VCOM Louisiana Research Day Program Book 2024

Biomedical Research: Section 2

Archana Shrestha, BPharm 1 ; Prajesh Shrestha, BPharm 1 ; Md Anamul Haque, MPharm 1 ; Md Mahbubul Hoque, MPharm 1 ; Margarita Lamprou, PhD 2 ; Constantinos M. Mikelis, PhD 2 ; George Mattheolabakis, PhD 1 1 School of Basic Pharmaceutical and Toxicological Sciences, College of Pharmacy, University of Louisiana Monroe; 2 Laboratory of Molecular Pharmacology, Department of Pharmacy, University of Patras 17 LENTIVIRUS-MEDIATED STABLE DUAL MIR-143 AND MIR-506 UPREGULATION DEMONSTRATES IMPROVED RESPONSE AGAINST A549 CELL PROLIFERATION, MIGRATION, SURVIVAL AND TUMOR GROWTH PROGRESSION COMPARED TO INDIVIDUAL MIR DEREGULATION

Background: In the United States, non-small cell lung cancer (NSCLC) is the most frequently occurring lung cancer type that accounts for >80% of all lung cancer cases with a 5-year survival rate of only 28%. In recent years, individual micro-RNA (miR) therapies for NSCLC have been widely explored. However, several miRs are deregulated at a time in several cancer types, so emphasis must be put on evaluating miR combinatory effects against cancer cell progression. Objective: In our study, we previously identified two miRs, miR-143 and miR-506, that were significantly downregulated in A549 cells. We developed lentiviral-mediated stable transduction in A549 cells to either up- or downregulate these miRs in combination or individually to evaluate various cellular responses compared to the A549 miR-GFP control stable transfected cells. Methods: After the initial characterization of the respective A549 miR-stable cell groups using Taqman qPCR and fluorescence-activated cell sorting (FACS), we evaluated the effects of miR deregulation on cell proliferation, cell cycle progression, cell migration and motility, colony formation, and tumor growth progression.

Results: Compared to the individual miR deregulations, the combined miR upregulation demonstrated a significant increase in the cell doubling time in contrast to its respective downregulation. Using wound healing and transwell migration assays, we observed that the individual miR upregulations did not indicate a significant outcome as compared to the miR-GFP control and their respective miR-downregulations. Meanwhile, the combined miR-143/506 upregulation showed a decreased cell migration compared to both A549 miR-GFP-control and combined miR downregulation. Furthermore, we performed subcutaneous injection of all A549 stable miR-deregulated groups in mice, where we observed that dual upregulation of miR-143/506 resulted in significantly diminished tumor growth contrary to the increased tumor growth in dual downregulation of both miRs. Conclusions: Therefore, these results indicate a well-defined, miR-dependent outcome of combined up- and downregulation of miR 143 and miR-506, which leads to an improved understanding of A549 cancer progression. Thus, these findings present dual deregulation of miRs as a promising strategy for lung cancer therapy.

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