HIV-1 latency reactivation by VP-16
10.3760/cma.j.cn112866-20210803-00137
- VernacularTitle:VP-16对HIV-1潜伏感染再激活作用的研究
- Author:
Cancan CHEN
1
;
Weiqi ZHANG
;
Hanyu MA
;
Ying TUO
Author Information
1. 中山大学附属第一医院病理科，广州 510080
- Keywords:
VP-16;
HIV-1;
Latent infection;
SIRT1
- From:
Chinese Journal of Experimental and Clinical Virology
2022;36(1):8-14
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To explore the effect of the etoposide (VP-16) on the reactivation of human immunodeficiency virus (HIV-1) from latent infection and study its potential molecular mechanism.Methods:The HIV-1 latently infected cell line J-Lat was treated with DMSO, VP-16 and vorinostat (SAHA), flow cytometry was used to detect the positive ratio of green fluorescent protein (GFP) in J-Lat, which can reflect the efficacy of high concentration VP-16 on reactivation. Then the reactivation efficiency of VP-16 on HIV-1 latently infected cells at different concentration and treatment time was measured by flow cytometry. The expression of CD25, CD69 and interleukin-6 (IL-6) of the VP-16-treated J-Lat cells were also detected in the same way. The effect of VP-16 on the transcription of long terminal repeat (LTR) was tested using a dual-luciferase reporter assay. The protein expression of the silent information regulator 1 (SIRT1) and the acetylated nuclear factor κB (NF-κB) p65 under the effect of VP-16 was checked by western blot (WB). Lentivirus-mediated SIRT1 short-hairpin RNA (SIRT1-shRNA) was employed to knock down SIRT1, and then tested for efficiency of reactivation.Results:The results of flow cytometry showed that VP-16 specifically reactivated HIV-1 latently infected J-Lat cells in a concentration and time-dependent manner. Meanwhile, the expression of CD25 and CD69 was upregulated to a certain extent, but the expression of IL-6 was not affected. Dual-luciferase reporter assay suggested that VP-16 positively regulated the transcription of HIV-1 LTR through NF-κB signaling pathway. WB results indicated that VP-16 can inhibit the protein expression of SIRT1 and promote the acetylation of NF-κB p65. Lentivirus-mediated SIRT1-shRNA successfully knocked down the mRNA and protein expression of SIRT1, and reactivated the HIV-1 latency effectively.Conclusions:VP-16 upregulates the acetylation of NF-κB p65 by inhibiting the expression of SIRT1, which subsequently promotes the transcription of HIV-1 LTR and reactivates the latent HIV-1 infection.
