AIM: To investigate the effect of focal adhesion kinase ( FAK ) gene silencing on chemotherapeutic drug sensitivity in leukemic cells.METHODS: Lentiviral - FAK - shRNA was transfected into BCR/ABL - BaF3 leukemic cells.The protein expression of FAK was detected by Western blotting.BCR/ABL - BaF3 leukemic cells were treated with different concentrations of imatinib in vitro, and the apoptosis was determined by labeling with Annexin V.A murine model of leukemia was established and the mice were treated with FAK shRNA and imatinib.Survival time and distribution of leukemic cells in bone marrow and spleen of the mice were monitored.RESULTS: FAK shRNA was successfully constructed and effectively inhibited FAK gene expression.With 5 μmol/L imatinib treatment, the percentages of apoptotic cells in vector control group and FAK shRNA group were ( 9.76 ± 1.97 )％ and ( 21.90 ±3.20 )％ , respectively, and significant difference between these 2 groups ( P ＜ 0.05 ) was observed.With 50 μmol/L imatinib treatment, the percentages of apoptotic cells in vector control group and FAK shRNA group were ( 56.10 ± 6.00 )％ and ( 82.10 ± 5.70 )％ , respectively, also with significant difference between these 2 groups ( P ＜0.05 ).Compared with vector control group, the mice in FAK gene silencing group displayed significantly prolonged survival time.Moreover, 60 days after injection of leukemic cells, the percentages of leukemic cells in bone marrow and spleen of the mice were significantly decreased in FAK gene silencing group as compared with those in vector control group.CONCLUSION: FAK gene silencing promotes the efficacy of chemotherapeutic drug in leukemic cells, indicating that FAK gene silencing might be considered as a new therapeutic strategy for leukemia.%目的:本实验研究黏着斑激酶(focal adhesion kinase,FAK)基因沉默对白血病细胞化疗药物敏感性的影响.方法:构建FAK shRNA慢病毒载体,转染至BCR/ABL-BaF3白血病细胞株,通过蛋白质印迹方法检测FAK蛋白表达情况.体外应用不同浓度的化疗药物伊马替尼处理BCR/ABL-BaF3细胞,予Annexin V标记检测细胞凋亡情况.建立白血病动物模型,联合应用FAK shRNA及伊马替尼进行处理,观察小鼠生存情况,分析白血病细胞在小鼠骨髓及脾脏的分布情况.结果:成功建立FAK shRNA慢病毒载体,该载体能有效沉默FAK基因表达.体外实验发现5 μmol/L 伊马替尼处理时,空白对照组与FAK基因沉默组中Annexin V阳性细胞百分比分别为(9.76±1.97)%和(21.90±3.20)%,差异显著(P<0.05);予50 μmol/L 伊马替尼处理时,2组中Annexin V阳性细胞百分比分别为(56.10±6.00)%和(82.10±5.70)%,差异显著(P<0.05).白血病动物实验中,与空白对照组相比,生存分析结果显示FAK基因沉默组小鼠的生存时间明显延长.白血病细胞输注后第60 d,与空白对照组相比,白血病细胞在FAK基因沉默组小鼠骨髓和脾脏的分布明显减少.结论:FAK基因沉默能明显增强白血病细胞对化疗药物敏感性,提示FAK基因沉默有望成为白血病治疗的新策略.
展开▼
