MTX和IFN-α-2b诱导K562细胞凋亡及p73基因表达的研究  

Study on apoptosis of K562 cell line and expression of p73 gene induced by MTX and IFN-α-2b

作　　者：双跃荣[1] 樊光华[1] 

机构地区：[1]江西省肿瘤医院,江西南昌330029

出　　处：《肿瘤防治杂志》2004年第6期589-592,共4页China Journal of Cancer Prevention and Treatment

摘　　要：目的 :研究甲氨蝶呤 (MTX)和干扰素 α 2b(IFN α 2b)诱导人慢性粒细胞白血病急变细胞株 (K5 62 )凋亡的作用及凋亡相关基因p73mRNA在凋亡前后表达的变化。方法 :采用形态学观察法、DNA琼脂糖凝胶电泳和流式细胞仪检测K5 62细胞凋亡 ,采用半定量逆转录PCR方法 (RT PCR)检测p73mRNA的表达。结果 :4μg/mLMTX和60 0IU/mLIFN α 2b分别作用于K5 62细胞 6、12和 2 4h ,通过Wright’s +Giemsa染色可见部分细胞染色质明显固缩。DNA琼脂糖凝胶电泳可见清晰的梯形条带。流式细胞仪检测 ,MTX作用细胞6、12和 2 4h ,细胞凋亡率 (AR)分别为 5 15 %、14 70 %和 3 5 49% ;而IFN α 2b作用 6、12和 2 4h ,AR分别为 2 3 3 %、11 90 %和 3 5 15 %。对照组AR分别为 0 93 %和 1 0 9%。半定量RT PCR测定表明 ,IFN α 2b作用 2 4h组p73mRNA表达较对照组下调 ( 0 43± 0 0 5vs 0 66± 0 11,P <0 0 5 ) ,其余各组无明显变化。结论 :MTX、IFN α 2b可诱导K5 62细胞凋亡 ;在MTX、IFN α 2b诱导K5 62细胞凋亡的早期 ,p73mRNA表达无明显变化 ;IFN α 2b作用 2 4h ,p73mRNA表达下调。OBJECTIVE:To explore the variation of p73 gene expression in the apoptotic process of K562 cell line induced by methotrexate (MTX) and interferon-alpha-2b (IFN-α-2b).METHODS:Morphological changes of apoptotic cells were determined with Wright-Giemsa staining.DNA ladder and apoptotic cells were observed by the DNA agarose gel electronphoresis and flow cytometry (FCM) respectively.The expression of p73 mRNA was detected by the semi-quantitive reverse transcription polymerase chain reaction (RT-PCR).RESULTS:Apoptosis of K562 cells was induced by exposure to 4 μg/mL MTX or 600 IU/mL IFN-α-2b for 6,12,24 hours respectively.Condense of nucleus and DNA ladder was seen after the treatment of MTX and IFN-α-2b.Sub-G1 peak was also found by FCM.The expression of p73 mRNA was generally constant,except for the group treated with IFN-α-2b for 24 hours (0.43±0.05 vs 0.66±0.11,P<0.05).CONCLUSIONS:Both MTX and IFN-α-2b are capable of inducing apoptosis on K562 cells.p73 mRNA is detectable by semi-quantitive RT-PCR.The quantity of p73 mRNA is not changed in apoptotic K562 cells induced by MTX or IFN-α-2b,but when K562 cells are incubated with IFN-α-2b for 24 hours,p73 mRNA is regulated down.

关 键 词：P73基因 K562细胞 甲氨蝶呤 干扰素-Α-2B 

分 类 号：R733.72[医药卫生—肿瘤]