Name: establishing dual resistance to egfr-tki and met-tki in lung adenocarcinoma cells in vitro with a 2-step dose-escalation procedure
Uploaded: 2017-08-11T14:00:00
Description: Watch this Scientific Journal Video about Establishing Dual Resistance to EGFR-TKI and MET-TKI in Lung Adenocarcinoma Cells In Vitro with a 2-step Dose-escalation Procedure at JoVE.com

我们感谢他们提出深思熟虑的意见和有益的建议，分子肿瘤研究所和 Editage 提供的援助与英语语言编辑成员。这项工作是为私立大学 （2012年-2016 年） 从教育部、 文化、 体育、 科学和技术的日本战略研究基金会支持由文部科学省支持程序。钱 Ohmori 收到勃林格殷格翰 （殷格翰，德国） 资金 （2015年-2016 年） 的研究。

1.建立吉非替尼耐药细胞
<ol>
	<li>3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium 溴 (MTT) 法测定确定初始吉非替尼治疗浓度。
	<ol>
		<li>细胞生长介质中的 PC-9 (rpmi-1640 培养基用 10%胎牛血清、 1%抗生素 (青霉素： 100ml U 和链霉素： 100 微克/毫升))，在一个 5%CO2的孵化器在 37 ° c。</li>
		<li>调整单元格到 1.0 × 105细胞/毫升使用自动化的单元反击 （见材料表） 和种子 5...

