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结外NK/T细胞淋巴瘤敏感药物筛选

Chinese Journal of Cancer Prevention and Treatment(2015)

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Abstract
目的 应用无血清培养基(serum free medium,SFM)初步富集肿瘤耐药细胞,筛选出结外NK/T细胞淋巴瘤(extranodal NK/T-celllymphoma,ENKTCL)的敏感药物.方法 应用加入10%人血清和700 U/mL人白细胞介素-2(interlukin-2,IL-2)的1640培养基和加入700 U/mL人IL-2的SFM VIVO-15TM,分别培养SNK-6细胞;MTT法分别检测2种培养体系中表柔比星(adriamycin,ADM)、奥沙利铂(oxaliplatin,L-OHP)、吉西他滨(gemcitabine,GEM)、左旋门冬酰胺酶(L-Asparaginase,L-ASP)、阿糖胞苷(cytosine arabinoside,Ara-C)和甲氨蝶呤(methotrexate,MTX)等药物作用的IC50;2种培养体系中分别加入上述各种药物作用32 h后,7AAD/PE-Annexin Ⅴ染色法流式细胞术检测各处理组细胞的凋亡率,并分析比较.结果 无血清培养体系中细胞部分悬浮生长,每个悬浮球由>50个数目不等的细胞组成,悬浮球体积较有血清中明显增大.有血清和SFM中IC50含量,ADM分别为(0.12±0.018)和(0.751±0.14) μg/mL,P<0.001;Ara-C分别为(0.217±0.002)和(0.822±0.028) μg/mL,P<0.001; MTX分别为(0.023±0.001)和(0.032±0.002) μg/mL,P=0.002.药物作用后,有血清和SFM中SNK-6细胞的凋亡率,ADM分别为(45.23±2.58)%和(30.91±2.13)%,P=0.041;Ara-C分别为(56.12±2.32)%和(41.47±2.46)%,P=0.034;MTX分别为(24.42±1.92)%和(13.01±2.28)%,P=0.045;GEM分别为(15.05±2.05)%和(41.45±1.74)%,P<0.001.结论 SFM VIVO-15TM可用于ENKTCL SNK-6细胞培养;无血清培养后的SNK-6细胞对ADM、Ara-C和MTX耐药性增强,对L-OHP、GEM和L-ASP敏感.
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Key words
MTT,apoptosis,serum free medium,SFM,tumor-initiating cells
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