Multidrug resistance characterization in multicellular tumour spheroids from two human lung cancer cell lines

Abstract

BackgroundMost of the knowledge about the mechanisms of multidrug resistance in lung cancer has been achieved through the use of cell lines isolated from tumours cultivated either in suspensions of isolated cells or in monolayers and following exposition to different cytostatic agents. However, tumour cell lines growing as multicellular tumour spheroids (MTS) frequently develop multicellular resistance in a drug-independent form. The aim of this study was to characterize the phenotypic and functional differences between two human NSCLC cell lines (INER-37 and INER-51) grown as traditional monolayer cultures versus as MTS.MethodsAfter 72 hours treatment with anticancer drugs, chemosensitivity in monolayers and tumour spheroids cultures was assessed using MTT assay. Reverse transcription-polymerase chain reaction was employed to detect the mRNAs of multidrug resistance-related genes. The expression of P-gp was analyzed by immunohistochemical staining and cell cycle profiles were analyzed using FACS.ResultsThe results indicate that when grown as MTS each lung cancer cell line had different morphologies as well as and abrogation of cell proliferation with decrease of the G2/M phase. Also, MTS acquired multicellular resistance to several chemotherapeutic agents in only a few days of culture which were accomplished by significant changes in the expression of MDR-related genes.ConclusionOverall, the MTS culture changed the cellular response to drugs nevertheless each of the cell lines studied seems to implement different mechanisms to acquire multicellular resistance.

3D tumor spheroids as in vitro models to mimic in vivo human solid tumors resistance to therapeutic drugs

“…Despite that, these cells are able to express cytokines, chemokines and growth factors involved in tumorigenesis and therapeutics resistance profile (reviewed by M. ). Barrera- Rodríguez and Fuentes (2015) suggested that INER-37 and INER-51 lung cancer spheroids are more resistance toward etoposide, teniposide, doxorubicin, or camptothecin when compared with 2D cell cultures, a result that was associated to the quiescent cell subpopulation found in the spheroids. In a different study, Gong et al (2015) analyzed the cell cycle of MCF-7 breast cancer cells cultured in monolayer or spheroids through flow cytometry.…”

A straightforward method to produce decellularized dermis-based matrices for tumour cell cultures

“…In respect to conventional 2D cultures, the added dimensionality enables the formation of large and variable tumour spheroids similar in morphology to those observed in vitro . Additionally, spheroids in 3D displayed a better resistance to drug treatments (Barrera‐Rodriguez and Morales Fuentes, ). Synthetic scaffolds can be produced with predefined shape, porosity and stiffness.…”

Chemoresistance of Lung Cancer Cells: 2D and 3D In Vitro Models for Anticancer Drug Screening

“…Activations of mechanisms related to drug efflux in spheroid can be responsible for increased resistance to the drugs. In 2015, Rodriguez et al showed an increased expression of MDR-1 and P-glycoprotein in spheroids of INER-37 human NSCLC cell line compared to 2D culture likely responsible for increased drug resistance [58]. Several studies have indicated CSC enrichment in spheroids as possible explanation for drug resistance and metastasis [59,60].…”