<ol>
	<li>Lynch, T. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Activating+mutations+in+the+epidermal+growth+factor+receptor+underlying+responsiveness+of+non-small-cell+lung+cancer+to+gefitinib.">Activating mutations in the epidermal growth factor receptor underlying responsiveness of non-small-cell lung cancer to gefitinib.</a> N Engl J Med. 350 (21), 2129-2139 (2004).</li><li>Paez, J. G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=EGFR+mutations+in+lung+cancer:+correlation+with+clinical+response+to+gefitinib+therapy.">EGFR mutations in lung cancer: correlation with clinical response to gefitinib therapy.</a> Science. 304 (5676), 1497-1500 (2004).</li><li>Pao, W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=EGF+receptor+gene+mutations+are+common+in+lung+cancers+from+"never+smokers"+and+are+associated+with+sensitivity+of+tumors+to+gefitinib+and+erlotinib.">EGF receptor gene mutations are common in lung cancers from "never smokers" and are associated with sensitivity of tumors to gefitinib and erlotinib.</a> Proc Natl Acad Sci U S A. 101 (36), 13306-13311 (2004).</li><li>Mitsudomi, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mutations+of+the+epidermal+growth+factor+receptor+gene+predict+prolonged+survival+after+gefitinib+treatment+in+patients+with+non-small-cell+lung+cancer+with+postoperative+recurrence.">Mutations of the epidermal growth factor receptor gene predict prolonged survival after gefitinib treatment in patients with non-small-cell lung cancer with postoperative recurrence.</a> J Clin Oncol. 23 (11), 2513-2520 (2005).</li><li>Sequist, L. V., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genotypic+and+histological+evolution+of+lung+cancers+acquiring+resistance+to+EGFR+inhibitors.">Genotypic and histological evolution of lung cancers acquiring resistance to EGFR inhibitors.</a> Sci Transl Med. 3 (75), 75ra26 (2011).</li><li>Engelman, J. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=MET+amplification+leads+to+gefitinib+resistance+in+lung+cancer+by+activating+ERBB3+signaling.">MET amplification leads to gefitinib resistance in lung cancer by activating ERBB3 signaling.</a> Science. 316 (5827), 1039-1043 (2007).</li><li>Schmidt, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Novel+mutations+of+the+MET+proto-oncogene+in+papillary+renal+carcinomas.">Novel mutations of the MET proto-oncogene in papillary renal carcinomas.</a> Oncogene. 18 (14), 2343-2350 (1999).</li><li>Olivero, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Novel+mutation+in+the+ATP-binding+site+of+the+MET+oncogene+tyrosine+kinase+in+a+HPRCC+family.">Novel mutation in the ATP-binding site of the MET oncogene tyrosine kinase in a HPRCC family.</a> Int J Cancer. 82 (5), 640-643 (1999).</li><li>Janne, P. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=AZD9291+in+EGFR+inhibitor-resistant+non-small-cell+lung+cancer.">AZD9291 in EGFR inhibitor-resistant non-small-cell lung cancer.</a> N Engl J Med. 372 (18), 1689-1699 (2015).</li><li>Bean, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=MET+amplification+occurs+with+or+without+T790M+mutations+in+EGFR+mutant+lung+tumors+with+acquired+resistance+to+gefitinib+or+erlotinib.">MET amplification occurs with or without T790M mutations in EGFR mutant lung tumors with acquired resistance to gefitinib or erlotinib.</a> Proc Natl Acad Sci U S A. 104 (52), 20932-20937 (2007).</li><li>. NCT02468661 - ClinicalTrials.gov Available from: <a target="_blank" href="https://clinicaltrials.gov/ct2/show/NCT02468661">https://clinicaltrials.gov/ct2/show/NCT02468661</a> (2017)</li><li>Yamaoka, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acquired+Resistance+Mechanisms+to+Combination+Met-TKI/EGFR-TKI+Exposure+in+Met-Amplified+EGFR-TKI-Resistant+Lung+Adenocarcinoma+Harboring+an+Activating+EGFR+Mutation.">Acquired Resistance Mechanisms to Combination Met-TKI/EGFR-TKI Exposure in Met-Amplified EGFR-TKI-Resistant Lung Adenocarcinoma Harboring an Activating EGFR Mutation.</a> Mol Cancer Ther. 15 (12), 3040-3054 (2016).</li><li>Mosmann, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rapid+colorimetric+assay+for+cellular+growth+and+survival:+application+to+proliferation+and+cytotoxicity+assays.">Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays.</a> J Immunol Methods. 65 (1-2), 55-63 (1983).</li><li>Cifone, M. A., Fidler, I. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Correlation+of+patterns+of+anchorage-independent+growth+with+in+vivo+behavior+of+cells+from+a+murine+fibrosarcoma.">Correlation of patterns of anchorage-independent growth with in vivo behavior of cells from a murine fibrosarcoma.</a> Proc Natl Acad Sci U S A. 77 (2), 1039-1043 (1980).</li><li>Ercan, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Amplification+of+EGFR+T790M+causes+resistance+to+an+irreversible+EGFR+inhibitor.">Amplification of EGFR T790M causes resistance to an irreversible EGFR inhibitor.</a> Oncogene. 29 (16), 2346-2356 (2010).</li><li>Ogino, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Emergence+of+epidermal+growth+factor+receptor+T790M+mutation+during+chronic+exposure+to+gefitinib+in+a+non+small+cell+lung+cancer+cell+line.">Emergence of epidermal growth factor receptor T790M mutation during chronic exposure to gefitinib in a non small cell lung cancer cell line.</a> Cancer Res. 67 (16), 7807-7814 (2007).</li><li>Ando, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Enhancement+of+sensitivity+to+tumor+necrosis+factor+alpha+in+non-small+cell+lung+cancer+cells+with+acquired+resistance+to+gefitinib.">Enhancement of sensitivity to tumor necrosis factor alpha in non-small cell lung cancer cells with acquired resistance to gefitinib.</a> Clin Cancer Res. 11 (24 Pt 1), 8872-8879 (2005).</li><li>Suda, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reciprocal+and+complementary+role+of+MET+amplification+and+EGFR+T790M+mutation+in+acquired+resistance+to+kinase+inhibitors+in+lung+cancer.">Reciprocal and complementary role of MET amplification and EGFR T790M mutation in acquired resistance to kinase inhibitors in lung cancer.</a> Clin Cancer Res. 16 (22), 5489-5498 (2010).</li><li>Shien, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acquired+resistance+to+EGFR+inhibitors+is+associated+with+a+manifestation+of+stem+cell-like+properties+in+cancer+cells.">Acquired resistance to EGFR inhibitors is associated with a manifestation of stem cell-like properties in cancer cells.</a> Cancer Res. 73 (10), 3051-3061 (2013).</li><li>Sagawa, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Establishment+of+three+cisplatin-resistant+endometrial+cancer+cell+lines+using+two+methods+of+cisplatin+exposure.">Establishment of three cisplatin-resistant endometrial cancer cell lines using two methods of cisplatin exposure.</a> Tumour Biol. 32 (2), 399-408 (2011).</li><li>Ritter, C. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Human+breast+cancer+cells+selected+for+resistance+to+trastuzumab+in+vivo+overexpress+epidermal+growth+factor+receptor+and+ErbB+ligands+and+remain+dependent+on+the+ErbB+receptor+network.">Human breast cancer cells selected for resistance to trastuzumab in vivo overexpress epidermal growth factor receptor and ErbB ligands and remain dependent on the ErbB receptor network.</a> Clin Cancer Res. 13 (16), 4909-4919 (2007).</li></ol>

这里描述的协议可以用于获得双抗性使用 2 步剂量递增法体外癌细胞中建立。许多研究论文报道，PC-9 细胞抗药性 EGFR-使用 tki 药物治疗 （吉非替尼或厄洛替尼） 通过 T790M EGFR 突变15，16。然而，我们没有检测到 T790M 突变基因外显子 20 的我们的 PC 9MET1000 细胞通过直接测序法 (图 4)。此外，大都会放大观察作为信号转导通路?...

