| 平超强 张宇晶 孙国勋 董敏 周鑫磊 平家奇 刘晔 洪珞珈.DC-CIK对K562/A多药耐药基因mdr1 表达的影响[J].,2014,14(14):2667-2671 |
| DC-CIK对K562/A多药耐药基因mdr1 表达的影响 |
| The Expression Effect of DC-CIK on K562/A Multidrug ResistanceGene mdr1 |
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| DOI： |
| 中文关键词: 树突状细胞（DC） 细胞因子诱导的杀伤细胞（CIK） K562/A mdr1 逆转耐药 |
| 英文关键词: Dendritic cells Cytokine-induced killer cells K562/A Mdr1 Reversal of drug resistance |
| 基金项目:黑龙江省留学归国人员基金（LC08C24）；黑龙江省科技厅课题（QC2012C050） |
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| 中文摘要: |
| 目的：体外观察树突状细胞（dendritic cell，DC）联合细胞因子诱导的杀伤细胞（cytokine inducedkiller，CIK）对K562/A细胞株
多药耐药基因mdr1 表达的影响。方法：采集健康人的外周血，分离出单个核细胞( peripheral blood mononuclear cell，PBMC )，在
体外加入多种细胞因子经诱导生成DC及CIK 细胞，以流式细胞仪检测其表面标志，将DC 细胞内加入K562/A 细胞裂解物致敏
后，再与CIK细胞混合培养48 小时。将致敏后的DC-CIK 细胞与K562/A及K562 分组培养后以荧光定量PCR 检测其mdr1 基
因表达的情况，PBMC 作为对照组。结果：RT-PCR 中可见K562/A+DC-CIK 组中mdr1 mRNA 表达较K562/A明显降低，经荧光定
量PCR 观察到K562/A 内mdr1 mRNA 表达为K562 的10.27 倍、K562/A/PBMC 略低于未处理的K562/A（P>0.05），
K562/A/DC-CIK 细胞中mdr1 mRNA含量较K562/A、K562/A/PBMC 少（P<0.05）。DC-CIK细胞与细胞株混合培养后，mdr1 基因
表达较混合培养前明显降低。结论：实验数据显示DC-CIK 可使耐药细胞株内mdr1 基因表达下调。但K562 与DC-CIK 混合培养
后该基因降低不明显，提示该基因在细胞中存在着基础表达，意义在于维持细胞内稳态。目前针对逆转白血病耐药的研究较少，
需要多进行相关研究以拓宽细胞免疫治疗在逆转耐药领域的应用。DC-CIK 是具有发展潜力的抗肿瘤方法。本实验将为下一阶段
研究逆转耐药的机制提供依据，DC-CIK 细胞免疫疗法有望成为逆转肿瘤耐药的新方法。 |
| 英文摘要: |
| Objective:Observe the expression effect of dendritic cells (DC) combined with cytokine-induced killer cells (CIK) on
the multidrug resistance gene mdr1 of K562/A cell line in vitro.Methods:Peripheral blood mononuclear cells (PBMC) isolated from
peripheral blood of healthy people were collected, and induced them to DC and CIK by mixing in a variety of cytokines,then detected
their surface markers by flow cytometry. Added cell lysates of K562/A to DC, and then made the sensitized DC culturing with CIK for 48
hours. Detected the expression of mdr1 gene by Quantitative PCR after sensitized DC-CIK were cultured with K562/A and K562
respectively, used PBMC as a control group.Results:The mdr1 mRNA in K562 / A + DC-CIK group was significantly lower than K562 /
A group by RT-PCR. Through Quantitative PCR we observed that the expression of mdr1 mRNA in K562 / A group was 10.27 times
than K562 group, K562/A/PBMC group was slightly lower than the untreated K562/A group（P>0.05）, and K562/A/DC-CIK group was
lower than K562/A、K562/A/PBMC group （P<0.05）. The expression of mdr1 gene after mixed culture was significantly lower than
DC-CIK mixed culture with two cell lines before.Conclusion:The experimental datas showed that mdr1 gene in resistant cell lines can
be downregulated by DC-CIK. But this gene was not reduced significantly after K562 mixed culture with DC-CIK, and it suggesting that
there is basal gene expression in cells to maintain the intracellular homeostasis. The research about reversing resistance against leukemia
is few currently, need to conduct more research in order to broaden the immunotherapy field of application in reversing drug resistance.
DC-CIK is a anti-tumor approach with development potential. This experiment will provide a basis for the study of reversing drug
resistance mechanisms next phase , and DC-CIK cellular immunotherapy is expected to become a new way reversing tumor resistance.
The mothod is expected to become the leader in cancer treatment, it has broad prospects for development. |
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