Downregulation of miR-302c and miR-520c by 1,25(OH)2D3 treatment enhances the susceptibility of tumour cells to natural killer cell-mediated cytotoxicity.
Downregulation of miR-302c and miR-520c by 1,25(OH)2D3 treatment enhances the susceptibility of tumour cells to natural killer cell-mediated cytotoxicity.
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1,25(OH)2D3 治疗下调 miR-302c 和 miR-520c 增强肿瘤细胞对自然杀伤细胞介导的细胞毒性的敏感性
DOI：
10.1038/bjc.2013.337
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发表时间：
2013-08-06
影响因子：
8.8
通讯作者：
Hu H
中科院分区：
文献类型：
--
作者：
Min D;Lv XB;Wang X;Zhang B;Meng W;Yu F;Hu H
背景：NKG2D识别多种配体，包括多态性主要组织相容性复合体I类链相关链相关蛋白A和B （MICA/B）和独特的长16结合蛋白（ulbp）。这些配体存在于癌细胞上，并被NKG2D以细胞结构感应的方式识别，触发自然杀伤（NK）细胞毒性。然而，恶性细胞中控制NKG2D配体表达的机制尚不清楚。1-α，25-二羟基维生素D3 （1,25(OH)2D3）最近被证明可以增强黑色素瘤细胞对NK细胞毒性的敏感性。然而，1,25(OH)2D3在其他癌症中的功能及其潜在的作用机制尚不清楚。方法：采用实时荧光定量PCR检测miR-302c和miR-520c在Kasumi-1、K562、MCF7和MDA-MB-231细胞中的表达水平。通过荧光素酶报告基因检测确认miR-302c和miR-520c的靶点。采用CytoTox 96非放射性细胞毒法检测NK92细胞对Kasumi-1、K562、MCF7和MDA-MB-231细胞的杀伤作用。用酶联免疫吸附法测定NK细胞活化的细胞因子IFN-γ和颗粒酶B的水平。结果：1,25(OH)2D3增强血液学肿瘤细胞系Kasumi-1和实体瘤细胞系MDA-MB-231对NK92细胞的敏感性。miR-302c和miR-520c被诱导表达，在1,25(OH)2D3处理下，它们的水平与NKG2D配体MICA/B和ULBP2的水平呈负相关。荧光素酶报告基因分析显示，miR-302c和miR-520c直接靶向MICA/B和ULBP2的3 ' - utr，负调控MIA/B和ULBP2的表达。此外，通过转染miR-302c或miR-520c的模拟物来上调miR-302c或miR-520c，可显著降低NK细胞共孵育后Kasumi-1细胞的活力。相反，通过各自的反义寡核苷酸抑制miR-302c或miR-520c的活性可以提高Kasumi-1细胞对NK细胞的抗性。结论：1,25(OH)2D3促进NK细胞对恶性细胞的免疫攻击，部分是通过下调miR-302c和miR-520c，从而上调NKG2D配体MICA/B和ULBP2。
Background:NKG2D recognises several ligands, including polymorphic major histocompatibility complex class I chain-related chain-related proteins A and B (MICA/B) and unique long 16-binding proteins (ULBPs). These ligands are present on cancer cells and are recognised by NKG2D in a cell-structure-sensing manner, triggering natural killer (NK) cell cytotoxicity. However, the mechanisms that control the expression of NKG2D ligands in malignant cells are poorly understood. 1-α,25-Dihydroxyvitamin D3 (1,25(OH)2D3) was recently shown to enhance the susceptibility of melanoma cells to the cytotoxicity of NK cells. However, the function of 1,25(OH)2D3 in other cancers and its potential mechanisms of action remain unknown.Methods:The expression levels of miR-302c and miR-520c in Kasumi-1, K562, MCF7 and MDA-MB-231 cells were evaluated using quantitative real-time PCR. The targets of miR-302c and miR-520c were confirmed by luciferase reporter assay. The killing effects of NK92 cells against Kasumi-1, K562, MCF7 and MDA-MB-231 cells were examined using the CytoTox 96 Non-Radioactive Cytotoxicity Assay. The levels of cytokines IFN-γ and granzyme B, which indicate the activation of NK cells, were also measured by enzyme-linked immunosorbent assay.Results:Treatment with 1,25(OH)2D3 enhanced the susceptibility of both the haematological tumour cell line Kasumi-1 and solid tumour cell line MDA-MB-231 to NK92 cells. miR-302c and miR-520c expression was induced, and their levels inversely correlated with the levels of NKG2D ligands MICA/B and ULBP2 upon 1,25(OH)2D3 treatment. A luciferase reporter assay revealed that miR-302c and miR-520c directly targeted the 3′-UTRs of MICA/B and ULBP2 and negatively regulated the expression of MIA/B and ULBP2. Moreover, upregulation of miR-302c or miR-520c by transfection of their mimics remarkably reduced the viability of Kasumi-1 cells upon NK cell co-incubation. By contrast, the suppression of the activity of miR-302c or miR-520c by their respective antisense oligonucleotides improved the resistance of Kasumi-1 cells to NK cells.Conclusion:1,25(OH)2D3 facilitates the immuno-attack of NK cells against malignant cells partly through downregulation of miR-302c and miR-520c and hence upregulation of the NKG2D ligands MICA/B and ULBP2.
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DOI：
10.1097/ppo.0b013e3181c51ee6
发表时间：
2010-01
期刊：
Cancer journal (Sudbury, Mass.)
影响因子：
--
作者：
Trump DL;Deeb KK;Johnson CS
通讯作者：
Johnson CS
影响因子：
3.5
作者：
Kizildag, Sefa;Ates, Halil;Kizildag, Servet
通讯作者：
Kizildag, Servet
影响因子：
20.3
作者：
Trotta, Rossana;Chen, Li;Caligiuri, Michael A.
通讯作者：
Caligiuri, Michael A.
DOI：
10.1073/pnas.201238598
发表时间：
2001-09-25
影响因子：
11.1
作者：
Cerwenka, A;Baron, JL;Lanier, LL
通讯作者：
Lanier, LL
影响因子：
30.5
作者：
Eisenring, Maya;vom Berg, Johannes;Becher, Burkhard
通讯作者：
Becher, Burkhard