Host-Defense Peptides Caerin 1.1 and 1.9 Stimulate TNF-Alpha-Dependent Apoptotic Signals in Human Cervical Cancer HeLa Cells
Host-Defense Peptides Caerin 1.1 and 1.9 Stimulate TNF-Alpha-Dependent Apoptotic Signals in Human Cervical Cancer HeLa Cells
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DOI：
10.3389/fcell.2020.00676
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发表时间：
2020-07-31
影响因子：
5.5
通讯作者：
Wang, Tianfang
中科院分区：
文献类型：
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作者：
Ni, Guoying;Chen, Shu;Wang, Tianfang
从澳大利亚树蛙属（Litoria）的腺分泌物中分离的宿主防御肽Caerin 1.1和1.9具有多种生物学活性，包括抑制人乳头瘤病毒（HPV）16早期蛋白E7转化的小鼠以及人癌细胞在体外和体内的增殖。然而，其对HPV 18+宫颈癌HeLa细胞的抗增殖活性的机制仍然未知。本研究比较研究了Caerin 1.1、1.9及其混合物对HeLa细胞的抗增殖作用，然后通过共聚焦显微镜检查来评估细胞摄入的肽。采用串联质量标签标记蛋白质组学技术，揭示了在细胞和细胞生长环境中受肽处理显著调节的蛋白质，以阐明被调节的信号通路。进行蛋白质印迹以确认通路的调节。与未处理的和对照肽相比，蛙皮素1.1和1.9都高度抑制HeLa细胞增殖，具有显著的累加效应。它们以不同的幅度进入细胞。在显著上调的蛋白质中检测到强烈的蛋白质-蛋白质相互作用。蛋白质的翻译、折叠和定位以及RNA加工、凋亡过程在处理后显著富集。细胞凋亡信号传导被认为是肿瘤坏死因子-α（TNF-α）途径激活的结果，由caspase 3和caspase 9的剂量依赖性升高水平指示。表皮生长因子受体和雄激素受体途径似乎受到肽的抑制。此外，从定量结果得到的T细胞受体的活化进一步暗示了在处理后将更多的T细胞募集到细胞生长环境中的可能性，并且对T细胞介导的HeLa细胞杀伤更敏感。我们的研究结果表明，caerin 1.1和1.9介导的HeLa细胞的凋亡信号，并可能随后增强适应性T细胞免疫反应。
Host defense caerin 1.1 and 1.9 peptides, isolated from the glandular secretion of Australian tree frogs, the genusLitoria, have been previously shown to have multiple biological activities, including the inhibition of human papillomavirus (HPV) 16 early protein E7 transformed murine as well as human cancerous cell proliferation bothin vitroandin vivo. However, the mechanism underlying their anti-proliferative activities against HPV18+ cervical cancer HeLa cells remains unknown. This study comparatively investigated the anti-proliferation on HeLa cells by caerin 1.1, 1.9, and their mixture, followed by confocal microscopy examination to assess the cellular intake of the peptides. Tandem mass tag labeling proteomics was employed to reveal the proteins that were significantly regulated by the peptide treatment in cells and cell growth environment, to elucidate the signaling pathways that were modulated. Western blot was performed to confirm the modulation of the pathways. Both caerin 1.1 and 1.9 highly inhibited HeLa cell proliferation with a significant additive effect compared to untreated and control peptide. They entered the cells with different magnitudes. Intensive protein-protein interaction was detected among significantly upregulated proteins. Translation, folding and localization of proteins and RNA processing, apoptosis process was significantly enriched post the treatments. The apoptotic signaling was suggested as a result of tumor necrosis factor-alpha (TNF-alpha) pathway activation, indicated by the dose-dependent elevated levels of caspase 3 and caspase 9. The epidermal growth factor receptor and androgen receptor pathways appeared inhibited by the peptides. Moreover, the activation of T-cell receptor derived from the quantitation results further implies the likelihood of recruiting more T cells to the cell growth environment post the treatment and more sensitive to T cell mediated killing of HeLa cells. Our results indicate that caerin 1.1 and 1.9 mediate apoptotic signals of HeLa cells and may subsequently enhances adaptive T cell immune responses.