Polyphenols from marine brown algae target radiotherapy-coordinated EMT and stemness-maintenance in residual pancreatic cancer.
Polyphenols from marine brown algae target radiotherapy-coordinated EMT and stemness-maintenance in residual pancreatic cancer.
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来自海洋褐藻的多酚针对残余胰腺癌中放射治疗协调的 EMT 和干细胞维持。
DOI：
10.1186/s13287-015-0173-3
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发表时间：
2015-09-22
影响因子：
7.5
通讯作者：
Aravindan N
中科院分区：
文献类型：
--
作者：
Aravindan S;Ramraj SK;Somasundaram ST;Herman TS;Aravindan N
在存活的肿瘤细胞中，与治疗相关的干细胞维持作用决定了肿瘤的复发/复发。最近，我们认识到海藻多酚复合物具有抗胰腺癌（PC）的潜力，并缩小了三种可作为胰腺癌治疗佐剂的优良药物交付物。利用离体培养的PC-癌干细胞（PC- cscs）和残余PC小鼠模型，我们研究了海藻多酚在调节干细胞维持中的作用。将Panc-1、Panc-3.27、MiaPaCa-2或BxPC-3细胞来源的异种移植物的ALDH+CD44+CD24+ PC-CSCs分别进行模拟辐照、分次辐照（FIR, 2Gy/D） 5天，再用带/不带FIR的三聚体（HT-EA）、asperum （SA-EA）或Padina tetrastromatica （PT-EA）多酚（100 μg/ml）处理，检测细胞活力和93个干细胞相关分子的转录（QPCR分析）。在Panc-1/Panc-3.27/MiaPaCa-2/ bxpc -3异种移植物衍生的PC-CSCs中检测fir1转激活的Oct4、Zic3、EIF4C、Nanog和LIF的多酚依赖性调节（QPCR）以及Nanog、SOX2和OCT3/4的功能翻译（免疫印迹法）。采用组织芯片构建和自动免疫组化技术检测海藻多物质在治疗（FIR, 2Gy/D， 5D/周，持续3周）耐药残余肿瘤中对EMT （N-Cadherin）、多能性（SOX2, OCT3/4, Nanog）和干细胞维持（PI3KR1， LIF, CD44）的调控作用。体外暴露于SA-EA、PT-EA和HT-EA的PC-CSCs表现出剂量依赖性的细胞活力抑制。FIR扩增了MiaPaCa-2-、Panc-1-、Panc-3.27-和bxpc3构建的异种移植来源的ALDH+CD44+CD24+PC-CSCs中69、80、74和77个干细胞相关基因的转录。SA-EA、PT-EA或HT-EA处理完全抑制了由MiaPaCa-2、Panc-1、Panc-3.27和BXPC3异种移植物建立的ALDH+CD44+CD24+PC-CSCs中fir激活的干细胞转录机制。QPCR验证了EIF4C、OCT3/4、Nanog、LIF和ZIC3转录谱结果。Nanog、Sox2和OCT3/4免疫印迹证实了海藻多酚对PC-CSC的放射增敏作用。经体内处理后的残余pc组织微阵列和免疫染色证实，SA-EA、PT-EA和HT-EA完全调节了fir诱导的SOX2、OCT3/4、Nanog、LIF、CD44、PIK3R1、N-Cadherin和E-Cadherin。这些数据首次记录了耐药PC-CSCs的EMT/干细胞维持情况。此外，数据表明，海藻多酚可能通过靶向残余细胞中治疗精心策划的干细胞信号传导来抑制PC复发/复发。本文的在线版本（doi:10.1186/s13287-015-0173-3）包含补充材料，仅供授权用户使用。
Therapy-associated onset of stemness-maintenance in surviving tumor-cells dictates tumor relapse/recurrence. Recently, we recognized the anti-pancreatic cancer (PC) potential of seaweed polyphenol manifolds and narrowed down three superior drug-deliverables that could serve as adjuvants and benefit PC cure. Utilizing the PC- cancer stem cells (PC-CSCs) grown ex vivo and mouse model of residual-PC, we investigated the benefits of seaweed polyphenols in regulating stemness-maintenance. ALDH+CD44+CD24+ PC-CSCs from Panc-1, Panc-3.27, MiaPaCa-2, or BxPC-3 cells-derived xenografts grown ex vivo were either mock-irradiated, exposed to fractionated irradiation (FIR, 2Gy/D for 5 days), treated with polyphenols (100 μg/ml) of Hormophysa triquerta (HT-EA), Spatoglossum asperum (SA-EA) or Padina tetrastromatica (PT-EA) with/without FIR were examined for cell viability, transcription of 93 stem-cell-related molecules (QPCR profiling). Polyphenol-dependent regulation of FIR-transactivated Oct4, Zic3, EIF4C, Nanog, and LIF (QPCR) and functional translation of Nanog, SOX2, and OCT3/4 (immunoblotting) were examined in Panc-1/Panc-3.27/MiaPaCa-2/BxPC-3-xenografts derived PC-CSCs. Effect of seaweed-polyphenols in the regulation of EMT (N-Cadherin), pluripotency- (SOX2, OCT3/4, Nanog) and stemness-maintenance (PI3KR1, LIF, CD44) in therapy (FIR, 2Gy/D for 5D/wk for 3-weeks) resistant residual tumors were examined by tissue microarray construction and automated immunohistochemistry. Ex vivo exposure of PC-CSCs to SA-EA, PT-EA and HT-EA exhibit dose-dependent inhibition of cell viability. FIR amplified the transcription of 69, 80, 74 and 77 stem-cell related genes in MiaPaCa-2-, Panc-1-, Panc-3.27- and BXPC3-established xenograft-derived ALDH+CD44+CD24+PC-CSCs. Treatment with SA-EA, PT-EA, or HT-EA completely suppressed FIR-activated stem-cell transcriptional machinery in ALDH+CD44+CD24+PC-CSCs established from MiaPaCa-2, Panc-1, Panc-3.27 and BXPC3 xenografts. QPCR validated EIF4C, OCT3/4, Nanog, LIF, and ZIC3 transcriptional profile outcomes. Nanog, Sox2, and OCT3/4 immunoblotting affirmed the PC-CSC radiosensitizing benefit of seaweed polyphenols. Residual-PC tissues microarrayed and immunostained after in vivo treatments recognized complete regulation of FIR-induced SOX2, OCT3/4, Nanog, LIF, CD44, PIK3R1, N-Cadherin, and E-Cadherin with SA-EA, PT-EA, and HT-EA. These data, for the first time, documented the EMT/stemness-maintenance in therapy-resistant PC-CSCs. Further, the data suggest that seaweed polyphenols may inhibit PC relapse/recurrence by targeting therapy-orchestrated stem-cell signaling in residual cells. The online version of this article (doi:10.1186/s13287-015-0173-3) contains supplementary material, which is available to authorized users.