丙戊酸对HL-60/HT细胞P27^(Kip1)、P170表达水平及耐药性的影响  

Effect of valproic acid on the expression of P27^(Kip1) and P170 and drug resistance of HL-60/HT cells

作　　者：李益清[1] 尹松梅[1] 谢双锋[1] 马丽萍[1] 聂大年[1] 王秀菊[1] 吴裕丹[1] 

机构地区：[1]中山大学附属第二医院血液内科,广东广州510120

出　　处：《南方医科大学学报》2009年第3期423-427,共5页Journal of Southern Medical University

基　　金：广东省自然科学基金(06021324);广州市科技计划(2006J1-C0071)

摘　　要：目的观察丙戊酸对白血病细胞P27Kip1、P170表达水平和耐药性的影响,探讨其可能的作用机制。方法递增压力选择培养法建立白血病耐药细胞株HL-60/HT,MTT法检测丙戊酸作用前后细胞的增殖情况及对Ara-C耐药性的改变,流式细胞术测定HL-60细胞、HL-60/HT细胞中P27Kip1、P170的表达和细胞周期。结果成功诱导培养建立耐药HL-60/HT细胞,其对HT、VCR、DNR、Ara-C的耐药倍数分别为9.30、5.20、4.91、3.65倍,耐药性能稳定。HL-60/HT细胞P27Kip1表达水平明显低于正常人单个核细胞及HL-60细胞(P<0.05),P170表达水平则明显高于正常人单个核细胞及HL-60细胞(P<0.05)。丙戊酸+Ara-C联合用药对HL-60、HL-60/HT细胞的生长抑制率明显高于单独用药的丙戊酸组和Ara-C组,q值分别为1.37和1.51,说明合用具有协同作用。丙戊酸作用于HL-60、HL-60/HT细胞后,细胞中P27Kip1表达水平、G1期细胞比加药前增加(P<0.05);P170表达水平在加药前后无明显变化(P>0.05)。结论耐药白血病HL-60/HT细胞中P27Kip1的表达低于敏感细胞HL-60,丙戊酸能抑制HL-60/HT细胞增殖,并降低其对Ara-C的耐药性。作用机制可能与改变P27Kip1的表达、增加G1期细胞含量有关,与P170作用无关。Objective To investigate the effect ofvalproic acid on the expression of P27^Kip1 and P170 and drug resistance of leukemia HL60/HT cell line and explore its possible mechanisms. Methods HL-60/FIT cells were derived from HL-60 cells induced by harringtonine （HT） in gradient concentrations. The inhibitory effect of valproic acid on the proliferation of HL-60 and HL-60/HT cells was evaluated by MTT assay, and the P27^Kip1 expression, P170 expression and cell cycle of the cells were analyzed with flow cytometry. Results The multidrug-resistant HL-60/HT was acquired, which showed a stable drug-resistant index with increased IC50 of HT, VCR, DNR and Ara-c by 9.30, 5.20, 4.91 and 3.65 folds, respectively, as compared with those of HL60 cells. The expression of P27^Kip1 in HL-60/HT cells was significantly lower but P170 expression significantly higher than that of HL-60 cells and normal mononuclear cells （P〈0.05）. The expressions of P27^Kip1 and P170 showed no significant difference between normal mononuclear cells and HL-60 cells. The growth inhibition rate of VPA combined with Ara-C was significantly higher than that ofvalproic acid or Ara：C alone in HL-60/HT cells and HL-60 cells （q=1.37 and 1.51, respectively）. HL-60/HT and HL-60 cells cultured in the presence of VPA resulted in a significant increase in the expression of P27^Kip1 and the G1-phase Cells （P〈0.05）, but the expression of P170 underwent no significant changes （P〉0.05）. Conclusion HL-60/HT cells have lower P27^Kip1 expression compared with HL-60 cells. Valproic acid can inhibit the growth of HL-60/HT cells and enhance their Ara-C sensitivity possibly by increasing P27^Kip1 expression and causing cell cycle arrest in G1 phase.

关 键 词：丙戊酸 HL-60/HT细胞 P27^KIP1 多药耐药 P170 

分 类 号：R733.7[医药卫生—肿瘤]