Effects of chebulinic acid on differentiation of human leukemia K562 cells.
OBJECTIVE
To study effects of chebulinic acid on erythroid and megakaryocytic differentiation in K562 cells.


METHODS
The benzidine staining method was used to evaluate hemoglobin synthesis; the expression of erythroid specific glycophorin A (GPA) protein and megakaryocytic surface marker CD61 was determined by flow cytometry using fluorescence labeled antibodies; erythroid and megakaryocytic mRNA expression was analyzed by RT-PCR.


RESULTS
During erythroid differentiation induced by butyric acid (BA) or hemin, chebulinic acid not only inhibited the hemoglobin synthesis of BA- and hemin-treated K562 cells in concentration-dependent manner with IC50 of 4 micromol/L and 40 micromol/L respectively, but also inhibited another erythroid differentiation marker acetylcholinesterase at the concentration of 50 micromol/L in the cells either treated or untreated with each erythroid differentiation inducers, whereas chebulinic acid 50 micromol/L did not change GPA protein expression in these cells significantly. When K562 cells were treated with TPA 50 microg/L for 72 h to induce megakaryocytic differentiation, the presence of chebulinic acid 50 micromol/L slightly provoked the decrease of GPA protein expression induced by TPA. Chebulinic acid did not change the TPA-induced CD61 expression at the same concentration. Chebulinic acid also reduced the mRNA levels of erythroid relative genes including gamma-globin, PBGD, NF-E2, and GATA-1 genes in K562 cells either treated or untreated with BA, whereas chebulinic acid upregulated the mRNA levels of GATA-2 transcription factor in these cells.


CONCLUSIONS
Chebulinic acid had inhibitory effect on erythroid differentiation likely through changing transcriptional activation of differentiation relative genes, which suggests that chebulinic acid or other tannins might influence the efficiency of some anti-tumor drugs-induced differentiation or the hematopoiesis processes.
Yi
Zong-chun
ZC
Department of Biological Sciences and Biotechnology, Tsinghua University, Beijing 100084, China.
Wang
Zhao
Z
Li
Hai-xia
HX
Liu
Ming-jie
MJ
Wu
Rong-cong
RC
Wang
Xiao-hui
XH
D000110:Acetylcholinesterase / Q000096:biosynthesis / Q000235:genetics; D002454:Cell Differentiation / Q000187:drug effects; D004268:DNA-Binding Proteins / Q000096:biosynthesis / Q000235:genetics; D004912:Erythrocytes / Q000378:metabolism / Q000473:pathology; D050981:Erythroid-Specific DNA-Binding Factors; D050982:GATA1 Transcription Factor; D050989:GATA2 Transcription Factor; D005786:Gene Expression Regulation; D005914:Globins / Q000096:biosynthesis / Q000235:genetics; D006021:Glycophorins / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D047348:Hydrolyzable Tannins; D039661:Integrin beta3 / Q000096:biosynthesis* / Q000235:genetics; D020014:K562 Cells; D008533:Megakaryocytes / Q000378:metabolism / Q000473:pathology; D050983:NF-E2 Transcription Factor; D050984:NF-E2 Transcription Factor, p45 Subunit; D012333:RNA, Messenger / Q000235:genetics; D013634:Tannins / Q000494:pharmacology*; D014157:Transcription Factors / Q000096:biosynthesis / Q000235:genetics
D004268:DNA-Binding Proteins; D050981:Erythroid-Specific DNA-Binding Factors; D050982:GATA1 Transcription Factor; C494688:GATA1 protein, human; D050989:GATA2 Transcription Factor; C494711:GATA2 protein, human; D006021:Glycophorins; D047348:Hydrolyzable Tannins; C061248:ITGB3 protein, human; D039661:Integrin beta3; D050983:NF-E2 Transcription Factor; D050984:NF-E2 Transcription Factor, p45 Subunit; C494692:NFE2 protein, human; D012333:RNA, Messenger; D013634:Tannins; D014157:Transcription Factors; C103481:chebulinic acid; D005914:Globins; D000110:Acetylcholinesterase