[Preliminary study on leukemia related gene zo-1 involved in pathogenesis of leukemia].
The study was aimed to identify a new leukemia related gene zo-1 from leukemia and to explore its mechanism in leukemia. Methylation specific PCR (MSP) was used for testing gene zo-1 methylation in leukemia cells. The gene zo-1 specific siRNA was designed according to its sequence, and transfected into THP-1 cell, and the cells were cultured for 48 hours before harvesting. The effect of zo-1 siRNA was monitored by RT-PCR. The cellular proliferation activity was assayed by CCK-8, the apoptosis was detected by Annexin-V-fluorescence in isothiocyanate (FITC) assay, and cell cycle was observed by propidium iodide (PI). The results indicated that the gene zo-1 in patients with acute leukemia was hypermethylated, while the gene zo-1 in healthy persons was unmethylated. The THP-1 cells with unmethylation of zo-1 gene promoter overexpressed the gene zo-1, while the Molt4 and HL-60 cells with hypermethylation of gene zo-1 promoter did not express the gene zo-1. The silenced zo-1 gene in Molt4 and HL-60 leukemia cell lines could be reactivated by demethylation treatment with 5-AZA-dC. The oligofectamine-transfected siRNA for zo-1 gene successfully inhibited the expression of gene zo-1 in THP-1 cells, but did not interfere with cell proliferation, cell cycle and apoptosis. It is concluded that gene zo-1 is a leukemia-related gene. Gene zo-1 in acute leukemia was hypermethylated, the methylation status of gene zo-1 regulates the expression of gene zo-1. Lack of gene zo-1 expression in THP-1 cells does not influence the cell proliferation, apoptosis and cell cycle, which suggests that the methylation of gene zo-1 may be involved in the genesis of acute leukemia, its mechanism is worthy to be studied.
Dou
Li-Ping
LP
Department of Hematology, Chinese PLA General Hospital, Beijing 100853, China.
Liu
Jun-Hua
JH
Wang
Chang
C
Zhao
Yu
Y
Wang
Quan-Shun
QS
Liu
Jing-Hua
JH
Liu
Chun-Hui
CH
Lou
Fang-Ding
FD
Yu
Li
L
D018899:CpG Islands; D019175:DNA Methylation; D020868:Gene Silencing; D018922:HL-60 Cells; D006801:Humans; D007938:Leukemia / Q000235:genetics*; D008565:Membrane Proteins / Q000378:metabolism*; D010750:Phosphoproteins / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000235:genetics; D062826:Zonula Occludens-1 Protein
D008565:Membrane Proteins; D010750:Phosphoproteins; D034741:RNA, Small Interfering; C569042:TJP1 protein, human; D062826:Zonula Occludens-1 Protein