuity Pathway Analysis</td>
			<td style="width:210px;">Qiagen</td>
			<td style="width:1167px;">Results: Used for bioinformatics pathway analysis</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Invitrogen UltraPure DNase/RNase-Free Distilled Water</td>
			<td style="width:210px;">Invitrogen</td>
			<td>10-977-015</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:519px;">Lipofectamine 2000&#160;</td>
			<td style="width:210px;">Invitrogen</td>
			<td style="width:1167px;">11-668-027</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:519px;">Loading dye 10X</td>
			<td style="width:210px;">ward&#39;s science+</td>
			<td style="width:1167px;">470024-814</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:519px;">Medium M199 (with Earle&#8242;s salts, L-glutamine and sodium bicarbonate)</td>
			<td style="width:210px;">Sigma Aldrich</td>
			<td style="width:1167px;">M4530</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:519px;">Microscope Digital Camera</td>
			<td style="width:210px;">AmScope&#160;</td>
			<td style="width:1167px;">MU130</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:519px;">Modfit LT</td>
			<td style="width:210px;">Verity Software</td>
			<td style="width:1167px;">Step 5.15: Alternative software for analysis of cell cycle population distributions</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:519px;">Nanodrop</td>
			<td style="width:210px;">Thermo Scientific</td>
			<td style="width:1167px;">NanoDrop one C</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Opti-MEM</td>
			<td style="width:210px;">Gibco by life technologies</td>
			<td>31985-070</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Penicillin-streptomycin 10/10</td>
			<td>Antlanta Biologicals</td>
			<td>B21210</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Power UP sybr green master mix</td>
			<td style="width:210px;">Applied Biosystems</td>
			<td>A25780</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Propidium Iodide</td>
			<td style="width:210px;">MP Biochemicals LLC</td>
			<td>IC19545825</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:519px;">Proscanarray HT Microarray scanner</td>
			<td style="width:210px;">Perkin elmer</td>
			<td style="width:1167px;">ASCNPHRG. We used excitation laser wavelength at 543 nm.</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">q PCR optical adhesive cover</td>
			<td style="width:210px;">Applied Biosystems</td>
			<td style="width:1167px;">4360954</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:519px;">Quick-RNA Kits</td>
			<td>Zymo Research</td>
			<td>R1055</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Ribonuclease A from Bovine pancreas</td>
			<td style="width:210px;">Sigma</td>
			<td>R6513-50MG</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:519px;">ScanArray Express</td>
			<td>PerkinElmer</td>
			<td style="width:1167px;">Step 7.33: Microarray analysis software</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:519px;">Shaker</td>
			<td style="width:210px;">Thermo Scientific</td>
			<td style="width:1167px;">2314</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:519px;">SimpliAmp Thermal Cycler</td>
			<td style="width:210px;">Applied Biosystems</td>
			<td style="width:1167px;"></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:519px;">SpectraTube Centrifuge Tubes 15ml</td>
			<td style="width:210px;">VWR</td>
			<td style="width:1167px;">470224-998</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:519px;">SpectraTube Centrifuge Tubes 50ml</td>
			<td style="width:210px;">VWR</td>
			<td style="width:1167px;">470225-004</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">TBS Buffer, 20x liquid</td>
			<td style="width:210px;">VWR</td>
			<td>10791-796</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:519px;">Temperature controlled&#160; centrifuge matchine</td>
			<td style="width:210px;">Thermo Scientific</td>
			<td style="width:1167px;">ST16R</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:519px;">Temperature controlled micro centrifuge matchine</td>
			<td style="width:210px;">Eppendrof</td>
			<td style="width:1167px;">5415R</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Thermo Scientific BioLite Cell Culture Treated Flasks</td>
			<td>Thermo Scientific</td>
			<td>12-556-009</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:519px;">Thermo Scientific Pierce BCA Protein Assay</td>
			<td style="width:210px;">Thermo Scientific</td>
			<td style="width:1167px;">PI23225</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Thermo Scientific Pierce RIPA Buffer</td>
			<td style="width:210px;">Thermo Scientific</td>
			<td>PI89900</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Thermo Scientific Thermo-Fast 96-Well Full-Skirted Plates</td>
			<td style="width:210px;">Thermo Scientific</td>
			<td style="width:1167px;">AB0800WL</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Thermo Scientific Verso cDNA synthesis Kit (100 runs)</td>
			<td style="width:210px;">Thermo Scientific</td>
			<td>AB1453B</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:519px;">Ultra Low Range DNA Ladder</td>
			<td style="width:210px;">Invitrogen</td>
			<td style="width:1167px;">10597012</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:519px;">VWR standard solid door laboratory refrigerator</td>
			<td style="width:210px;">VWR</td>
			<td style="width:1167px;"></td>
			<td></td>
		</tr>
	</tbody>
</table>

