Journal of Jilin University(Medicine Edition) ›› 2021, Vol. 47 ›› Issue (4): 874-881.doi: 10.13481/j.1671-587X.20210408

• Research in basic medicine • Previous Articles     Next Articles

Induction of 3C protein of Coxsackievirus A16 in apoptosis of human rhabdomyosarcoma cells and its mechanism

Yingying SHI1,Shitong CHEN2,Bo HU2,Duo SHEN2,Kuang ZHU2,Siwei YE2,Jinmei FENG3()   

  1. 1.Department of Immunology,School of Medical Sciences,Jianghan University,Wuhan 430056,China
    2.School of Medical Sciences,Jianghan University,Wuhan 430056,China
    3.Department of Pathogen Biology,School of Medicsl Sciences,Jianghan University,Wuhan 430056,China
  • Received:2020-12-12 Online:2021-07-28 Published:2021-07-22
  • Contact: Jinmei FENG E-mail:fengjm@jhun.edu.cn

Abstract: Objective

To investigate the effect of Coxsackievirus A16 3C protein on the apoptosis of rhabdomyosarcoma RD cells, and to illuminate its possible mechanism.

Methods

The RD cells were cultured in vitro and divided into control group (transfected with 0.4 μg pCMV-HA), 0.1 μg pCMV-HA-3C group (transfected with 0.1 μg pCMV-HA-3C and 0.3 μg pCMV-HA), 0.2 μg pCMV-HA-3C group (transfected with 0.2 μg pCMV-HA-3C and 0.2 μg pCMV-HA),0.4 μg pCMV-HA-3C group (transfected with 0.4 μg pCMV-HA-3C) and protein kinase B(AKT) activator SC79 treatment group (pretreated with 4 mg·L-1 SC79 for 4 h, then transfected with 0.2 μg pCMV-HA-3C and 0.2 μg pCMV-HA).The survival rates of cells in various groups were detected by MTT assay,the apoptotic rates were detected by flow cytometry, the expression levels of Bcl-2 homologous antagonist /killer (Bak),Bcl-2 associated agonist of cell death (Bad) and p53 up-regulated modulator of apoptosis (PUMA) mRNA were detected by Real-time fluorescence quantitative PCR (RT-qPCR),and the expression levels of Bad, Bak, PUMA, PARP, cleaved-PARP, cleaved-caspase-3 and phosphorylated AKT (p-AKT) proteins in the RD cells in various groups were detected by Western blotting method.

Results

Compared with control group, the survival rate of the cells 0.4 μg pCMV-HA-3C group was significantly decreased (P<0.01). Compared with control group, the apoptotic rate of the cells in pCMV-HA-3C group was significantly increased (P<0.01). Compared with control group,the mRNA and protein expression levels of Bak, Bad and PUMA in 0.2 μg PCMV-HA-3C group were significantly increased (P<0.05), and the expression levels of cleaved-PARP and cleaved-caspase-3 proteins in different doses of pCMV-HA-3C groups were significantly increased (P<0.05).Compared with control group, the expression levels of cleaved-PARP and cleaved-caspase-3 proteins in 0.2 μg pCMV-HA-3C group were significantly increased (P<0.05),and the expression level of p-AKT protein was significantly decreased (P<0.01). Compared with 0.2 μg pCMV-HA-3C group,the the expression levels of cleaved-PARP,cleaved-caspase-3 and AKT proteins in the cells in SC79 treatment group were significantly decreased(P<0.05),and the p-AKT protein expression level was significantly increased (P<0.01).

Conclusion

Coxsackievirus A16 3C protein may induce the apoptosis of RD cells by inhibiting AKT signaling pathway.

Key words: Coxsackievirus A16, 3C protein, protein kinase B signaling pathway, apoptosis

CLC Number: 

  • R373.23