A preliminary study on thapsigargin promoting coxsackievirus B3 replication through activating transcription factor 6 pathway

Abstract

Objective To investigate the effect of thapsigargin (TG) which can induce endoplasmic reticulum stress (ERS) on the replication of coxsackievirus B 3 (CV-B3). Methods After 10 MOI CV-B3 infected HeLa cells were exposed 0.25 μmol/L TG for 3 h, 6 h and 9 h, virus RNA of HeLa cells were extracted and viral replication was evaluated by real time PCR. After 0.25 μmol/L、0.08 μmol/L and 0.025 μmol/L TG exposed, the plaque of CV-B3 was used to confirm further replication of CV-B3. To verify TG induced ERS through three signal pathway, one of among PERK, ATF6 and IRE1 inhibitors GSK2656157, AEBSF and STF-083010, and 0.25 μmol/L TG were used in HeLa cells infected with 10 MOI CV-B3, replication of CV-B3 was evaluated by qRT-PCR. Results The stimulation of TG did not induce increase of virus replication after post-infection 3 h. However, TG induced replication of virus to increase 2.5 times after post-infection 6 h and 158.6 times after post-infection 9 h. And, the area of viral plaque was significantly increased. ATF6 inhibitors AEBSF significantly inhibited promotion of virus replication from TG. Conclusions TG can promote the replication of CV-B3 through ATF6 signal pathway. Key words: Coxsachieviuus B3; Thapsigargin; Endoplasmic reticulum stress; Plaque; ATF6 pathway