吉西他滨诱导人NK/T细胞淋巴瘤细胞株SNK-6凋亡的机制

目的探讨吉西他滨诱导人NK/T细胞淋巴瘤细胞株SNK-6产生凋亡的机制。方法应用MTT法、DNA电泳技术及流式细胞术观察吉西他滨诱导人SNK-6细胞凋亡情况，应用基因芯片技术分析加药前后凋亡相关基因的表达差异。结果吉西他滨作用后，MTT结果显示细胞生长抑制率随药物浓度的增加而逐渐增大；DNA断裂电泳可见典型的梯形DNA条带，随吉西他滨作用浓度的增加而逐渐增强；流式细胞术分析，2 μg/ml吉西他滨作用SNK-6细胞4、8、24、48 h后，凋亡细胞比例分别为2.1%、8.3%、23.9%、30.9%；吉西他滨作用48 h后，在88条有关凋亡的目的基因中共有17条基因表达发生大于3倍的显著变化,其中表达上调基因4条，下调基因13条。结论吉西他滨可以诱导人NK/T细胞淋巴瘤细胞产生凋亡，呈时间和剂量依赖性；吉西他滨可使SNK-6细胞中多个凋亡相关基因的表达发生改变，初步揭示了其诱导淋巴瘤细胞凋亡的作用机制。

Apoptosis of NK/T Lymphoma Cell Line SNK-6 Induced by Gemcitabine

ObjectiveTo investigate the apoptosis induced by gemcitabine in human NK/T lymphoma cell line SNK-6.MethodsMTT, DNA fragmentation and flow cytometry (FCM) analysis were used to detect apoptosis of SNK-6 cells induced by gemcitabine. A human apoptosis microarray containing 88 cDNA fragments was used to detect apoptosis related genes expression difference.ResultsGemcitabine could inhibit the growth rate of SNK-6 cells after 48h treatment in a concentration-dependent manner. DNA ladder was observed on DNA electrophoresis and its intensity increased with the increased concentration of gemcitabine. FCM assay revealed that apoptosis cells respectively accounted for 2.1%,8.3%,23.9%,30.9% of total cells respectively after exposure to 2 μg/ml gemcitabine for 4, 8, 24, 48 h. cDNA microarray analysis identified 4 up-regulated and 13 down-regulated genes with more than three-fold change in the first 48h as a consequence of treatment.ConclusionGemcitabine can induce apoptosis of human NK/T lymphoma cell line SNK-6 cells in a time-dependent and concentration-dependent manner and alter the expression profile of apoptosis-related genes in this cell line, which provides valuable insight into the mechanism of the gemcitabine-induced apoptosis.