Generation and Characterisation of Cisplatin-Resistant Non-Small Cell Lung Cancer Cell Lines Displaying a Stem-Like Signature

Abstract

IntroductionInherent 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.MethodsAn 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.ResultsCharacterisation 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.ConclusionOur 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.

ABCG2 regulates the pattern of self-renewing divisions in cisplatin-resistant non-small cell lung cancer cell lines

“…Loss-of-function mutations of cell polarity and cell fate determinants induce neuroblasts to divide symmetrically, leading to an increase in number, tissue overgrowth and ultimately transplantable tumors that resemble mammalian cancers (9). Recent evidence has revealed that cisplatin-resistant subpopulations of NSCLC cells have a putative stem-like character, including increased invasive ability and tumorigenic ability (25). Thus, our findings are consistent with previous studies, which showed an increased proportion of symmetric division in cisplatin-resistant NSCLC cells compared to the parental cells.…”

HIPEC ROC I: A phase i study of cisplatin administered as hyperthermic intraoperative intraperitoneal chemoperfusion followed by postoperative intravenous platinum‐based chemotherapy in patients with platinum‐sensitive recurrent epithelial ovarian cancer

“…Intratumoural heterogeneity of cisplatininduced DNA adducts may identify subpopulations of ovarian cancer cells with a putative stem-like signature as shown in non-small cell lung cancer cell lines. 37 Inherent limitations of this phase I study include the small sample size with lack of quality-of-life data and a lack of cost effectiveness analysis, which will play an important role in the future development of this multimodality treatment. The median operative time of almost 8 h reflects the complexity of this combined procedure.…”

MicroRNA-100 Resensitizes Resistant Chondrosarcoma Cells to Cisplatin through Direct Targeting of mTOR

“…Zhe Zhu 1,2& , Cun-Ping Wang 2& , Yin-Feng Zhang 3 , Lin Nie 1 * rate of initial responses, cisplatin resistance limits its utilizations in cancer patients because some cancer cells develop acquired cisplatin resistance eventually (Negoro et al, 2007;Galluzzi et al, 2012;Barr et al, 2013). The mechanisms of cisplatin resistance are not fully understood yet.…”