[Two-dimensional polyacrylamide gel electrophoresis analysis of apoptosis in K562 cells induced by harringtonine].
OBJECTIVE
Harringtonine (HT),an anti-tumor drug,has been widely used to treat acute or chronic myeloid leukemia,and obtained satisfactory effects. Studies showed that the anti-tumor activity of HT is related with the apoptosis-inducing effect,but the molecular mechanisms remain unclear. This study was to analyze the protein maps contributed to the apoptosis in K562 cells induced by HT,and to screen the apoptosis-associated proteins.


METHODS
Flow cytometry was used to distinguish K562 cells of early apoptosis stage from those of late apoptosis stage through Annexin V and PI staining. Two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) was used to separate and compare the HT-induced apoptotic K562 cells and control K562 cells.


RESULTS
When K562 cells were treated with 10 microg/ml HT for 5,a nd 24 hours,percentages of early apoptotic cells (Annexin V+/PI-) were 28.3%, and 18.1%(P< 0.01), while percentages of late apoptotic cells (Annexin V+/PI+) were 9.1%,and 20.2% (P< 0.01), respectively. Statistical analysis showed (1300+/-50) protein spots were resolved with a match rate of (88.3+/-2.0)% in control K562 cells. Ten protein spots in late apoptotic cells displayed changes in expression after induction of HT for 24 hours (P< 0.01), of which 8 showed higher expression, 1 decreased,and 1 only expressed in control k562 cells.


CONCLUSIONS
These proteins may be apoptosis-associated proteins of K562 cells induced by HT.
Department of Hematology, Nanfang Hospital, The First Military Medical University, Guangzhou, Guangdong, 510515, P.R china.
Rong
R
Li
Xiao-Li
XL
Liu
Qing-Feng
QF
Du
Song
S
Zhang
Shu-Yun
SY
Zhou
D000972:Antineoplastic Agents, Phytogenic / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D015180:Electrophoresis, Gel, Two-Dimensional; D006248:Harringtonines / Q000494:pharmacology*; D006801:Humans; D020014:K562 Cells; D025941:Protein Interaction Mapping; D020543:Proteome / Q000737:chemistry*
D000972:Antineoplastic Agents, Phytogenic; D006248:Harringtonines; D020543:Proteome