Institute of Biochemistry and Biotechnology, Chung Shan Medical University, Taichung 402, Taiwan.
Peptides. 2011 Feb;32(2):342-52. doi: 10.1016/j.peptides.2010.11.003. Epub 2010 Nov 18.
The inhibitory function of tilapia hepcidin (TH)1-5, an antimicrobial peptide, was not examined in previous studies. In this study, we synthesized the TH1-5 peptide and tested TH1-5's antitumor activity against several tumor cell lines. We show that TH1-5 inhibited the proliferation of tumor cells and reduced colony formation in a soft agar assay. Scanning electron microscopy and transmission electron microscopy showed that TH1-5 altered the membrane structure similar to the function of a lytic peptide. Acridine orange/ethidium bromide staining, a wound-healing assay, and a flow cytometric analysis showed that TH1-5 induced necrosis with high-concentration treatment and induced apoptosis with low-concentration treatment. Inflammation is known to be closely associated with the development of cancer. TH1-5 showing anti-inflammatory effects in a previous publication induced us to evaluate the anti-inflammatory effects in cancer cell lines through the expressions of immune-related genes after being treated with the TH1-5 peptide. However, real-time qualitative RT-PCR indicated that TH1-5 treatment induced downregulation of the expressions of interleukin (IL)-6, IL-8, IL-12, IL-15, interferon-γ, CTSG, caspase-7, and Bcl-2, and upregulation of IL-2 and CAPN5 in HeLa cells, and upregulation of IL-8 and CTSG in HT1080 cells. These results suggest that TH1-5 possibly induces an inflammatory response in HeLa cells, but not in HT1080 cells. Overall, these results indicate that TH1-5 possesses the potential to be a novel peptide for cancer therapy.
先前的研究并未检测罗非鱼抗菌肽（hepcidin，Hep）1-5 的抑制功能。本研究合成了 Hep1-5 肽并测试了其对几种肿瘤细胞系的抗肿瘤活性。结果表明，Hep1-5 抑制了肿瘤细胞的增殖并降低了软琼脂中的集落形成。扫描电子显微镜和透射电子显微镜显示，Hep1-5 改变了膜结构，类似于溶细胞肽的功能。吖啶橙/溴化乙锭染色、划痕愈合实验和流式细胞术分析表明，Hep1-5 在高浓度处理时诱导坏死，在低浓度处理时诱导凋亡。众所周知，炎症与癌症的发展密切相关。Hep1-5 在之前的研究中显示出抗炎作用，这促使我们通过用 Hep1-5 肽处理后免疫相关基因的表达来评估其在肿瘤细胞系中的抗炎作用。然而，实时定量 RT-PCR 表明，Hep1-5 处理诱导 HeLa 细胞中白细胞介素（IL）-6、IL-8、IL-12、IL-15、干扰素-γ、CTSG、caspase-7 和 Bcl-2 的表达下调，以及 IL-2 和 CAPN5 的上调，HT1080 细胞中则 IL-8 和 CTSG 的上调。这些结果表明，Hep1-5 可能在 HeLa 细胞中诱导炎症反应，但在 HT1080 细胞中不会。总之，这些结果表明，Hep1-5 具有成为癌症治疗新型肽的潜力。
