新型bcl-2反义寡核苷酸F951对人白血病裸鼠移植瘤的治疗作用  

Effect of F951,a novel bcl-2 antisense oligodeoxynucleotide,on human leukemia transplanted subcutaneously in nude mice

作　　者：李东良[1,2] 吕联煌[1] 林锦娟[1] 林振兴[1] 陈英玉[1] 

机构地区：[1]福建医科大学附属协和医院血液病研究所,福建福州350001 [2]南京军区福州总医院肝胆病中心,福建福州350025

出　　处：《中国药理学通报》2009年第2期230-234,共5页Chinese Pharmacological Bulletin

基　　金：福建省“211”工程重点学科专项资助

摘　　要：目的研究新型反义寡核苷酸F951对急性髓性白血病(AML)裸鼠移植瘤bcl-2基因表达、肿瘤生长及荷瘤鼠生存期的影响。方法体外大量扩增高表达bcl-2基因的HL60细胞,皮下接种建立AML裸鼠移植瘤模型,采取瘤体局部注射给药,观察F951及F951联合小剂量Ara-C对肿瘤生长及荷瘤裸鼠生存期的影响;应用荧光定量RT-PCR检测瘤细胞bcl-2mRNA表达;光镜观察瘤组织形态结构变化。结果治疗14d各组荷瘤鼠瘤体积、瘤重、瘤组织bcl-2mR-NA定量分别为:NS对照组(15.17±3.40)cm3、(12.69±0.92)g、9.79×104Copies.μg-1;无义寡核苷酸(FNS)组(15.91±3.77)cm3,(12.38±1.21)g;8.31×104Copies.μg-1;Ara-C组(1.24±0.55)cm3,(2.32±0.49)g,2.59×104Copies.μg-1;F951组(2.6±1.55)cm3,(3.53±0.67)g;1.01×10Copies.μg-1;F951+Ara-C组(0.62±0.48)cm3,(1.05±0.63)g,9.5×102Copies.μg-1;上述各组数据显示F951有下调肿瘤细胞bcl-2基因表达,抑制肿瘤生长的作用,抑瘤率为72.18%,与NS及FNS对照组比差异均有极显著性(P<0.01),F951联合Ara-C后治疗效果更佳,抑瘤率达91.73%,与Ara-C组相比差异有显著性(P<0.05)。瘤组织病理学检查:F951及F951联合Ara-C治疗组瘤细胞减少,瘤细胞变性坏死,大量纤维组织增生。各组动物生存期观察:NS对照组(13.67±0.82)d、,FNS组(13.50±1.22)d,Ara-C组(28.33±1.86)d,F951组(29.33±7.44)d,F951+Ara-C组(37.83±5.85)d。F951组及F951+Ara-C组生存期延长率分别为114.56%与176.71%,明显延长了荷瘤鼠的生存期(P<0.01)。结论F951能特异性地抑制AML裸鼠移植瘤细胞bcl-2基因表达,激活凋亡调控机制,诱导瘤细胞凋亡,同时增加化疗药物的敏感性,而发挥抗肿瘤作用。Aim To study the inhibitory effects of F951, a novel bcl-2 antisense oligodeoxynucleotide, on expression of bcl-2 , growth of tumor and survival time of nude mice transplanted subcutaneously with acute myeloid leukemia. Methods HL-60 cells with high expression of bcl-2 were proliferated in vitro. The models of the nude mice with HL-60 cells were established by subcutaneous transplantation with drugs directly injected. The effects of F951 and F951 with low dose Ara-c on growth of tumor and survival time of mice with tumor were observed. The expressions of bcl-2 mRNA in the tumors implanted were detected by fluorescent quantitation RT-PCR. The morphologic structure of tumor tissues was assayed by light microscope. Results After each group mice with tumors were treated for 14 days, the volume,the weight of tumor and the bcl-2 mRNA expression of tumor tissue were shown respectively as follows： NS control group （15.174±3.40）cm^3、（12.694±0.92）g、9.79×10^4 Copies·μg^-1;FNS group （15.914±3.77）cm^3,（12.384±1.21）g;8.31×10^4Copies·μg^-1;Ara-C group（1.244±0.55）cm^3,（2.324±0.49）g,2.59×10.Copies·μg^-1;F951 group（2.64±1.55）cm^3,（3.534±0.67）g;1.01×10Copies·μg^-1;F951＋Ara-C group （0.624±0.48）cm^3,（1.054±0.63）g,9.5×10^2Copies·μg^-1 The data above showed that F951 could downregulate the expression of bcl-2 in nude mice with HL-60 cells xenograft and inhibit growth of tumor. The growth of tumor of F951 group was reduced, and the inhibitory rate was 72. 18% , there was significant difference comparing control groups with NS and FNS （P 〈0.01） . The growth of tumorof F951 with low-dose Ara-e group was reduced,and the inhibitor rate was 91.73%, there was significant difference compared with Ara-C groups（ P 〈 0.05） . The result of pathologi tissue showed the tumor cells i examination of tumor F951 and F951 with Ara-c groups had significant reduction, degeneration and necrosis and there were a lot of fibroplasia in tumor tissue. The survival time of mice

关 键 词：白血病 骨髓细胞 急性 基因 bcl-2 寡核苷酸类 反义 药效学 

分 类 号：R-332[医药卫生] R329.25