Ezrin protein’s role on cell migration in gastric cancer and identification of m6a-modified sites

Resumo

Background: Gastric cancer (GC) is the fourth leading cause of cancer-related deaths, often diagnosed in late stages of the disease, limiting treatment options [1-2]. The absence of new therapies and proper diagnosis calls for an exploration of different potential targets. Epitranscriptomics studies RNA chemical modifications such as N6-methyladenosine (m6A), lately studied to understand its effect on cancer progression and potential treatment [3-4]. YTHDF3 is an m6A reader and its role in migration was established by the “Differentiation and Cancer” group at i3S (unpublished data, manuscript submitted). In this study we focused on EZRIN (EZR), which is a target of YTHDF3. Aims: To inhibit EZR expression using NSC668384, study the impact on migration/metastization and on Paclitaxel response and identify the m6A methylation sites on the EZR transcript using the SELECT method. Methods: We inhibited EZR expression with NSC668384 in two GC cell lines, characterizing the effect on cell viability, cell migration through a migration assay, and performed a combined treatment of NSC668384 with Paclitaxel. We also used the SELECT method to identify m6A- modified sites in the EZR transcript. Results: EZR is involved in cell migration and its downregulation sensitizes cells to Paclitaxel treatment. We successfully detected two methylated sites in the EZR transcript. Conclusions: The loss of migration capacity upon EZR inhibition aligns with previous studies showing EZR's critical role in cancer cell motility [5-9]. The increased sensitivity of SNU638 cells to Paclitaxel supports findings from other malignancies where EZR inhibition enhances chemotherapy efficacy, indicating a shared mechanism across cancers [10]. Additionally, the presence of m6A modifications in the EZR transcript highlights the need to investigate their functional impact on EZR regulation and therapeutic responses. EZR’s inhibition has therapeutical value, and the identification of m6A-sites will allow research on these modifications’ role which may result in development of new treatment strategies.