Generation and characterisation of cisplatin-resistant non-small cell lung cancer cell lines displaying a stem-like signature.
Generation and characterisation of cisplatin-resistant non-small cell lung cancer cell lines displaying a stem-like signature.
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DOI：
10.1371/journal.pone.0054193
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
O'Byrne KJ
中科院分区：
文献类型：
--
作者：
Barr MP;Gray SG;Hoffmann AC;Hilger RA;Thomale J;O'Flaherty JD;Fennell DA;Richard D;O'Leary JJ;O'Byrne KJ
固有和获得性顺铂耐药性降低了该药物在非小细胞肺癌（NSCLC）治疗中的有效性。了解这一过程背后的分子机制可能会促使新型药物的研发，以提高顺铂的敏感性。
在一组非小细胞肺癌细胞系（A549、SKMES - 1、MOR、H460）中建立了顺铂耐药的同基因模型。在12个月的时间里，从原始的、年龄匹配的亲本细胞（PT）中获得了顺铂耐药（CisR）细胞系，并随后对其进行了特征分析。在亲本细胞和顺铂耐药细胞之间进行了增殖（MTT）和克隆形成存活测定（结晶紫）。通过流式细胞术分析检测了细胞对顺铂诱导的细胞凋亡的反应以及细胞周期分布。除了上皮 - 间质转化（EMT）蛋白、c - Met和β - 连环蛋白外，还检测了一组癌症干细胞和多能性标志物。还评估了顺铂 - DNA加合物的形成、DNA损伤（γH2AX）以及细胞对铂的摄取（电感耦合等离子体质谱法，ICP - MS）。
特征研究表明，肺肿瘤细胞对顺铂的增殖能力下降，对顺铂诱导的细胞死亡的抵抗力增强，耐药细胞在细胞周期的G0/G1期积累，克隆形成存活能力提高。此外，耐药细胞显示出一种假定的干细胞样特征，与相应的亲本细胞相比，CD133⁺/CD44⁺细胞表达增加，乙醛脱氢酶（ALDH）活性增强。干细胞标志物Nanog、Oct - 4和SOX - 2以及上皮 - 间质转化标志物c - Met和β - 连环蛋白均显著上调。虽然耐药亚系在治疗时对顺铂的摄取减少，但与亲本细胞系相比，观察到顺铂 - GpG DNA加合物形成减少，γH2AX焦点显著减少。
我们的研究结果确定了具有假定干细胞样特征的非小细胞肺癌细胞的顺铂耐药亚群，进一步了解了与肺癌顺铂耐药表型相关的细胞事件。
Inherent and acquired cisplatin resistance reduces the effectiveness of this agent in the management of non-small cell lung cancer (NSCLC). Understanding the molecular mechanisms underlying this process may result in the development of novel agents to enhance the sensitivity of cisplatin. An isogenic model of cisplatin resistance was generated in a panel of NSCLC cell lines (A549, SKMES-1, MOR, H460). Over a period of twelve months, cisplatin resistant (CisR) cell lines were derived from original, age-matched parent cells (PT) and subsequently characterized. Proliferation (MTT) and clonogenic survival assays (crystal violet) were carried out between PT and CisR cells. Cellular response to cisplatin-induced apoptosis and cell cycle distribution were examined by FACS analysis. A panel of cancer stem cell and pluripotent markers was examined in addition to the EMT proteins, c-Met and β-catenin. Cisplatin-DNA adduct formation, DNA damage (γH2AX) and cellular platinum uptake (ICP-MS) was also assessed. Characterisation studies demonstrated a decreased proliferative capacity of lung tumour cells in response to cisplatin, increased resistance to cisplatin-induced cell death, accumulation of resistant cells in the G0/G1 phase of the cell cycle and enhanced clonogenic survival ability. Moreover, resistant cells displayed a putative stem-like signature with increased expression of CD133+/CD44+cells and increased ALDH activity relative to their corresponding parental cells. The stem cell markers, Nanog, Oct-4 and SOX-2, were significantly upregulated as were the EMT markers, c-Met and β-catenin. While resistant sublines demonstrated decreased uptake of cisplatin in response to treatment, reduced cisplatin-GpG DNA adduct formation and significantly decreased γH2AX foci were observed compared to parental cell lines. Our results identified cisplatin resistant subpopulations of NSCLC cells with a putative stem-like signature, providing a further understanding of the cellular events associated with the cisplatin resistance phenotype in lung cancer.
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影响因子：
12.4
作者：
Eramo, A.;Lotti, F.;De Maria, R.
通讯作者：
De Maria, R.
影响因子：
7.2
作者：
Crawford, Nyree;Chacko, Alex D.;Fennell, Dean A.
通讯作者：
Fennell, Dean A.
影响因子：
6.5
作者：
Liedert, B;Materna, V;Lage, H
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Lage, H
表达 CD44 的非小细胞肺癌细胞富含干细胞样特性。
影响因子：
3.7
作者：
Leung, Elaine Lai-Han;Fiscus, Ronald R.;Tung, James W.;Tin, Vicky Pui-Chi;Cheng, Lik Cheung;Sihoe, Alan Dart-Loon;Fink, Louis M.;Ma, Yupo;Wong, Maria Pik
通讯作者：
Wong, Maria Pik
影响因子：
3.7
作者：
Chen, Yu-Chih;Hsu, Han-Shui;Chen, Yi-Wei;Tsai, Tung-Hu;How, Chorng-Kuang;Wang, Chien-Ying;Hung, Shih-Chieh;Chang, Yuh-Lih;Tsai, Ming-Long;Lee, Yi-Yen;Ku, Hung-Hai;Chiou, Shih-Hwa
通讯作者：
Chiou, Shih-Hwa