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Trejo Vazquez, Jessica Angelina

University of British Columbia

Lung cancer is the leading cause of cancer deaths worldwide, largely due to metastasis. Its main subtype is lung adenocarcinoma (LUAD). Communication between cancer cells and cells within the tumor microenvironment, such as fibroblasts, can promote metastasis. This interaction can be mediated by extracellular vesicles (EVs) transferring bioactive cargo. Thus, this study aimed to identify the impact of EVs derived from LAC cells on lung fibroblasts. For EV isolation, conditioned media derived from H2073 and H1437 LAC cell lines were ultracentrifuged multiple times after debris removal. Nanoparticle Tracking Analysis was used for quantification and size characterization and EV-associated markers were assessed by western blot (WB). For lung fibroblast activation, H6013 primary adult cells were treated with 1xPBS, TGFβ (10 ng/μL) or EVs (7.13x10¹¹) every 24 hours over 3 days. Protein (for WB) and RNA (for gene expression profiling) were harvested 48 hours after treatment. RNA was extracted using the Qiagen miRNeasy kit and sequenced by Illumina NextSeq2000. Most vesicles produced by both cell lines were between 50 and 250 nm in diameter. Fibroblasts treated with TGFβ or EVs had an increase in CAF markers (mainly α-SMA), especially in the EV group compared to the PBS treatment group. The activated fibroblasts shared differentially expressed genes in the TGFβ and EV treatments; however, there was an increase in some of the genes associated with epithelial-mesenchymal transition and cell cycle only in the EV-treated group, indicating unique features of CAF activation and the same was seen for the pathway analysis. Therefore, LUAD EVs have a role in CAF activation by using both similar and unique pathways to TGFβ activation. Further understanding of the genes distinct to the EV-treated fibroblasts could lead to the identification of novel genes involved in CAF activation, and thus aid in the development of novel therapeutics that could improve patient outcomes.

Text

2024-05-16

Attribution-NonCommercial-NoDerivatives 4.0 International

http://hdl.handle.net/2429/88329

Interdisciplinary Oncology

University of British Columbia

UBCV

http://creativecommons.org/licenses/by-nc-nd/4.0/