Anti-Tumor Effects of Biomimetic Sulfated Glycosaminoglycans on Lung Adenocarcinoma Cells in 2D and 3D In Vitro Models.

Nada Al Matari,Ansam Sinjab,Humam Kadara,Rami Mhanna,Wassim Abou-Kheir,Hiba Mshiek,George Deeb

doi:10.3390/molecules25112595

Abstract

Lung cancer development relies on cell proliferation and migration, which in turn requires interaction with extracellular matrix (ECM) components such as glycosaminoglycans (GAGs). The mechanisms through which GAGs regulate cancer cell functions are not fully understood but they are, in part, mediated by controlled interactions with cytokines and growth factors (GFs). In order to mechanistically understand the effect of the degree of sulfation (DS) of GAGs on lung adenocarcinoma (LUAD) cells, we synthesized sulfated alginate (AlgSulf) as sulfated GAG mimics with DS = 0.0, 0.8, 2.0, and 2.7. Human (H1792) and mouse (MDA-F471) LUAD cell lines were treated with AlgSulf of various DSs at two concentrations 10 and 100 µg/mL and their anti-tumor properties were assessed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), trypan blue exclusion, and wound healing assays for 2D models and sphere formation assay for the 3D model. The proliferation and number of live MDA-F471 cells at the concentration of 100 µg/mL decreased significantly with the increase in the DS of biomimetic GAGs. In addition, the increase in the DS of biomimetic GAGs decreased cell migration (p < 0.001 for DS = 2.0 and 2.7 compared to control) and decreased the diameter and number of spheres formed (p < 0.001). The increased DS of biomimetic GAGs attenuated the expression of cancer stem cell (CSC)/progenitor markers in the 3D cultures. In conclusion, GAG-mimetic AlgSulf with increased DS exhibit enhanced anti-proliferative and migratory properties while also reducing growth of KRAS-mutant LUAD spheres in vitro. We suggest that these anti-tumor effects by GAG-mimetic AlgSulf are possibly due to differential binding to GFs and consequential decreased cell stemness. AlgSulf may be suitable for applications in cancer therapy after further in vivo validation.

Highlights

Cancer is the leading cause of death worldwide with approximately 9.6 million cancer-related deaths in 2018 [1]
lung adenocarcinomas (LUAD) are thought to be mutually exclusively driven by epithelial growth factor receptor (EGFR)-a dependent signaling and Kirsten rat sarcoma viral oncogene homolog (KRAS) pathway [6]
We showed using a wound healing/ scratch assay that the migration of 2D cultured KRAS-mutant human and murine LUAD cells decreased when cells were exposed to biomimetic sulfated GAGs

Summary

Introduction

Cancer is the leading cause of death worldwide with approximately 9.6 million cancer-related deaths in 2018 [1]. 19% of all newly diagnosed cancers), and the leading cause of cancer deaths (19% of cancer deaths) [2,3]. Molecules 2020, 25, 2595 cancer and non-small cell lung cancer (NSCLC), contributing to about 15% and 85% of all lung cancer incidences, respectively [4,5]. NSCLC can be further histologically categorized into squamous-cell carcinomas, lung adenocarcinomas (LUAD), and large-cell lung carcinomas. Somatic activating mutations in KRAS are the most prevalent oncogenic driver mutations in LUAD accounting for 25–30% of all mutations and are tightly associated with smoking [6,7].

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