Atractylenolide I modulates ovarian cancer cell-mediated immunosuppression by blocking MD-2/TLR4 complex-mediated MyD88/NF-κB signaling in vitro.
Atractylenolide I modulates ovarian cancer cell-mediated immunosuppression by blocking MD-2/TLR4 complex-mediated MyD88/NF-κB signaling in vitro.
复制标题
苍术素 I 在体外通过阻断 MD - 2/TLR4 复合物介导的 MyD88/NF - κB 信号通路，调节卵巢癌细胞介导的免疫抑制。
DOI：
10.1186/s12967-016-0845-5
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发表时间：
2016-04-27
影响因子：
7.4
通讯作者：
Huang W
中科院分区：
文献类型：
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作者：
Liu H;Zhang G;Huang J;Ma S;Mi K;Cheng J;Zhu Y;Zha X;Huang W
TLR 4/MD-2复合物介导的NF-κB和Akt的MyD 88依赖性活化通过诱导免疫抑制细胞因子和吲哚胺2，3-双加氧酶（IDO）促进上皮性卵巢癌（EOC）的肿瘤相关免疫抑制青蒿素I（AO-1）是一种天然存在的倍半萜内酯，已知可改变EOC细胞上人MD-2的构象系综。本研究探讨AO-1对TLR 4/MD-2复合物介导的MyD 88/NF-κB信号通路的调节作用。Western blot检测NF-κB、Akt和IDO 1在MyD 88 + EOC SKOV 3细胞中的表达和活化;流式细胞术检测SKOV 3细胞表面TLR 4/MD-2复合物和T淋巴细胞表型; L-犬尿氨酸检测IDO活性; ELISA检测免疫抑制细胞因子; MTT法检测T细胞对丝裂原刺激的增殖;采用LDH-细胞毒性试验测定淋巴细胞和NK细胞的细胞毒性。AO-1可下调TLR 4/MD-2复合物的表达，从而下调MyD 88/NF-κB信号通路，下调NF-κB、Akt和IDO 1的激活，下调IL-6、TGF-β1、VEGF和IL-17 A的分泌，并进一步降低调节性T细胞的水平（Treg细胞）并改善暴露于EOC SKOV 3细胞上清液的T淋巴细胞的降低的增殖反应和抗肿瘤细胞毒性。AO-1可能通过阻断TLR 4/MD-2复合物介导的MyD 88/NF-κB信号通路逆转EOC细胞介导的免疫抑制。
TLR4/MD-2 complex-mediated MyD88-dependent activation of NF-κB and Akt promotes tumor-associated immunosuppression in epithelial ovarian cancer (EOC) via induction of immunesuppressive cytokines and indoleamine 2,3-dioxygenase (IDO). Atractylenolide I (AO-1) is a naturally occurring sesquiterpene lactone known to change the conformational ensemble of human MD-2 on EOC cells. This study examined the modulation by AO-1 of TLR4/MD-2 complex-mediated MyD88/NF-κB signaling. The expression and activation of NF-κB, Akt and IDO1 by MyD88+ EOC SKOV3 cells was determined using western blot; the TLR4/MD-2 complex on SKOV3 cells and the phenotype of T lymphocytes were determined using flow cytometry; IDO activity was evaluated by measuring l-kynurenine; Immunesuppressive cytokines were detected using ELISA; T‐cell proliferation to mitogen stimulation was assessed by MTT assay; the cytotoxicity of lymphocytes and NK cells was measured using LDH-cytotoxicity assay. AO-1 could down-regulate expression of TLR4/MD-2 complex, resulting in downregulation of MyD88/NF-κB signaling and activation of NF-κB, Akt and IDO1 and secretion of IL-6, TGF-β1, VEGF and IL-17A by EOC SKOV3 cells, and further reduce increased levels of regulatory T cells (Treg cells) and improve decreased proliferative response and antitumor cytotoxicity of T lymphocytes exposed to EOC SKOV3 cell supernatant. AO-1 may reverse EOC cell-mediated immunosuppression through blocking TLR4/MD-2 complex-mediated MyD88/NF-κB signaling.