ANTISENSE OLIGONUCLEOTIDE INHIBITION OF ACETYLCHOLINESTERASE GENE-EXPRESSION INDUCES PROGENITOR-CELL EXPANSION AND SUPPRESSES HEMATOPOIETIC APOPTOSIS EX-VIVO

被引:97
作者
SOREQ, H
PATINKIN, D
LEVLEHMAN, E
GRIFMAN, M
GINZBERG, D
ECKSTEIN, F
ZAKUT, H
机构
[1] MAX PLANCK INST EXPTL MED,D-37075 GOTTINGEN,GERMANY
[2] TEL AVIV UNIV,SACKLER FAC MED,EDITH WOLFSON MED CTR,DEPT OBSTET & GYNECOL,IL-58100 HOLON,ISRAEL
关键词
D O I
10.1073/pnas.91.17.7907
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
To examine the role of acetylcholinesterase (EC 3.1.1.7) in hematopoietic cell proliferation and differentiation, we administered a 15-mer phosphorothioate oligonucleotide, antisense to the corresponding ACHE gene (AS ACHE), to primary mouse bone marrow cultures. Within 2 hr of AS-ACHE addition to the culture, ACHE mRNA revels dropped by approximate to 90%, as compared with those in cells treated with the ''sense'' oligomer, S-ACHE. Four days after AS-ACHE treatment, ACHE mRNA increased to levels 10-fold higher than in S-ACHE cultures or in fresh bone marrow. At this later time point, differential PCR display revealed significant differences between cellular mRNA transcripts in bone marrow and those in AS-ACHE- or S-ACHE-treated cultures. These oligonucleotide triggered effects underlay considerable alterations at the cellular level: AS-ACHE but not S-ACHE increased cell counts, reflecting enhanced proliferation, In the presence of erythropoietin it also enhanced colony counts, reflecting expansion of progenitors. AS-ACHE further suppressed apoptosis-related fragmentation of cellular DNA in the progeny cells, and it diverted hematopoiesis toward production of primitive blasts and macrophages in a dose-dependent manner promoted by erythropoietin. These findings suggest that the hematopoietic role of acetylcholinesterase, anticipated to be inverse to the observed antisense effects, is to reduce proliferation of the multipotent stem cells committed to erythropoiesis and megakaryocytopoiesis and macrophage production and to promote apoptosis in their progeny. Moreover, these findings may explain the tumorigenic association of perturbations in ACHE gene expression with leukemia.
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页码:7907 / 7911
页数:5
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