PMID- 27667455
OWN - NLM
STAT- MEDLINE
DCOM- 20170629
LR  - 20170629
IS  - 1007-8738 (Print)
IS  - 1007-8738 (Linking)
VI  - 32
IP  - 10
DP  - 2016 Oct
TI  - [Protein kinase A inhibitor H-89 blocks polyploidization of SP600125-induced CMK 
      cells by regulating phosphorylation of ribosomal protein S6 kinase 1].
PG  - 1321-1326
AB  - Objective To investigate the regulatory effect of post-translation modification 
      of ribosomal protein S6 kinase 1 (S6K1) on the polyploidization of 
      megakaryocytes. Methods SP600125, a c-Jun N-terminal kinase (JNK) inhibitor, and 
      H-89, a cAMP-dependent protein kinase (PKA) inhibitor, were used to treat CMK 
      cells separately or in combination. With propidium iodide (PI) to dye DNA in the 
      treated cells, the relative DNA content was detected by flow cytometry, and then 
      the DNA polyploidy was analyzed. The change of expression and phosphorylation of 
      ribosomal protein S6 kinase 1 (S6K1), an important mammalian target of rapamycin 
      (mTOR) downstream target molecule, was analyzed by Western blotting. Molecular 
      docking study and kinase activity assay were performed to analyze the combination 
      of H-89 with S6K1 and the effect of H-89 on the activity of S6K1 kinase. Results 
      SP600125 induced CMK cell polyploidization in a time-dependent and dose-dependent 
      manner. At the same time, it increased the phosphorylation of S6K1 at 
      Thr421/Ser424 and decreased the phosphorylation of S6K1 at Thr389. H-89 not only 
      blocked polyploidization, but also decreased the phosphorylation of S6K1 at 
      Thr421/Ser424 and increased the phosphorylation of S6K1 at Thr389. Molecular 
      docking and kinase activity assay showed that H-89 occupied the ATP binding sites 
      of S6K1 and inhibited its activity. Noticeably, both H-89 and SP600125 inhibited 
      the activity of PKA. Moreover, the two drugs further inhibited the activity of 
      PKA when used together. Therefore, these data indicated that H-89 blocked the 
      SP600125-induced polyploidization of CMK cells mainly by changing S6K1 
      phosphorylation state, rather than its inhibitory effect on PKA. Conclusion H-89 
      can block the polyploidization of SP600125-induced CMK cells by regulating S6K1 
      phosphorylation state.
FAU - Zhao, Song
AU  - Zhao S
AD  - Department of Experimental Medicine, General Hospital, Shenyang Military Area 
      Command, Shenyang 110016, China.
FAU - Yang, Jingang
AU  - Yang J
AD  - Department of Experimental Medicine, General Hospital, Shenyang Military Area 
      Command, Shenyang 110016, China.
FAU - Li, Changling
AU  - Li C
AD  - Department of Experimental Medicine, General Hospital, Shenyang Military Area 
      Command, Shenyang 110016, China.
FAU - Xing, Sining
AU  - Xing S
AD  - Department of Experimental Medicine, General Hospital, Shenyang Military Area 
      Command, Shenyang 110016, China.
FAU - Yu, Ying
AU  - Yu Y
AD  - Department of Experimental Medicine, General Hospital, Shenyang Military Area 
      Command, Shenyang 110016, China.
FAU - Liu, Shuo
AU  - Liu S
AD  - Department of Experimental Medicine, General Hospital, Shenyang Military Area 
      Command, Shenyang 110016, China.
FAU - Pu, Feifei
AU  - Pu F
AD  - Department of Experimental Medicine, General Hospital, Shenyang Military Area 
      Command, Shenyang 110016, China.
FAU - Ma, Dongchu
AU  - Ma D
AD  - Department of Experimental Medicine, General Hospital, Shenyang Military Area 
      Command, Shenyang 110016, China. *Corresponding author, E-mail: 
      mdc580819@sina.com.
LA  - chi
PT  - Journal Article
PL  - China
TA  - Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi
JT  - Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular 
      immunology
JID - 101139110
RN  - 0 (Anthracenes)
RN  - 0 (Isoquinolines)
RN  - 0 (Protein Kinase Inhibitors)
RN  - 0 (Sulfonamides)
RN  - 1TW30Y2766 (pyrazolanthrone)
RN  - EC 2.7.11.1 (Ribosomal Protein S6 Kinases, 90-kDa)
RN  - EC 2.7.11.1 (ribosomal protein S6 kinase 1, human)
RN  - M876330O56 (N-(2-(4-bromocinnamylamino)ethyl)-5-isoquinolinesulfonamide)
SB  - IM
MH  - Anthracenes/chemistry/*pharmacology
MH  - Cell Line
MH  - Humans
MH  - Isoquinolines/chemistry/*pharmacology
MH  - Megakaryocytes/*cytology/drug effects/*enzymology/metabolism
MH  - Molecular Docking Simulation
MH  - Phosphorylation
MH  - Polyploidy
MH  - Protein Kinase Inhibitors/chemistry/*pharmacology
MH  - Ribosomal Protein S6 Kinases, 90-kDa/chemistry/genetics/*metabolism
MH  - Sulfonamides/chemistry/*pharmacology
EDAT- 2016/09/27 06:00
MHDA- 2017/07/01 06:00
CRDT- 2016/09/27 06:00
PHST- 2016/09/27 06:00 [entrez]
PHST- 2016/09/27 06:00 [pubmed]
PHST- 2017/07/01 06:00 [medline]
PST - ppublish
SO  - Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2016 Oct;32(10):1321-1326.