Selinexor, a selective inhibitor of nuclear export, shows anti-proliferative and anti-migratory effects on male germ cells in vitro

Abstract Background Selinexor (SLX), a selective inhibitor of nuclear export (SINE), has been shown to interfere with nuclear export mechanisms and to exert antitumor effects in a variety of cancer cell types. It is known to regulate multiple fundamental cellular processes, including the DNA damage response, cell proliferation, and stress signaling pathways. Nevertheless, its potential effects on reproductive cells remain inadequately characterized. The present study was aimed to investigate the cytotoxic, apoptotic, anti-proliferative and anti-migratory effects of SLX on GC1 (spermatogonia) and GC2 (spermatid) cell lines, alongside its influence on DNA damage and oxidative stress. Methods Cytotoxicity was assessed using the MTT assay. Cell proliferation capacity was evaluated via colony formation assay, while cell migration was analyzed using in vitro wound healing model. Apoptosis, oxidative stress, and DNA damage were investigated using immunocytochemical analyses of Cas-3, Bax, iNOS, ATM, and BRCA1 proteins. Additionally, Annexin V-FITC/PI staining was performed to detect the apoptotic cell population by flow cytometry. Results SLX treatment led to concentration- and time dependent cytotoxicity and colony formation assay revealed a marked reduction in proliferative capacity, in both cell lines. Wound healing analyses demonstrated that SLX effectively suppressed cell migration. Flow cytometry analysis showed that the live cell population decreased, whereas the late apoptotic cell population increased. Additionally, it was observed that Cas-3 and Bax immunoreactivities increased in the SLX groups compared to the control groups. Moreover, a significant increase in the immunoreactivity of ATM, BRCA1 and iNOS proteins, which are key indicators of DNA damage and oxidative stress, was observed. Conclusion The data suggest that SLX may decrease cell viability, induce apoptosis, inhibit cell migration and increase DNA damage and cellular stress in male germ cells. Given these effects, SLX should be carefully examined for its potential reproductive toxicity. Further studies are warranted to explore its long-term impact on male fertility.