Establishment of Acquired Cisplatin Resistance in Ovarian Cancer Cell Lines Characterized by Enriched Metastatic Properties with Increased Twist Expression

Abstract

Ovarian cancer (OC) is the most lethal of the gynecologic cancers, and platinum-based treatment is a part of the standard first-line chemotherapy regimen. However, rapid development of acquired cisplatin resistance remains the main cause of treatment failure, and the underlying mechanism of resistance in OC treatment remains poorly understood. Faced with this problem, our aim in this study was to generate cisplatin-resistant (CisR) OC cell models in vitro and investigate the role of epithelial–mesenchymal transition (EMT) transcription factor Twist on acquired cisplatin resistance in OC cell models. To achieve this aim, OC cell lines OV-90 and SKOV-3 were exposed to cisplatin using pulse dosing and stepwise dose escalation methods for a duration of eight months, and a total of four CisR sublines were generated, two for each cell line. The acquired cisplatin resistance was confirmed by determination of 50% inhibitory concentration (IC50) and clonogenic survival assay. Furthermore, the CisR cells were studied to assess their respective characteristics of metastasis, EMT phenotype, DNA repair and endoplasmic reticulum stress-mediated cell death. We found the IC50 of CisR cells to cisplatin was 3–5 times higher than parental cells. The expression of Twist and metastatic ability of CisR cells were significantly greater than those of sensitive cells. The CisR cells displayed an EMT phenotype with decreased epithelial cell marker E-cadherin and increased mesenchymal proteins N-cadherin and vimentin. We observed that CisR cells showed significantly higher expression of DNA repair proteins, X-ray repair cross-complementing protein 1 (XRCC1) and poly (ADP-ribose) polymerases 1 (PARP1), with significantly reduced endoplasmic reticulum (ER) stress-mediated cell death. Moreover, Twist knockdown reduced metastatic ability of CisR cells by suppressing EMT, DNA repair and inducing ER stress-induced cell death. In conclusion, we highlighted the utilization of an acquired cisplatin resistance model to identify the potential role of Twist as a therapeutic target to reverse acquired cisplatin resistance in OC.

Interplay Between Thiamine and p53/p21 Axes Affects Antiproliferative Action of Cisplatin in Lung Adenocarcinoma Cells by Changing Metabolism of 2-Oxoglutarate/Glutamate

“…Remarkably, the shape of attached A549 p21– cells demonstrate elongation absent in the A549 WT cells ( Figure 1G ). The morphological change is reminiscent of epithelial-mesenchymal transition, known to reduce cisplatin sensitivity ( Deng et al, 2019 ; Bahar et al, 2020 ; Brum et al, 2020 ; Chen et al, 2020 ; Siraj et al, 2020 ), which is also observed in our experiments ( Figure 1E ).…”

“…Permanent endoplasmic reticulum stress has been shown to contribute to mechanisms of the cisplatin resistance ( Bahar et al, 2020 ; Chen et al, 2020 ). In neurons, such stress is known to be induced by thiamine deficiency ( Wang et al, 2007 ; Liu et al, 2017 ; Xu et al, 2019b ).…”

Combination of Niraparib, Cisplatin and Twist Knockdown in Cisplatin-Resistant Ovarian Cancer Cells Potentially Enhances Synthetic Lethality through ER-Stress Mediated Mitochondrial Apoptosis Pathway

“…The cells were cultured in their respective ATCC recommended growth supplemented with fetal bovine serum (FBS). Two CisR OC cells, OV90/CisR and SKOV3/CisR were generated by the stepwise increment of cisplatin dosing method described in our previous publication [ 27 ].…”

“…To generate in vitro three–dimensional (3D) spheroidal model utilized combination of hanging drop and hydrogel scaffolds modified methods [ 27 , 84 ]. The details of generation of three dimensional (3D) spheroidal model are described in the supporting information, Supplementary materials and methods, Section S1.1 .…”

“…Twist is a member of the core transcription factor helix-loop-helix (bHLH), known to be an epithelial–mesenchymal transition (EMT) master regulator associated with tumor recurrence and chemo-resistance [ 19 , 20 , 21 , 22 , 23 , 24 , 25 , 26 ]. Functionally, Twist was identified as a potential oncogene and our previous studies have identified that Twist also contributed to acquire cisplatin resistance in OC cells [ 27 , 28 , 29 ]. In several studies, Twist has been linked to the development of resistance to platinum-based chemotherapy and metastasis in cancer [ 20 , 30 , 31 , 32 , 33 , 34 , 35 ].…”