中国肿瘤生物治疗杂志2017，Vol.24Issue(8)：828-832,5.DOI:10.3872/j.issn.1007-385x.2017.08.002
组成活化型Rac1 GTP酶通过上调促血小板血成素受体的表达促进白血病细胞的静息
Constitutively active Rac1 GTPase confers leukemia cells quiescence through upregulating myeloproliferative leukemia
摘要
Abstract
Objective:To determine the role of Racl GTPase activation in the regulation of leukemia cell quiescence and investigate the possible mechanism.Methods:Constitutively active Rac 1 lenti-virus vector was constructed to transfect KG-1a leukemia cells (Rac1-V12-KG-1a).The G0 phase cell ratio in sorted Racl-V12-KG-la and KG-1 a cells transfected with empty vector (pCDH-KG-1 a) was compared.Furthermore,after 4 days treatment with NSC23766,the Racl-specific inhibitor,the G0 phase cell ratio in KG-1a cells was also detected.The expression levels of cell quiescence and cell cycle associated moleculars were then determined by RT-PCR.The G0 phase cell ratio of MPL and MPL+ leukemia cells in AML1-ETO9a leukemia mouse was detected by flow cytometry.Results:The results showed that the G0 phase cell ratio in Rac1-V12-KG-1a cells was significantly higher than that in pCDH-KG-1 a cells ([15.30±0.60]％ vs [11.50 ± 0.17]％,P＜0.05).After the treatment of Rac 1-specific inhibitor,the G0 phase cell ratio of the KG-1 a cells was decreased significantly (P＜0.05),and the effect was concentration dependent.RT-PCR showed that mRNA transcription levels of cell cycle related factors (P21,P27,P57) were up-regulated in Rac 1-V 12-KG-1 a cells at different level;moreover,the expressions of N-Cadherin and MPL were also significantly higher in Racl-V12-KG-la cells (P＜0.05).Flow cytometry analysis showed that the percentage of G0 phase MPL+ leukemia cells was significantly higher than that of MPL leukemia cells ([40.3±3.5])％ vs [19.05±7.65]％,P＜0.05).Conclusion:Activation of Racl GTPase could increase the ratio of leukemia cells at G0 phase,and promote leukemia cells quiescence by up-regulating MPL.关键词
Rac1 GTP酶/白血病细胞/静息/促血小板生成素受体/微环境Key words
Rac1 GTPase/leukemia cell/quiescence/myeloproliferative leukemia (MPL)/microenvironment分类
医药卫生引用本文复制引用
李欢,王继英,于沛,陈树英,邢海燕,田征,唐克晶,王敏,饶青..组成活化型Rac1 GTP酶通过上调促血小板血成素受体的表达促进白血病细胞的静息[J].中国肿瘤生物治疗杂志,2017,24(8):828-832,5.基金项目
国家自然科学基金资助项目(No.81370599).Project supported by the National Natural Science Foundation of China(No.81370599) （No.81370599）
