Anti-HTLV-1 Virus Transmission and Inhibition of Cell Proliferation of Adult T-Cell Leukemia by Ritona-vir and the Mechanism Study
OBJECTIVE To observe the inhibitory effect of ritonavir on human T-cell leukemia virus type-1(HTLV-1)transmis-sion and malignant proliferation of adult T-cell leukemia(ATL)cells,and explore its molecular mechanism.METHODS The prolif-eration and vitality of ritonavir on various leukemic cells were evaluated by CCK-8 and colony formation assay.The effects of ritonavir on HTLV-1 virus transmission were detected by flow cytometry,dual luciferase reporter gene technique,qPCR and Western blot.The effects of ritonavir on the cell cycle and apoptosis of ATL cells were examined through flow cytometry.RESULTS Ritonavir could inhibit the proliferation of four ATL cell lines and the clonal proliferation of HTLV-1 positive cell lines.The former exhibited a signifi-cant dose-effect relationship and had a more pronounced inhibitory effect on HTLV-1 positive cell lines(P＜0.05).Additionally,the administration of ritonavir immediately after co-culture of HTLV-1 positive cell lines with JETWT35 cells resulted in a significant down-regulation of red fluorescent protein expression in JETWT35 and inhibited the transmission of HTLV-1 virus into recipient cells(P＜0.01).Upon immediate addition of ritonavir to the co-culture system of HTLV-1 positive cell lines and Jurkat cells,there was a notable inhibition of HTLV-1-related gene Tax and other genes mRNA in recipient cells(P＜0.01);however,no significant effect was observed when ritonavir was added 12 h after virus transmission.Morever,ritonavir demonstrated a does-dependent inhibition of gp46 expression on the cell membrane of the HTLV-1 positive cell line ATL-T,thereby suppressing the production of HTLV-1 virus(P＜0.01).Ritonavir impeded cell progression into G1 phase and facilitated apoptosis,with the apoptosis rate of HTLV-1 positive cell lines being significantly greater than that of HTLV-1 negative cell lines(P＜0.05 or P＜0.01).CONCLUSION Ritonavir exerts inhibitory effects on the production and transmission of HTLV-1 virus by diminishing the activity of WT-Luc virus promoter,suppressing the expression of HTLV-1-related virus genes(Tax,HBZ,Gag,Pol,and Env).Additionally,it inhibits the expression of the HTLV-1-positive membrane surface envelope protein subunit gp46.Futhermore,ritonavir induces apoptosis in ATL cells by arresting cell cycle in the G1 phase,thereby effectively suppressing cell proliferation.
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