Establishment and Characterization of a Topotecan Resistant Non-small Cell Lung Cancer NCI-H460/TPT10 Cell Line

Chao-Yun Cai,Ying-Fang Fan,Dong-Hua Yang,Qiu-Xu Teng,Wei Zhang,Jing-Quan Wang,Kimberly W. Lu,John N. D. Wurpel,Zi-Ning Lei,Zhe-Sheng Chen

doi:10.3389/fcell.2020.607275

Abstract

While topotecan (TPT) is a first- and second-line chemotherapeutic drug in treating lung cancer, the development of drug resistance in tumors still reserves as a major obstacle to chemotherapeutic success. Therefore, a better understanding of the mechanisms of topotecan resistance is critical. In this study, the first topotecan-resistant human non-small cell lung cancer (NSCLC) cell line, termed NCI-H460/TPT10, was established from the parental NCI-H460 cell line. NCI-H460/TPT10 cells exhibited a 394.7-fold resistance to TPT, and cross-resistance to SN-38, mitoxantrone, and doxorubicin, compared to parental NCI-H460 cells. Overexpression of ABCG2 localized on the cell membrane, but not ABCB1 or ABCC1, was found in NCI-H460/TPT10 cells, indicating that ABCG2 was likely to be involved in topotecan-resistance. This was confirmed by the abolishment of drug resistance in NCI-H460/TPT10 cells after ABCG2 knockout. Moreover, the involvement of functional ABCG2 as a drug efflux pump conferring multidrug resistance (MDR) was indicated by low intracellular accumulation of TPT in NCI-H460/TPT10 cells, and the reversal effects by ABCG2 inhibitor Ko143. The NCI-H460/TPT10 cell line and its parental cell line can be useful for drug screening and developing targeted strategies to overcome ABCG2-mediated MDR in NSCLC.

Highlights

Lung cancer is the most frequently diagnosed cancer worldwide and is one of the most common causes of cancer-related death (Bray et al, 2018)
The topotecan-resistant nonsmall cell lung cancer (NSCLC) cell line NCI-H460/TPT10 was eventually developed by selecting the parental NCI-H460 cells in stepwise increasing concentrations of topotecan until cells survive in topotecan at the concentration up to 10 μM
After a 5-month of selection with topotecan and 3 months culturing without topotecan, NCI-H460/TPT10 cells conferred a 394.7-fold resistance against topotecan compared to parental NCI-H460 cells, confirming the acquisition of topotecan resistance in the newly established cell line

Summary

Introduction

Lung cancer is the most frequently diagnosed cancer worldwide and is one of the most common causes of cancer-related death (Bray et al, 2018). Topotecan Resistant Lung Cancer Cell-Line treatment is initially effective, conventional chemotherapies often fail due to the acquired and/or intrinsic multidrug resistance (MDR) in tumors, leading to a high risk of local-regional recurrence and distant relapse (Wangari-Talbot and HopperBorge, 2013; Iglesias et al, 2018). Many mechanisms can be associated with MDR in lung cancers, such as decreased uptake of drugs that enter cells via transporters, increased efflux of drugs from cells, and changes in the cellular response to drugs, including decreased cell apoptosis, altered drug metabolism, mutated drug target, and increased DNA repair (Wangari-Talbot and Hopper-Borge, 2013; Iglesias et al, 2018; Terlizzi et al, 2019). MDR can be multifactorial, one of the most widely observed mechanisms has been the increased drug efflux linked to the overexpression of transmembrane ATP-binding cassette (ABC) transporters (Robey et al, 2018)

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