PMID- 24030155 OWN - NLM STAT- MEDLINE DCOM- 20140312 LR - 20211021 IS - 2041-4889 (Electronic) VI - 4 IP - 9 DP - 2013 Sep 12 TI - Overexpression of 4EBP1, p70S6K, Akt1 or Akt2 differentially promotes Coxsackievirus B3-induced apoptosis in HeLa cells. PG - e803-9 LID - 10.1038/cddis.2013.331 [doi] AB - Our previous studies have shown that the inhibition of phosphatidylinositol 3-kinase (PI3K) or mTOR complex 1 can obviously promote the Coxsackievirus B3 (CVB3)-induced apoptosis of HeLa cells by regulating the expression of proapoptotic factors. To further illustrate it, Homo sapiens eIF4E-binding protein 1 (4EBP1), p70S6 kinase (p70S6K), Akt1 and Akt2 were transfected to HeLa cells, respectively. And then, we established the stable transfected cell lines. Next, after CVB3 infection, apoptosis in different groups was determined by flow cytometry; the expressions of Bim, Bax, caspase-9 and caspase-3 were examined by real-time fluorescence quantitative PCR and western blot analysis; the expression of CVB3 mRNA and viral capsid protein VP1 were also analyzed by real-time fluorescence quantitative PCR, western blot analysis and immunofluorescence, respectively. At the meantime, CVB3 replication was observed by transmission electron microscope. We found that CVB3-induced cytopathic effect and apoptosis in transfected groups were more obvious than that in controls. Unexpectedly, apoptosis rate in Akt1 group was higher than others at the early stage after viral infection and decreased with the viral-infected time increasing, which was opposite to other groups. Compared with controls, the expression of CVB3 mRNA was increased at 3, 6, 12 and 24 h postinfection (p. i.) in all groups. At the meantime, VP1 expression in 4EBP1 group was higher than control during the process of infection, while the expressions in the other groups were change dynamically. Moreover, overexpression of 4EBP1 did not affect the mRNA expressions of Bim, Bax, caspase-9 and caspase-3; while protein expressions of Bim and Bax were decreased, the self-cleavages of caspase-9 and caspase-3 were stimulated. Meanwhile, overexpression of p70S6K blocked the CVB3-induced Bim, Bax and caspase-9 expressions but promoted the self-cleavage of caspase-9. In the Akt1 group, it is noteworthy that the expressions of Bim protein were higher than controls at 3 and 6 h p. i. but lower at 24 h p. i., and the expression of Bax protein were higher at 6 and 24 h p. i., while their mRNA expressions were all decreased. Furthermore, overexpression of Akt1 stimulated the procaspase-9 and procaspase-3 expression but blocked their self-cleavages. Overexpression of Akt2, however, had little effect on Bim, Bax and caspase-3, while prevented caspase-9 from self-cleavage at the late stage of CVB3 infection. As stated above, our results demonstrated that overexpression of 4EBP1, p70S6K, Akt1 or Akt2 could promote the CVB3-induced apoptosis in diverse degree via different mediating ways in viral replication and proapoptotic factors in BcL-2 and caspase families. As 4EBP1, p70S6K and Akt are the important substrates of PI3K and mammalian target of rapamycin (mTOR), we further illustrated the role of PI3K/Akt/mTOR signaling pathway in the process of CVB3-induced apoptosis. FAU - Li, X AU - Li X AD - Department of Pediatrics, The Third Xiangya Hospital, Central South University, Changsha, China. FAU - Li, Z AU - Li Z FAU - Zhou, W AU - Zhou W FAU - Xing, X AU - Xing X FAU - Huang, L AU - Huang L FAU - Tian, L AU - Tian L FAU - Chen, J AU - Chen J FAU - Chen, C AU - Chen C FAU - Ma, X AU - Ma X FAU - Yang, Z AU - Yang Z LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20130912 PL - England TA - Cell Death Dis JT - Cell death & disease JID - 101524092 RN - 0 (Adaptor Proteins, Signal Transducing) RN - 0 (Apoptosis Regulatory Proteins) RN - 0 (BCL2L11 protein, human) RN - 0 (Bcl-2-Like Protein 11) RN - 0 (Cell Cycle Proteins) RN - 0 (EIF4EBP1 protein, human) RN - 0 (Membrane Proteins) RN - 0 (Phosphoproteins) RN - 0 (Proto-Oncogene Proteins) RN - 0 (RNA, Messenger) RN - 0 (Viral Proteins) RN - 0 (bcl-2-Associated X Protein) RN - EC 2.7.11.1 (AKT1 protein, human) RN - EC 2.7.11.1 (AKT2 protein, human) RN - EC 2.7.11.1 (Proto-Oncogene Proteins c-akt) RN - EC 2.7.11.1 (Ribosomal Protein S6 Kinases, 70-kDa) RN - EC 3.4.22.- (Caspase 3) RN - EC 3.4.22.- (Caspase 9) SB - IM MH - Adaptor Proteins, Signal Transducing/*metabolism MH - *Apoptosis MH - Apoptosis Regulatory Proteins/metabolism MH - Bcl-2-Like Protein 11 MH - Caspase 3/genetics/metabolism MH - Caspase 9/genetics/metabolism MH - Cell Cycle Proteins MH - Cytopathogenic Effect, Viral MH - Enterovirus/genetics/*physiology/ultrastructure MH - Gene Expression Regulation, Viral MH - HeLa Cells MH - Humans MH - Membrane Proteins/metabolism MH - Models, Biological MH - Phosphoproteins/*metabolism MH - Proto-Oncogene Proteins/metabolism MH - Proto-Oncogene Proteins c-akt/*metabolism MH - RNA, Messenger/genetics/metabolism MH - Ribosomal Protein S6 Kinases, 70-kDa/*metabolism MH - Transfection MH - Viral Proteins/metabolism MH - bcl-2-Associated X Protein/metabolism PMC - PMC3789189 EDAT- 2013/09/14 06:00 MHDA- 2014/03/13 06:00 PMCR- 2013/09/01 CRDT- 2013/09/14 06:00 PHST- 2013/05/20 00:00 [received] PHST- 2013/08/04 00:00 [revised] PHST- 2013/08/06 00:00 [accepted] PHST- 2013/09/14 06:00 [entrez] PHST- 2013/09/14 06:00 [pubmed] PHST- 2014/03/13 06:00 [medline] PHST- 2013/09/01 00:00 [pmc-release] AID - cddis2013331 [pii] AID - 10.1038/cddis.2013.331 [doi] PST - epublish SO - Cell Death Dis. 2013 Sep 12;4(9):e803-9. doi: 10.1038/cddis.2013.331.