HOXA10基因沉默对白血病细胞化疗药物敏感性影响的观察  

Effect of silencing HOXA10gene on chemotherapeutic drug sensitivity of leukemic cells

作　　者：贾秀红[1] 张艳君 李建厂[1] 

机构地区：[1]滨州医学院附属医院儿科,山东滨州256603

出　　处：《中华肿瘤防治杂志》2013年第18期1405-1409,共5页Chinese Journal of Cancer Prevention and Treatment

基　　金：山东省科学技术发展计划(2010GSF10264);山东省自然科学基金(Y2007C018)

摘　　要：目的:探讨沉默同源盒基因A10(HOXA10)对白血病U937细胞化疗药物敏感性的影响。方法:选择白血病细胞株U937为实验细胞,应用慢病毒载体介导短发卡RNA(shRNA)靶向沉默HOXA10基因(KD组)、并设阴性对照(NC)组、空白对照(CON)组。用流式细胞仪测定慢病毒的感染效率,RT-PCR和蛋白质印迹法检测shRNA干扰后的HOXA10基因mRNA和蛋白表达水平;在阿糖胞苷和柔红霉素作用实验细胞后,用四甲基噻唑氮蓝(methyl thiazolyl tetrazolium,MTT)法和流式细胞仪分别检测实验细胞的增殖和凋亡水平。结果:慢病毒对U937细胞的感染率为90%;KD组HOXA10mRNA表达水平为0.047 3±0.016 8,低于NC组0.225 2±0.059 5,P=0.004,同时低于CON组0.261 3±0.054 9,P=0.002;KD组HOXA10蛋白表达水平为0.001 5±0.000 8,低于NC组0.013 4±0.005 9,P=0.001,同时低于CON组0.014 5±0.001 6,P=0.001。KD组U937细胞的阿糖胞苷半数抑制浓度(IC50)为(12.80±4.32)μg/mL,显著低于NC组(116.65±16.66)μg/mL,P<0.001,同时低于CON组(100.14±15.45)μg/mL,P<0.001;KD组U937细胞柔红霉素IC50为(75.33±20.40)ng/mL,低于NC组(245.00±46.13)ng/mL,P=0.001,同时低于CON组(267.67±32.65)ng/mL,P<0.001。KD组阿糖胞苷诱导的细胞凋亡率为(71.37±3.24)%,高于NC组(45.27±1.88)%,P<0.001,同时高于CON组(39.39±4.46)%,P<0.001;KD组柔红霉素诱导的细胞凋亡率为(59.72±3.07)%,高于NC组(34.96±5.89)%,P=0.004,同时高于CON组(35.05±9.23)%,P=0.004。NC和CON组之间IC50和细胞凋亡率差异无统计学意义。结论:沉默HOXA10基因可增加U937细胞对化疗药物的敏感性。OBJECTIVE： To investigate the effects of silencing HOXA10 gene on the chemotherapeutic drugs sensi- tivity of U937 cells. METHODS： U937 cells were divided into interference group （Lentivirus-mediated short-hairpin RNA targeting of HOXA10 gene） （KD group）,negative control group （NC group） and blank control group（CON group）. The infection efficiency of lentivirus for U937 was detected by flow cytometry. The mRNA and protein expression levels of HOXA10 were respectively detected by Real-Time PCR and Western Blot. Cell proliferation and apoptosis rates were de- tected by MTT and flow cytometry. RESULTS： The ratio of GFP positive cells was up to 90%. The mRNA and protein expression levels of HOXA10 were respectively （0. 047 3±0. 016 8） and （0. 001 5±0. 000 8） in KD group,and they were significantly lower than those in NC group （0. 225 2±0. 059 5,0. 013 4±0. 005 9,P values were 0. 004 and 0. 001） and in CON group （0. 261 3±0.054 9,0. 014 5±0. 001 6,P values were 0. 002 and 0. 001）. The IC50 of Ara-c and DNR in KD group [（12.80±4. 32）μg/mL, （75.34±20.40） ng/mL] were significantly lower than those in NC group [（116.65± 16.66） μg/mL,（245.00±46. 13） ng/mL,P values were less than 0.001 and 0.001] and in CON group [（100. 14±15.45） μg/mL,（267.67±32.65） ng/mL,both P〈0. 001]. The apoptosis rates induced by Ara-c and DNR in KD group [（71. 37±.24）%,（59.72±3.07）%] were significantly higher than those in NC group [（45.27± 1.88）%, （34.96±5.89）%,P values were less than 0. 001 and 0. 004] and in CON group [（39.39±4.46）%, （35.05±9.23）%P values were less than 0. 001 and 0. 004]. There were no significant differences at ICs0 and apoptosis rates between NC and CON group. CONCLUSION： Silencing HOXA10 gene may increase the sensitivity of U937 cells to chemotherapeutic drugs.

关 键 词：白血病 HOXA10慢病毒 RNA干扰 阿糖胞苷 柔红霉素 

分 类 号：R733.7[医药卫生—肿瘤]