磁性纳米铁联合多柔比星对Raji细胞的影响

 目的　观察干扰素α（IFN-α）对恶性造血细胞系主要组织相容性复合体（MHC）Ⅰ类链相关蛋白A（MICA）表达的影响。方法　反转录-聚合酶链反应（RT-PCR）法检测肿瘤细胞MICA mRNA的表达，免疫组织化学法检测MICA蛋白的表达，四甲基偶氮唑蓝（MTT）比色法检测人类外周血自然杀伤（NK）细胞对肿瘤细胞的杀伤。结果　人类慢性髓系白血病细胞K562 MICA表达阳性，人类伯基特淋巴瘤细胞Raji MICA表达阴性。Raji和K562细胞分别在1000 U／ml IFN-α诱导24 h（t＝17.016，P＜0.05）和48 h（t＝5.616，P＜0.05）时MICA mRNA开始表达上调。72 h Raji细胞和K562细胞分别在500 U／ml（t＝6.622，P＜0.05）和1000 U／ml（t＝9.071，P＜0.05）诱导条件下MICA mRNA表达开始上调。IFN-α对MICA mRNA表达的上调作用呈一定时间和剂量依赖关系，且蛋白与mRNA表达水平一致。1000 U／ml IFN-α诱导72 h后，Raji细胞（t＝20.016，P＜0.05）和K562细胞（t＝7.969，P＜0.05）对NK细胞杀伤的敏感性明显上调，抗MICA抗体封闭MICA后，NK细胞对Raji细胞杀伤率恢复至诱导前水平（t＝0.393，P＞0.05），而K562细胞则低于诱导前水平（t＝9.841，P＜0.05）。结论　IFN-α上调了恶性造血细胞中MICA 基因的转录表达，继而增强了NK细胞对其识别与杀伤。

Effect of combination with magnetic nanoparticle of Fe3O4 and adriamycin on Raji cell line

Objective To investigate the effect of combination with magnetic nanoparticle of Fe3O4and adriamycin (ADM) on Raji cell line.Methods Raji cells were cultured with Fe3O4-magnetic nanoparticle and ADM after using mechanical absortion polymerization,and the cell viability was detected by MTT and trypan-blue exclusion.The apoptosis was detected by flow cytometry,and the expression of p53 and NF-κB were measured by Western blot. Results Raji cell proliferation ratio was significantly inhibited in concentration-and time-dependent manners in both ADM and Fe3O4-MNP-ADM groups (r =0.412,P =0.027;r =0.523,P =0.014).The percentages of apoptotic cells induced by ADM and Fe3O4-MNP-ADM were 8.76 ％ vs 14.85 ％,35.08 ％ vs 44.50 ％,44.00 ％ vs 69.40 ％ in 12,24 and 48 hours,respectively (t =-9.137,-4.808,-6.337; P =0.012,0.041,0.024).The grey straps of NF-κB measured by Western blot were 4.22±0.32,3.31±0.28 in ADM and Fe3O4-MNP-ADM group (t =-54.416,P =0.035),whereas the grey straps of p53 were 1.042±0.114,1.270±0.091,respectively (t =33.963,P =0.047).Conclusion Combination therapy of Fe3O4-MNP and ADM could inhibit proliferation and induce apoptosis of Raji cells line,which may due to upregulation of p53 and down-regulation of NF-κB.