致癌性突变中表皮生长因子受体 (EGFR) 发现在非小细胞肺癌 (NSCLC) 患者的一个子集，重要预测生物标记物此疾病1，2。这些最常用激活 EGFR 突变发生作为帧中删除在 19 号外显子 (delE746-A750) 或替换亮氨酸与精氨酸的外显子 (存在 L858R) 21 号 858 密码在 EGFR 酪氨酸激酶域3，4点突变。窝藏 EGFR 突变的非小细胞肺癌患者的临床有效治疗是治疗 EGFR 酪氨酸激酶抑制剂 （测出）。然而，这种疗法被受到限制抗 EGFR 测出吉非替尼和埃罗替尼等必然采集。最常见的获得性耐药发生，通过二次 T790M 突变基因外显子 20 的 EGFR，大约 60%的患者获得 EGFR-使用 tki 药物治疗 （吉非替尼或厄洛替尼） 电阻检测到。其他伴 EGFR 测出获得性耐药的分子机制并激活的旁路信号引起的大都会放大，转型小细胞肺癌，与上皮-间充质转变5，6。受体肝细胞生长因子，由 MET 基因，编码是 dysregulatedin 许多肿瘤，据报是重要的一个候选靶向疗法7，8。
战略，以克服 EGFR 测出获得性的耐药正进行临床评价。临床前和临床研究表明，第三代 EGFR 抑制剂如 osimertinib 是 T790M 突变9患者有效。此外，可通过联合治疗 EGFR 和大都会测出6，10抑制 EGFR 突变癌症与大都会放大的增长。评估患者的大都会和 EGFR 抑制剂组合与获得性耐药吉非替尼和埃罗替尼的临床试验正在进行现在是11。
研究对化疗药物产生获得性耐药的分子机制，最初响应抑制剂的细胞系是与这些抑制剂继续治疗。两种方法可用来建立获得性的耐药细胞系。一是逐步的剂量递增，和另一种是接触浓度高。逐步升级方法已被更常用，因为它具有很高的成功率。单元格第一次接触低浓度的抑制剂，近十分之一的半-极大抑制浓度 (IC50) 值，和浓度然后逐渐增加，由 10-30%直到达到靶浓度。高质量浓度暴露法涉及细胞暴露于最后一剂到培养基中，是超过 IC50值，所以几乎彻底杀死亲本细胞抑制剂。此高质量浓度暴露方法具有更低成功率比逐步升级方法。
在以往的研究中，EGFR-使用 tki 药物治疗与 MET-使用 tki 药物治疗联合的治疗被证明是有效的体外系统。获得性的耐药 EFGR-使用 tki 药物治疗，吉非替尼通过逐步升级为 12 个月，MET-放大，成立，因此，吉非替尼耐药细胞线称为 PC 9MET1000 生成的12。
该协议是两步剂量升级程序获得双抗 EGFR-使用 tki 药物治疗，吉非替尼和大都会-使用 tki 药物治疗，PHA665752 的 EGFR 突变的非小细胞肺癌 PC 9 细胞 (图 1)。这种方法建立获得性耐药细胞系中是相对较容易执行，成功率高。描述的协议可以修改为其他分子靶向药物的细胞毒性药物以及抑制剂的应用。

