不同浓度哇巴因对白血病细胞增殖的影响

Effects of ouabain at different concentrations on growth of leukemia cells

摘要:

目的：近年来研究表明钠钾ATP酶通过与强心甾类固醇（CTS）特异性结合，可激活细胞内一系列信号级联反应，调节细胞的增殖与凋亡。该研究通过检测不同浓度的哇巴因对多种白血病细胞株增殖的影响，进一步探讨白血病的发病机制及钠钾ATP酶作为信号转导体的功能效应以及其在细胞增殖调控过程中的作用。方法：采用MTT方法检测不同浓度的哇巴因（1 nmol/L、10 nmol/L）在不同作用时间（6 h，12 h，24 h）下对多种白血病细胞株增殖的作用，Western blot检测白血病细胞株钠钾ATP酶α1亚单位表达水平的变化情况。结果：MTT结果表明1 nmol/L、10 nmol/L哇巴因可促进急性淋巴细胞白血病细胞株B95，Jhhan与巨核系白血病M07e，Meg01细胞株增殖，Western blot结果显示1 nmol/L、10 nmol/L哇巴因可使钠钾ATP酶α1亚单位表达升高。与空白对照组相比，1 nmol/L、10 nmol/L哇巴因作用24 h时细胞增殖程度与钠钾ATP酶α1亚单位表达升高，差异具有显著性（P<0.05）。细胞增殖作用与哇巴因浓度与孵育时间成正比。结论： 钠钾ATP酶具有信号传导功能，它能通过与哇巴因结合调节多种不同来源白血病细胞株的增殖。1 nmol/L与10 nmol/L的哇巴因可促进白血病细胞株B95、Jhhan、M07e、Meg01增殖，增殖效应与哇巴因浓度与孵育时间呈正比。［中国当代儿科杂志，2009，11（4）：259-262］

Abstract:

OBJECTIVE: 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. METHODS: 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 α1 subunit of leukemia cells was detected by Western blot. RESULTS: 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 α1 subunit. There were significant differences in the proliferation and the expression of Na+, K+-ATPase α1 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. CONCLUSIONS: 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.［Chin J Contemp Pediatr, 2009, 11 (4):259-262］