[Effect of fas mRNA expression on cytochrome C-induced apoptosis of HL-60 cells in vitro].
The aim of this study was to investigate the effect of cytochrome C on apoptosis of HL-60 cells and to explore the mechanism of HL-60 cell apoptosis induced by cytochrome C. The HL-60 cells were treated with different concentrations of cytochrome C for 24 hours. The concentrations of cytochrome C were as follows: 0 (control group), 9.375 mg/L, 18.75 mg/L, 37.5 mg/L, 75 mg/L and 150 mg/L. The apoptosis was analyzed by flow cytometry (FCM) after HL-60 cells were treated with different concentrations of cytochrome C for 24 hours. The fas mRNA expression changes of relevant apoptotic genes were examined by reverse transcription-polymerase chain reaction (RT-PCR) after HL-60 cells were treated with different concentrations of cytochrome C for 24 hours and were analyzed half-quantitatively. The expressions of fas involved in HL-60 treated with different concentrations of cytochrome C for 24 hours were detected by Western blot. The results indicated that the flow cytometry (FCM) (PI straining) showed obvious hypo-diploid peak before G(1) phase in the treated group, and its apoptotic ratio increased in a dose-dependent manner. The fas mRNA expression levels of the relevant apoptotic genes could be detected by RT-PCR after HL-60 cells treated with different concentrations of cytochrome C for 24 hours, and the expression of fas mRNA in HL-60 cells was increased by cytochrome C in dose-dependent manner within range of 0-37.5 mg/L. It is concluded that the cytochrome C up-regulates expressions of fas mRNA and their protein so as to induce apoptosis of the HL-60 cells.
Chen
Hua-Qin
HQ
Department of Pediatrics, Jiangmen Central Hosptal, Sun Yat-Sen University, Jiangmen 529030, China.
Chen
Ming-Zhen
MZ
Chen
Ri-Ling
RL
Fang
Xi-Min
XM
Xu
Xiao-Hong
XH
Feng
Hua-Jun
HJ
D017209:Apoptosis / Q000187:drug effects*; D045304:Cytochromes c / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D018922:HL-60 Cells; D006801:Humans; D012333:RNA, Messenger / Q000378:metabolism; D015854:Up-Regulation; D019014:fas Receptor / Q000378:metabolism*
C491901:FAS protein, human; D012333:RNA, Messenger; D019014:fas Receptor; D045304:Cytochromes c