硒化壳聚糖对慢性粒细胞白血病K562细胞 bcr/abl融合基因表达的影响

目的:研究硒化壳聚糖对慢性粒细胞白血病K562细胞增殖的影响,探讨这种作用与bcr/abl融合基因表达的关系。方法:应用磺酰罗丹明B(SRB)法和集落形成法检测硒化壳聚糖对细胞增殖的影响;应用Western blot(免疫印迹)法检测细胞P210bcr/abl融合蛋白含量;应用反转录聚合酶链式反应(RT-PCR)法检测bcr/abl融合基因信使RNA(mRNA)水平。结果:50,100,200 mg·L-1硒化壳聚糖作用K562细胞48 h可明显抑制细胞增殖和降低其存活率(P<0.05,P<0.01);50,100,200 mg·L-1硒化壳聚糖作用48 h或200 mg·L-1硒化壳聚糖作用12,24,48 h后K562细胞内P210bcr/abl蛋白含量呈时间和剂量依赖性下降(P<0.05,P<0.01),而bcr/abl融合基因mRNA水平未见明显改变。结论:硒化壳聚糖可通过下调bcr/abl融合基因表达的P210bcr/abl融合蛋白对K562细胞增殖产生抑制作用。

Effect of Selenium Chitosan on Expression of bcr/abl Fusion Gene in Human Chronic Myelogenous Leukemia Cell Line K562

Objective: To investigate the effects of selenium chitosan (Sc) on proliferation of human chronic myelogenous leukemia cell line K562 and explore its relationship with the expression of bcr/abl fusion gene. Method: After treatment with Sc, cell proliferation was determined with SRB and clonal assay. The abundance of P210^bcr/abl protein was examined with Western blotting. The expression of bcr/abl fusion gene was detected by RT-PCR. Result: Exposure of K562 cells to 50, 100 and 200 mg ·L^-1 of Sc, respectively, for 48h could obviously inhibit proliferation and decrease their survival rate(P<0.05, P<0.01); After treatment with 50, 100, 200 mg ·L^-1 of Sc, respectively, for 48 h or 200 mg ·L^-1 of Sc for 12, 24, and 48 h, the abundance of P210^bcr/ablprotein in K562 cells was down-regulated in a concentration and time-dependent manner(P<0.05, P<0.01), but the expression of bcr/abl fusion gene was not changed. Conclusions: Sc could inhibit the proliferation of K562 cells cultured in vitro, and this effect was correlated with down-regulation of the abundance of P210^bcr/abl proteins encoded by bcr/abl fusion gene.