摘要
目的观察在低剂量蟾蜍灵联合全反式维甲酸（ATRA）诱导HL-60细胞向粒-单核细胞分化过程中细胞黏附功能的变化，检测黏附相关蛋白Pyk2及其底物蛋白Paxillin表达的变化，并探讨蛋白酶体通路对细胞黏附功能以及Pyk2和Paxillin表达的影响。方法采用流式细胞术检测细胞CD11b的表达；采用MTF法检测细胞黏附能力；采用Westernblot技术检测Pyk2、Paxillin和微管蛋白（Tubulin）的表达水平。结果低剂量蟾蜍灵＋ATRA联合用药以时间依赖性的方式诱导HL-60细胞分化，CD11b阳性率在处理2d和4d分别为（20．0±2．8）％和（75．0±5．3）％，并伴有细胞黏附功能的上调。同时Pyk2和Paxillin表达水平的上调也呈时间依赖性。蛋白酶体抑制剂硼替佐米以剂量依赖性的方式抑制细胞黏附能力，0、1和10nmol／L硼替佐米处理的HL-60细胞的黏附水平分别为（9．4±0．8）％、（7．8±0．1）％和（5．3±0．3）％，硼替佐米处理组同时伴有Pyk2表达水平的下调，但Paxillin的表达不受影响。结论Pyk2参与细胞黏附功能的调节；蛋白酶体通路抑制剂PS-341可能通过下调Pyk2表达，抑制HL-60细胞黏附功能。
Objective To investigate changes in the adherent ability, the expression of adhesion related proteins Pyk2 and paxillin during HL-60 cells differentiation into granulocyte-monocyte induced by lowdose （LD） bufalin in combination with all-trans retinoic acid （ ATRA）, and to explore the effects of bortezomib on cellular adhesion and the expression of Pyk2 and paxillin. Methods The expression of CDllb was detected by flow cytometry, cellular adherence ability by MTT assay, and the expressions of Pyk2, paxillin and tubulin by Western blot. Results The combination of 5 nmol/L bufalin and 30 nmol/L ATRA induced HL-60 cells differentiation in a time-dependent manner,the percentages of CDllb positive cells treated for 2 d and 4 d being （20.0 ± 2.8） % and （75.0 ± 5.3 ） %, respectively, with the increasing of cellular adherence ability. Meanwhile the expressions of Pyk2 and Paxillin were also up-regulated in a time-dependent manner. Bort- ezomib suppressed HL-60 cell adhesion in a dose-dependent manner. At concentrations of 1 nmol/L and 10 nmol/L the adherence level were （7.8 ± 0.1 ） % and （5.3 ± 0.3 ） %, respectively, with down-regulation of Pyk2 but not Paxillin. Conclusion Pyk2 is involved in the regulation of cellular adherence function. Bortezomib might inhibit HL-60 cells adhension function by down-regulation of Pyk2 expression.
出处
《中华血液学杂志》
CAS
CSCD
北大核心
2008年第12期828-831,共4页
Chinese Journal of Hematology
基金
基金项目：国家自然科学基金（30770993、30700807）
辽宁省科学技术厅基金（2004225004-11）