<table width="970">
	<tbody>
		<tr height="21">
			<td height="21" style="width:365px;height:21px;">RPMI-1640</td>
			<td style="width:177px;">Wako</td>
			<td style="width:119px;">189-02025</td>
			<td style="width:309px;">with L-Glutamine and Phenol Red</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">CellTiter 96</td>
			<td style="width:177px;">Promega</td>
			<td style="width:119px;">G4100</td>
			<td>Non-Radioactive Cell Proliferation Assay</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:365px;height:21px;">FBS</td>
			<td style="width:177px;">gibco</td>
			<td style="width:119px;">26140-079</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="width:365px;height:21px;">Pen Strep</td>
			<td style="width:177px;">gibco</td>
			<td style="width:119px;">15140-163</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="width:365px;height:21px;">gefitinib</td>
			<td style="width:177px;">Selleck</td>
			<td style="width:119px;">S1025</td>
			<td>stocked 10 mM/EMSO at -20&#186;C</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:365px;height:21px;">PHA665752</td>
			<td style="width:177px;">Selleck</td>
			<td style="width:119px;">S1070</td>
			<td>stocked 10 mM/EMSO at -20&#186;C</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:365px;height:21px;">96-well plate</td>
			<td style="width:177px;">IWAKI</td>
			<td style="width:119px;">860-096</td>
			<td>flat bottom with lid, low evaporation type</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:365px;height:21px;">TC10 Automated Cell Counter</td>
			<td style="width:177px;">BIO RAD</td>
			<td>#1450010</td>
			<td></td>
		</tr>
		<tr height="26">
			<td height="26" style="width:365px;height:26px;">CellTiter 96 Non-Radioactive Cell Proliferation Assay</td>
			<td style="width:177px;">Promega</td>
			<td style="width:119px;">G4100</td>
			<td style="width:309px;">Dye Solution and Solubilization/Stop Solution</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Powerscan HT microplate reader</td>
			<td>BioTek</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="26">
			<td height="26" style="width:365px;height:26px;">GraphPad Prism v.5.0 software</td>
			<td style="width:177px;">GraphPad Inc.</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="26">
			<td height="26" style="width:365px;height:26px;">PC-9</td>
			<td style="width:177px;">Riken BioResource Center</td>
			<td style="width:119px;">RCB4455</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="width:365px;height:21px;">P100 dish</td>
			<td style="width:177px;">FALCON</td>
			<td align="right" style="width:119px;">353003</td>
			<td></td>
		</tr>
	</tbody>
</table>

establishing dual resistance to egfr-tki and met-tki in lung adenocarcinoma cells in vitro with a 2-step dose-escalation procedure

耐药是在癌症治疗中的一项重大挑战。耐药株在体外生成是发现新的机制，以克服这一挑战的必要条件。在这里，一种建立双电阻对表皮生长因子受体 (EGFR) 的 2 步剂量升级方法-酪氨酸激酶抑制剂 （使用 tki 药物治疗）、 吉非替尼和大都会-使用 tki 药物治疗，PHA665752，描述。这种方法基于简单逐步剂量递增抑制剂诱导的获得性抗性细胞系。产生耐药株的备用方法涉及细胞暴露于高浓度抑制剂中的一步。逐步的剂量递增方法具有更高的成功诱导可能性获得性抗性比这种方法。激活 EGFR 突变是响应与 EGFR-使用 tki 药物治疗，治疗的生物标志物应用的一线治疗非小细胞肺癌 (NSCLC)，那怀有这些突变。然而，尽管作出有效反应的报告，EGFR-使用 tki 药物治疗使用是有限因为肿瘤不可避免地获得耐药性。EGFR-使用 tki 药物治疗抵抗背后的主要机制包括看门人网站、 在 EGFR 外显子 20 T790M 和大都会旁路信号二次突变。因此，为这一问题的潜在解决方案将结合 EGFR-使用 tki 药物治疗和大都会-使用 tki 药物治疗。这个综合的治疗已被证明是有效的体外研究模型。获得性吉非替尼耐通过 MET 放大而成立的逐步剂量递增的吉非替尼治疗 12 个月，命名为 PC 9MET1000 细胞株在先前的研究中生成。进一步探讨获得性的 MET-使用 tki 药物治疗和 EGFR-使用 tki 药物治疗的机制抵抗，大都会-使用 tki 药物治疗，PHA665752，管理，给这些细胞逐步剂量递增吉非替尼在为 12 个月。本议定书也成功地适用于组合疗法建立获得性的耐药其他抑制剂分子的数目。

这手稿 》 中提出了议定书 》 概述如图 1，包括对吉非替尼和 PHA665752 的抗性双感应 PC 9 单元格中通过 2 步剂量升级过程所示。图 2显示了细胞增殖的家长 PC 9 随着浓度的增加吉非替尼的跌幅。这表明，PC-9 细胞变得敏感对吉非替尼剂量升级过程完成之后。图 3显示 PC 9MET1000 细胞表现出电阻对吉非替尼，但?...