analysis of combinatorial mirna treatments to regulate cell cycle and angiogenesis

肺癌 (LC) 是全球癌症相关死亡的主要原因。与其他癌细胞类似, LC 细胞的一个基本特征是不受管制的增殖和细胞分裂。通过阻止细胞周期进展抑制增殖已被证明是一种有前途的癌症治疗方法, 包括 LC。
miRNA 疗法已成为重要的转录后基因调节剂, 并越来越多地被研究用于癌症治疗。在最近的工作中, 我们使用了两个 Mirna, Mir-143 和 Mir-506, 以调节细胞周期的进展。对 A549 非小细胞肺癌 (NSCLC) 细胞进行转染, 分析基因表达改变, 最后分析治疗后的凋亡活性。检测到环素依赖性激酶 (cdk) 的下行 (即 CDK1、CDK4 和 CDK6) 的下行调节, 并在 g试管和 g2m 相变时停止细胞周期。病理分析显示治疗具有潜在的抗血管生成活性, 这使得该方法具有多方面的活性。这里, 描述了用于识别 miRNA 活性有关细胞周期抑制, 诱导细胞凋亡, 以及通过抑制血管生成治疗对内皮细胞的影响的方法。希望这里介绍的方法将支持未来对 miRNA 疗法和相应活性的研究, 并希望有代表性的数据将指导其他研究人员进行实验分析。

RT-qPCR 和凝胶电泳基因表达分析
利用 RT-qPCR 进行的差异基因表达分析显示, 靶向基因 CDK1、CDK4 和 CDK6 有显著下调。CDK1 和 cdkk 6 分别有助于 g2m 和 GN.S 的过渡。所进行的分析可以直接比较单个 miRs 和组合 MiRs 活动。利用对检测到的基因下调过程中的任何干扰进行检测, 使其与转染剂的制备, 是最小的。使用双尾学生的...

Watch this Scientific Journal Video about Analysis of Combinatorial miRNA Treatments to Regulate Cell Cycle and Angiogenesis at JoVE.com

Analysis of Combinatorial miRNA Treatments to Regulate Cell Cycle and Angiogenesis

miRNA 疗法在调节癌症进展方面具有巨大的潜力。这里演示了用于鉴定组合 miRNA 治疗在停止细胞周期和血管生成方面的活性的分析方法。

组合 miRNA 治疗调节细胞周期和血管生成的分析

Lung cancer (LC) is the leading cause of cancer-related deaths worldwide. Similar to other cancer cells, a fundamental characteristic of LC cells is ...

