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Archive > Volume 24 Issue 8 > 2017,24(8):828-832. DOI:10.3872/j.issn.1007-385X.2017.08.002 Prev Next
Prompt Report
Constitutively active Rac1 GTPase confers leukemia cells quiescence through upregulating myeloproliferative leukemia
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Abstract:
Objective: To determine the role of Rac1 GTPase activation in the regulation of leukemia cell quies-cence and investigate the possible mechanism. Methods: Constitutively active Rac1 lenti-virus vector was construct-ed to transfect KG-1a leukemia cells (Rac1-V12-KG-1a). The G0 phase cell ratio in sorted Rac1-V12-KG-1a and KG-1a cells transfected with empty vector (pCDH-KG-1a) was compared. Furthermore, after 4 days treatment with NSC23766, the Rac1-specific inhibitor, the G0 phase cell ratio in KG-1a cells was also detected. The expression lev-els of cell quiescence and cell cycle associated moleculars were then determined by RT-PCR. The G0 phase cell ra-tio 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-1a cells ([15.30±0.60]% vs [11.50 ± 0.17]%, P<0.05). After the treatment of Rac1-specific inhibitor, the G0 phase cell ratio of the KG-1a cells was decreased significantly (P<0.05), and the effect was concentration depen-dent. RT-PCR showed that mRNA transcription levels of cell cycle related factors (P21 ， P27 ， P57) were up-regulat-ed in Rac1-V12-KG-1a cells at different level; moreover, the expressions of N-Cadherin and MPL were also signifi-cantly higher in Rac1-V12-KG-1a 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 Rac1 GTPase could increase the ratio of leukemia cells at G0 phase, and promote leukemia cells quiescence by up-regulating MPL.
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Project Supported:
Project supported by the National Natural Science Foundation of China （No. 81370599）
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