Watch this Scientific Journal Video about Establishing Dual Resistance to EGFR-TKI and MET-TKI in Lung Adenocarcinoma Cells In Vitro with a 2-step Dose-escalation Procedure at JoVE.com

Establishing Dual Resistance to EGFR-TKI and MET-TKI in Lung Adenocarcinoma Cells In Vitro with a 2-step Dose-escalation Procedure

在体外建立双抗 EGFR-使用 tki 药物治疗和癌症细胞中大都会使用 tki 药物治疗方法。此方法是用于开发陈列尽管大都会放大 EGFR-使用 tki 药物治疗治疗疾病进展的 EGFR 突变，患者的治疗方法。它也可以修改为针对其他分子的抑制剂。

建立双抗 EGFR-使用 tki 药物治疗和满足使用 tki 药物治疗在肺腺癌细胞体外与 2 步剂量升级程序

Drug resistance is a major challenge in cancer therapy. The generation of resistant sublines in vitro is necessary for discovering novel mechanisms to ...

Research

JoVE Journal

Cancer Research

Establishing Dual Resistance to EGFR-TKI and MET-TKI in Lung Adenocarcinoma Cells In Vitro with a 2-step Dose-escalation Procedure

Long-term High-Resolution Intravital Microscopy in the Lung with a Vacuum Stabilized Imaging Window

长期高分辨率活体显微镜在肺与真空稳定成像窗口

Metastasis to secondary sites such as the lung, liver and bone is a traumatic event with a mortality rate of approximately 90% 1. Of these sites, the lung ...

科研

癌症研究

Using RNA-sequencing to Detect Novel Splice Variants Related to Drug Resistance in In Vitro Cancer Models

Using RNA-sequencing to Detect Novel Splice Variants Related to Drug Resistance in In Vitro Cancer Models

利用RNA测序来检测新剪接变异体在相关耐药<em&gt;体外</em&gt;癌症模型

Drug resistance remains a major problem in the treatment of cancer for both hematological malignancies and solid tumors. Intrinsic or acquired resistance ...

Moving Upwards: A Simple and Flexible In Vitro Three-dimensional Invasion Assay Protocol

Moving Upwards: A Simple and Flexible In Vitro Three-dimensional Invasion Assay Protocol

向上移动: 一个简单而灵活的体外三维入侵检测协议

Although 3D invasion assays have been developed, the challenge remains to study cells without affecting the integrity of their microenvironment. ...

Orthotopic Transplantation of Syngeneic Lung Adenocarcinoma Cells to Study PD-L1 Expression

合成肺腺癌细胞原位移植研究 pd-l1 表达

The use of mouse models is indispensable for studying the pathophysiology of various diseases. With respect to lung cancer, several models are available, ...

Mesenchymal Stem Cell Isolation from Pulp Tissue and Co-Culture with Cancer Cells to Study Their Interactions

骨髓间充质干细胞在浆组织中的分离及与癌细胞联合培养研究其相互作用

Cancer as a multistep process and complicated disease is not only regulated by individual cell proliferation and growth but also controlled by tumor ...

Establishment and Characterization of Three Afatinib-resistant Lung Adenocarcinoma PC-9 Cell Lines Developed with Increasing Doses of Afatinib

建立和表征三个抗阿法替尼的肺腺癌PC-9细胞系与增加剂量的阿法替尼开发

Acquired resistance to molecular target inhibitors is a severe problem in cancer therapy. Lung cancer remains the leading cause of cancer-related death in ...

In Vitro Assay to Study Tumor-macrophage Interaction

体外测定研究肿瘤-巨噬菌体相互作用

Tumor associated macrophages (TAMs) account for a large percentage of cells in the tumor mass for different types of cancers. Glioblastoma (GBM), a ...

In Vitro Evaluation of Oncogenic Transformation in Human Mammary Epithelial Cells

人类乳腺上皮细胞致癌转化的体外评价

Tumorigenesis is a multi-step process in which cells acquire capabilities&#160;that allow their growth, survival, and dissemination under hostile ...

Using Computer-based Image Analysis to Improve Quantification of Lung Metastasis in the 4T1 Breast Cancer Model

利用计算机图像分析提高4T1乳腺癌模型中肺转移的定量

Breast cancer is a devastating malignancy, accounting for 40,000 female deaths and 30% of new female cancer diagnoses in the United States in 2019 alone. ...

Chemical Conjugation of a Purified DEC-205-Directed Antibody with Full-Length Protein for Targeting Mouse Dendritic Cells In Vitro and In Vivo

Chemical Conjugation of a Purified DEC-205-Directed Antibody with Full-Length Protein for Targeting Mouse Dendritic Cells In Vitro and In Vivo

纯化的DEC-205定向抗体与全长蛋白的化学偶联，用于在体外和体内靶向小鼠树突状细胞

Targeted antigen delivery to cross-presenting dendritic cells (DC) in vivo efficiently induces T effector cell responses and displays a valuable approach ...