Phytochemical naphtho[1,2-b] furan-4,5‑dione induced topoisomerase II-mediated DNA damage response in human non-small-cell lung cancer

• Published in: Phytomedicine (Stuttgart) 2019-02, Vol.54, p.109-119
• Main Authors: Chien, Ching-Ming, Yang, Juan-Cheng, Wu, Pin-Hsuan, Wu, Chang-Yi, Chen, Guan-Yu, Wu, Yang-Chang, Chou, Chon-Kit, Tseng, Chih-Hua, Chen, Yeh-Long, Wang, Li-Fang, Chiu, Chien-Chih
• Format: Article
• Language: English
• Subjects: Animals; Antineoplastic Agents, Phytogenic - chemistry; Antineoplastic Agents, Phytogenic - pharmacology; Apoptosis; Apoptosis - drug effects; Carcinoma, Non-Small-Cell Lung - drug therapy; Carcinoma, Non-Small-Cell Lung - genetics; Cell Line, Tumor; DNA damage; DNA Damage - drug effects; DNA Topoisomerases, Type II - chemistry; DNA Topoisomerases, Type II - metabolism; Female; Furans - chemistry; Furans - pharmacology; Humans; Lung Neoplasms - drug therapy; Lung Neoplasms - genetics; Mice, Nude; Molecular Docking Simulation; Naphthoquinone; Naphthoquinones - chemistry; Naphthoquinones - pharmacology; NF-kappa B - metabolism; NF-Κb; Non-small cell lung cancer; Poly-ADP-Ribose Binding Proteins - antagonists & inhibitors; Poly-ADP-Ribose Binding Proteins - chemistry; Poly-ADP-Ribose Binding Proteins - metabolism; Reactive Oxygen Species - metabolism; Signal Transduction - drug effects; Topoisomerase II activity; Xenograft Model Antitumor Assays
• ISSN: 0944-7113; 1618-095X
• DOI: 10.1016/j.phymed.2018.06.025

Phytochemical naphtho[1,2-b] furan-4,5‑dione (NFD) presenting in Avicennia marina exert anti-cancer effects, but little is known regarding about DNA damage-mediated apoptosis in non-small-cell lung carcinoma (NSCLC). To examine whether NFD-induced apoptosis of NSCLC cells is correlated with the induction of DNA damage, and to investigate its underlying mechanism. The anti-proliferative effects of NFD were assessed by MTS Assay Kit FACS assay, and in vivo nude mice xenograft assay. The DNA damage related proteins, the Bcl-2 family and pro-apoptotic factors were examined by immunofluorescence assay, q-PCR, and western blotting. The activity of NF-κB p65 in nuclear extracts was detected using a colorimetric DNA-binding ELISA assay. The inhibitory activity of topoisomerase II (TOPO II) was evaluated by molecular docking and TOPO II catalytic assay. NFD exerted selective cytotoxicity against NSCLC H1299, H1437 and A549 cells rather than normal lung-embryonated cells MRC-5. Remarkably, we found that NFD activated the hull marker and modulator of DNA damage repairs such as γ-H2AX, ATM, ATR, CHK1, and CHK2 probably caused by the accumulation of intracellular reactive oxygen species (ROS) and inhibition of TOPO II activity. Furthermore, the suppression of transcription factor NF-κB by NFD resulted in significantly decreased levels of pro-survival proteins including Bcl-2 family Bcl-2, Bcl-xL and Mcl-1 and the endogenous inhibitors of apoptosis XIAP and survivin in H1299 cells. Moreover, the nude mice xenograft assay further validated the suppression of H1299 growth by NFD, which is the first report for evaluating the anti-cancer effect of NFD in vivo. These findings provide a novel mechanism indicating the inhibition of TOPO II activity and NF-κB signaling by NFD, leading to DNA damage and apoptosis of NSCLC tumor cells. [Display omitted]