此方法将有助于理解和执行必要的实验室技术，以确定两个微RNA在体外肺癌细胞中的活性。肺癌是全世界癌症相关死亡的主要原因。在这里，我们介绍从基本实验室方法到更复杂的协议的技术，如基因表达分析和抗体蛋白微RNA，专门用于识别微RNA的作用。以下方法将帮助我们回答有关两个微RNA，微RNA143和微RNA506对调节肺癌细胞周期的影响的关键问题。我们提出了不同的实验室技术，如细胞转染、RNA提取、定量实时PCR和微RNA分析。我们希望我们的介绍将有助于成功完成这些分析。首先，将培养的细胞安占一个足以提取RNA和蛋白质的适当烧瓶中，使用qPCR或微阵列分析进行基因表达。然后用10%的FBS和1%的青霉素链霉素补充一夜的孵化。第二天，在介质中稀释微RNA 143和微RNA 506，制备微RNA样本，每个浓度为100纳米摩尔。从培养箱中取出烧瓶并取出介质。用 1x PBS 清洗。所有未经处理的对控将只包含与细胞的不完整介质。用转染培养库孵化细胞6小时。6 小时后，取出介质，更换四毫升新鲜、完整的介质。24小时48小时后通过三辛化收获细胞。接下来，在每个管中用1x PBS清洗两次，然后取出上一根。为了在以后进行RNA提取，在零下80摄氏度时冻结细胞。RNA提取是按照制造商的指示使用RNA试剂盒进行的。首先，从零下80摄氏度的温度下取出管子，并允许解冻。加入300微升的解液缓冲液和移液器上下，以分解细胞膜。加入100%超纯乙醇的同等体积。然后混合好，放在分开的列中。离心并拆经。添加 400 微升的洗涤和缓冲液以及离心机以去除缓冲液。在每个样品中加入5微升DNAse 1，加入75微升VNA消化缓冲液，孵育15分钟。然后用400微升RNA制备缓冲液清洗样品。接下来，用RNA洗涤缓冲液洗两次。现在，将无核酸酶水加入柱，离心机，然后收集RNA。测量RNA浓度。在这个阶段，可以发送两微克RNA样本用于RNA测序。使用热循环器，根据文本部分中描述的协议准备 cDNA。使用 DNAse RNAse 无水准备前向和反向底转解决方案，浓度为 10 微摩尔。准备根据样本数检测的每个基因的主混合。对于每个 cDNA 样品，反应量根据文本中描述的表。将每个样品放在各自的井中。对于 qPCR，准备具有 96 井板布局的 Excel 文档总是好的，以便跟踪样本。对于每个样本和分析的基因，执行其反应的三元，或至少重复。使用透明、光学透明的塑料密封器密封塑料板。避免用手指触摸薄塑料层，因为这会产生错误信号。快速旋转板，将所有试剂混合到井底。接下来，使用定量的实时 PCR 机运行样品板。获取 qPCR 机生成的 CT 值，并分析相关基因表达。如本视频前面部分所述，对细胞进行转染。从每个样品中收获细胞，在单独的15毫升无菌管中进行三辛化。接下来，用1x PBS清洗细胞两次，在751G下离心5分钟，然后取出上经。通过移液器添加 200 微升冰冷 1x PBS，重新挂起并打破调色板。加入两毫升70%的冰冷乙醇，滴到管子上，同时轻轻旋转管子来修复细胞。在室温下孵育管30分钟，并在4摄氏度内放置1小时。从四摄氏度和离心机取出管子。加入2ml冰冷的PBS和涡流，然后通过离心去除上清液。在每个样品中加入500微升含有碘化二和核糖核酸A的1x PBS，浓度为每毫升50微克和200微克，并在室温下孵育30分钟。将样品转移到适当的管中，防止光线，并在流动环流仪上运行。本实验是识别所有细胞周期通路蛋白的表达，通过微RNA治疗改变。根据转染部分所述程序收集蛋白质样本，使用 BCA 测定进行量化。在实验前一小时从零下四摄氏度的温度下取下滑梯，使室温升高并去除水分。采集 70 微克蛋白质样本，根据制造商的说明准备样品幻灯片，在文本部分也分步描述。请注意，用超纯脱水正确清洗幻灯片是另一个重要步骤，因为这将有助于减少幻灯片的背景。此外，在此实验中，直到最后一步才将示例幻灯片干涸，这一点非常关键。最后，使用微阵列机扫描幻灯片并分析数据。对微RNA处理样品进行qPCR分析，以确定CDK1、CDK4和CDK6的基因表达。这些基因调节正常细胞和癌细胞的细胞周期，并已被作为抑制肿瘤进展的手段。数据表明，由于对微RNA143和506的治疗，这些基因的调节作用降低。来自流细胞学的细胞周期分布数据表明，由于微RNA143和506组合处理的影响，G0和G1阶段的种群数量增加，S相的种群减少。细胞周期的特定微阵列分析检测出大约60个基因的蛋白质表达变化。表示的热图显示细胞周期进展和增殖所需的多个基因的蛋白质水平的降低调节，如CDK2、海伦1和3、RE2F1。相比之下，由其生长的抑制作用所识别的蛋白质被调高。虽然结果为半定量，需要进一步分析，但微阵列分析提供了路径活性的快速概述。我们相信，看完这段视频后，您将能够进行基本的实验室实验，帮助您识别细胞中的微RNA活性。

