[Rac1 expression and its effects on the cell cycle progression and apoptosis in human acute leukemic cell line HL-60].
The study was aimed to investigate the expression of Rac1 in human acute leukemic cell line HL-60 and effect of Rac1 on cell cycle progression and apoptosis. The mRNA expression of Rac1 in HL-60 cell line and normal human peripheral blood mononuclear cells (PBMNC) were examined by semi-quantitative RT-PCR. After transfection of HL-60 cells with different concentrations of Rac1 antisense oligodeoxynucleotide (ASODN) by means of FuGENE6, the survival, cell cycle, apoptosis of HL-60 cells were observed through MTT assay, FCM test, Wright-Giemsa, acridine orange/ethidium bromide (AO/EB) and Annexin V-FITC/PI staining test respectively. The results showed that Rac1 relative amount in HL-60 was 0.84 +/- 0.13, while it in the normal PBMNC was 0.26 +/- 0.1 (P < 0.01); the expression of Rac1 in HL-60 cells was significantly upregulated. Compared with sense oligodeoxynucleotide (SODN), HL-60 cell viability, after exposure to ASODN at a concentration of 2.0 g/L decreased, (73.7 +/- 5.0)% vs (93.2 +/- 3.0)% (P < 0.01), while the proportion of G(1) cells increased as (52.1 +/- 6.8)% vs (31.6 +/- 4.7)% (P < 0.05), the percentage of Annexin V positive cells increased, (19.2 +/- 2.1)% vs (4.1 +/- 1.7)% (P < 0.01), and HL-60 cells were observed to have formation of apoptotic bodies. The data presented indicate that Rac1 may be involved in regulation of HL-60 cell cycle and apoptosis, promote overproliferation of HL-60 cells and inhibit their apoptosis.
Gao
Guang-Xun
GX
Department of Hematology, Xijing Hospital, The Fourth Military Medical University, Xi'an 710033, China.
Chen
Xie-Qun
XQ
Zhang
Yong-Qing
YQ
Bai
Qing-Xian
QX
Huang
Gao-Sheng
GS
Zhang
Wei-Ping
WP
Liang
Rong
R
Dong
Bao-Xia
BX
Han
Dong-Mei
DM
Ge
Fan-Mei
FM
Wang
Zhe
Z
Zhu
Hua-Feng
HF
D017209:Apoptosis / Q000502:physiology*; D002453:Cell Cycle / Q000502:physiology*; D018922:HL-60 Cells; D006801:Humans; D016376:Oligonucleotides, Antisense / Q000096:biosynthesis* / Q000235:genetics; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D020830:rac1 GTP-Binding Protein / Q000096:biosynthesis* / Q000235:genetics / Q000502:physiology
D016376:Oligonucleotides, Antisense; D012333:RNA, Messenger; D020830:rac1 GTP-Binding Protein