Xu Jia-Wei, Jin Run-Ming, Wang Yan-Rong, Lin Wen, Meng Bing
Department of Pediatrics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China.
Zhongguo Dang Dai Er Ke Za Zhi. 2009 Apr;11(4):259-62.
Cardiotonic steroids (CTS) can bind to Na+, K+ -ATPase to activate complex intracellular signaling cascades regulating the proliferation and apoptosis of cells. The aim of this study was to investigate the effects of ouabain at different concentrations on growth regulation in various kinds of leukemia cell lines and explore the pathogenesis of leukemia, the functions of Na+, K+ -ATPase as a signal transduction conductor and its effects on cell growth.
Using the MTT assay, the survival rates of leukemia cell lines were observed 6, 12 and 24 hrs after treatment with 1 or 10 nmol/L ouabain. The expression of Na+, K+ -ATPase alpha1 subunit of leukemia cells was detected by Western blot.
The MTT results showed that ouabain at 1 nmol/L or 10 nmol/L induced proliferation of lymphocytic leukemia B95 and Jhhan cell lines, as well as megakaryocytic leukemia M07e and Meg01 cell lines. Ouabain at 1 nmol/L or 10 nmol/L increased the expression of Na+, K+ -ATPase alpha1 subunit. There were significant differences in the proliferation and the expression of Na+, K+ -ATPase alpha1 subunit of the leukemia cell lines between the ouabain treatment and the blank control groups 24 hrs after ouabain treatment (P<0.05). The proliferation effect of leukemia cell lines was in a direct proportion with the ouabain concentration and incubation time.
Na+, K+ -ATPase plays an important role in signal transductions. Through binding to ouabain, Na+, K+ -ATPase may regulate proliferation of leukemia cell lines of different origins. Ouabain at 1 nmol/L or 10 nmol/L may induce proliferation of lymphocytic leukemia cell lines (B95, Jhhan) and megakaryocytic leukemia cell lines (M07e, Meg01), and the proliferation effect was in a direct proportion with the concentration and incubation time of ouabain.
强心甾类化合物(CTS)可与Na⁺,K⁺ -ATP酶结合,激活复杂的细胞内信号级联反应,调节细胞的增殖和凋亡。本研究旨在探讨不同浓度哇巴因对各类白血病细胞系生长调控的影响,探索白血病的发病机制、Na⁺,K⁺ -ATP酶作为信号转导导体的功能及其对细胞生长的影响。
采用MTT法,观察1或10 nmol/L哇巴因处理白血病细胞系6、12和24小时后的存活率。通过蛋白质免疫印迹法检测白血病细胞中Na⁺,K⁺ -ATP酶α1亚基的表达。
MTT结果显示,1 nmol/L或10 nmol/L的哇巴因可诱导淋巴细胞白血病B95和Jhhan细胞系以及巨核细胞白血病M07e和Meg01细胞系的增殖。1 nmol/L或10 nmol/L的哇巴因可增加Na⁺,K⁺ -ATP酶α1亚基的表达。哇巴因处理24小时后,白血病细胞系的增殖及Na⁺,K⁺ -ATP酶α1亚基的表达在哇巴因处理组与空白对照组之间存在显著差异(P<0.05)。白血病细胞系的增殖效应与哇巴因浓度和孵育时间成正比。
Na⁺,K⁺ -ATP酶在信号转导中起重要作用。通过与哇巴因结合,Na⁺,K⁺ -ATP酶可能调节不同来源白血病细胞系的增殖。1 nmol/L或10 nmol/L的哇巴因可能诱导淋巴细胞白血病细胞系(B95、Jhhan)和巨核细胞白血病细胞系(M07e、Meg01)的增殖,且增殖效应与哇巴因的浓度和孵育时间成正比。