Research

JoVE Journal

Cancer Research

MicroRNA Based Liquid Biopsy: The Experience of the Plasma miRNA Signature Classifier (MSC) for Lung Cancer Screening

MicroRNA Based Liquid Biopsy: The Experience of the Plasma miRNA Signature Classifier MSC for Lung Cancer Screening

基于 rna 的液体活检: 血浆 miRNA 特征分类器 (MSC) 在肺癌筛查中的经验

The development of a minimally invasive test, such as liquid biopsy, for early lung cancer detection in its preclinical phase is crucial to improve the ...

科研

癌症研究

Preparation of Primary Acute Lymphoblastic Leukemia Cells in Different Cell Cycle Phases by Centrifugal Elutriation

离心淘洗制备不同细胞周期的原发性急性淋巴细胞白血病细胞

The ability to synchronize cells has been central to advancing our understanding of cell cycle regulation. Common techniques employed include serum ...

Simple and Rapid Method to Obtain High-quality Tumor DNA from Clinical-pathological Specimens Using Touch Imprint Cytology

用触印细胞学方法快速获得临床病理标本中高质量肿瘤 DNA

It is critical to determine the mutational status in cancer before administration and treatment of specific molecular targeted drugs for cancer patients. ...

Generation and Isolation of Cell Cycle-arrested Cells with Complex Karyotypes

复杂核型细胞周期捕获细胞的生成与分离

Chromosome mis-segregation leads to aneuploidy, a condition in which cells harbor an imbalanced chromosome number. Several lines of evidence strongly ...

Phospho Flow Cytometry with Fluorescent Cell Barcoding for Single Cell Signaling Analysis and Biomarker Discovery

磷流式细胞仪与荧光细胞条形码用于单细胞信号分析和生物标志物发现

Aberrant cell signaling plays a central role in cancer development and progression. Most novel targeted therapies are indeed directed at proteins and ...

Flow Cytometric Analysis of Mitochondrial Reactive Oxygen Species in Murine Hematopoietic Stem and Progenitor Cells and MLL-AF9 Driven Leukemia

木尿造血干细胞和祖细胞和MLL-AF9驱动白血病线粒体反应氧物种的流细胞学分析

We present a flow cytometric approach for analyzing mitochondrial ROS in various live bone marrow (BM)-derived stem and progenitor cell populations from ...

Optimization, Design and Avoiding Pitfalls in Manual Multiplex Fluorescent Immunohistochemistry

手动复用荧光免疫学的优化、设计和避免陷阱

Microenvironment evaluation of intact tissue for analysis of cell infiltration and spatial organization are essential in understanding the complexity of ...

Cell Cycle-specific Measurement of &#947;H2AX and Apoptosis After Genotoxic Stress by Flow Cytometry

细胞周期特异性测量βH2AX和细胞凋亡后热源毒性应力通过流细胞测定

The presented method or slightly modified versions have been devised to study specific treatment responses and side effects of various anti-cancer ...

Analysis of Side Population in Solid Tumor Cell Lines

固体肿瘤细胞系侧面人口分析

Cancer stem cells (CSCs) are an important cause of tumor growth, metastasis, and recurrence. Isolation and identification of CSCs are of great ...

Evaluating Cell Death Using Cell-Free Supernatant of Probiotics in Three-Dimensional Spheroid Cultures of Colorectal Cancer Cells

在结肠直肠癌细胞的三维球体培养物中使用无细胞益生菌超常剂评估细胞死亡

This manuscript describes a protocol to evaluate cancer cell deaths in three dimensional (3D) spheroids of multicellular types of cancer cells using ...