PDF(Pubmed)
Abstract：
(1) Background: GHaK is derived from the antimicrobial peptide temporin-GHa by substituting the amino acid H with K to enhance its bactericidal activity. The present research aims to broaden the pharmacological potential of GHaK by exploring its antineoplastic activity against human lung adenocarcinoma. (2) Methods: The cell viability, migration, invasion, apoptosis, and cell cycle of A549 and PC-9 cells were tested after GHaK treatment. miRNA sequencing, RT-PCR, Western blotting, and luciferase reporter gene assay were further performed to reveal the potential mechanism. (3) Results: GHaK significantly suppressed cell viability, migration, and invasion; induced apoptosis; and caused cell cycle arrest in the G2/M and S phase in PC-9 and A549 cells, respectively. The miRNA sequencing results show a total of 161 up-regulated and 115 down-regulated miRNAs. Furthermore, the study identified six up-regulated miRNAs (miR-4516, miR-4284, miR-204-5p, miR-12136, miR-4463, and miR-1296-3p) and their inhibitory effects on the expressions of target genes (Wnt 8B, FZD2, DVL3, and FOSL1) caused by miR-4516 directly interacting with Wnt 8B. Western blotting revealed the down-regulation of p-GSK-3β, along with a decreased expressions of cyclin A1 and CDK2 in A549 cells and cyclin B1 and CDK1 in PC-9 cells. (4) Conclusions: Temporin-GHaK exhibits antineoplastic activity against human lung adenocarcinoma by inhibiting the Wnt signaling pathway through miRNA-4516.
摘要：
（1）背景：GHaK是由抗菌肽temporin-Gha通过用K取代氨基酸H来增强其杀菌活性而衍生的。本研究旨在通过探索GHaK对人肺腺癌的抗肿瘤活性来拓宽其药理潜力。(2)方法:细胞活力,迁移,入侵,凋亡，GHaK处理后检测A549和PC-9细胞的细胞周期。miRNA测序，RT-PCR,西方印迹,和荧光素酶报告基因检测进一步揭示了潜在的机制。(3)结果:GHaK显著抑制细胞活力,迁移,和侵袭；诱导凋亡；并导致PC-9和A549细胞在G2/M和S期的细胞周期停滞，分别。miRNA测序结果显示总共161个上调的miRNA和115个下调的miRNA。此外，该研究确定了六个上调的miRNA(miR-4516，miR-4284，miR-204-5p，miR-12136、miR-4463和miR-1296-3p)及其对靶基因表达的抑制作用(Wnt8B,FZD2，DVL3和FOSL1）由miR-4516直接与Wnt8B相互作用引起。Western印迹显示p-GSK-3β的下调，A549细胞中细胞周期蛋白A1和CDK2的表达以及PC-9细胞中细胞周期蛋白B1和CDK1的表达降低。(4)结论:Temporin-GHaK通过miRNA-4516抑制Wnt信号通路对人肺腺癌具有抗肿瘤活性。
