PMID- 28341244 OWN - NLM STAT- MEDLINE DCOM- 20180105 LR - 20211204 IS - 1872-7573 (Electronic) IS - 0378-8741 (Linking) VI - 203 DP - 2017 May 5 TI - Imperatorin suppresses proliferation and angiogenesis of human colon cancer cell by targeting HIF-1alpha via the mTOR/p70S6K/4E-BP1 and MAPK pathways. PG - 27-38 LID - S0378-8741(16)32149-3 [pii] LID - 10.1016/j.jep.2017.03.033 [doi] AB - ETHNOPHARMACOLOGICAL RELEVANCE: Angelica dahurica is a commonly used traditional Chinese medicine to treat migraine headache, toothache and cancer. Imperatorin is an active natural furocoumarin component originating from Angelica dahurica and has been shown to exhibit multiple bioeffector functions, including anti-cancer activity. However, the mechanism by which imperatorin inhibits tumor growth is not fully understood. AIM OF THE STUDY: The aim of this study was to investigate the effectiveness of imperatorin as a treatment of cancer and to identify the underlying mechanisms of its anticancer activity. MATERIALS AND METHODS: HCT116, HeLa, and Hep3B cells were used in this study. Major assays were promoter-reporter gene assay, MTT, western blot analysis, immunofluorescence assay, reverse transcription-PCR (RT-PCR), flow cytometric analysis, clonogenic assay, EdU labeling and immunofluorescence, xenografted assay, and VEGF ELISA. RESULTS: We here demonstrated the effect of imperatorin on hypoxia-inducible factor-1 (HIF-1) activation. Imperatorin showed a potent inhibitory activity against HIF-1 activation induced by hypoxia in various human cancer cell lines. This compound markedly decreased the hypoxia-induced accumulation of HIF-1alpha protein dose-dependently, whereas it did not affect the expressions of HIF-1beta and topoisomerase-I (Topo-I). Further analysis revealed that imperatorin inhibited HIF-1alpha protein synthesis, without affecting the expression level of HIF-1alpha mRNA or degradation of HIF-1alpha protein. Moreover, the phosphorylation levels of mammalian target of rapamycin (mTOR), ribosomal protein S6 kinase (p70S6K), eIF4E binding protein-1 (4E-BP1), eukaryotic initiation factor 4E (eIF4E), extracellular signal-regulated kinase-1/2 (ERK1/2), SAPK/JNK and p38 were significantly suppressed by imperatorin. Furthermore, imperatorin prevented hypoxia-induced expression of HIF-1 target genes and flow cytometric analysis indicated that imperatorin induced G1 phase arrest in human colon cancer cell (HCT116). We found that imperatorin administration inhibits tumor growth and blocks tumor angiogenesis in a xenograft tumor model. CONCLUSIONS: These results show that imperatorin inhibited HIF-1alpha protein synthesis by downregulating the mTOR/p70S6K/4E-BP1 and MAPK pathways. These conclusions suggest that imperatorin is an effective inhibitor of HIF-1 and provide new perspectives into the mechanism of its anticancer activity. CI - Copyright (c) 2017 Elsevier Ireland Ltd. All rights reserved. FAU - Mi, Chunliu AU - Mi C AD - Key Laboratory of Natural Resources of Changbai Mountain & Functional Molecules, Ministry of education, Molecular Medicine Research Center, College of Pharmacy, Yanbian University, Yanji 133002, Jilin Province, China. FAU - Ma, Juan AU - Ma J AD - Key Laboratory of Natural Resources of Changbai Mountain & Functional Molecules, Ministry of education, Molecular Medicine Research Center, College of Pharmacy, Yanbian University, Yanji 133002, Jilin Province, China. FAU - Wang, Ke Si AU - Wang KS AD - Key Laboratory of Natural Resources of Changbai Mountain & Functional Molecules, Ministry of education, Molecular Medicine Research Center, College of Pharmacy, Yanbian University, Yanji 133002, Jilin Province, China. FAU - Zuo, Hong Xiang AU - Zuo HX AD - Key Laboratory of Natural Resources of Changbai Mountain & Functional Molecules, Ministry of education, Molecular Medicine Research Center, College of Pharmacy, Yanbian University, Yanji 133002, Jilin Province, China. FAU - Wang, Zhe AU - Wang Z AD - Key Laboratory of Natural Resources of Changbai Mountain & Functional Molecules, Ministry of education, Molecular Medicine Research Center, College of Pharmacy, Yanbian University, Yanji 133002, Jilin Province, China. FAU - Li, Ming Yue AU - Li MY AD - Key Laboratory of Natural Resources of Changbai Mountain & Functional Molecules, Ministry of education, Molecular Medicine Research Center, College of Pharmacy, Yanbian University, Yanji 133002, Jilin Province, China. FAU - Piao, Lian Xun AU - Piao LX AD - Key Laboratory of Natural Resources of Changbai Mountain & Functional Molecules, Ministry of education, Molecular Medicine Research Center, College of Pharmacy, Yanbian University, Yanji 133002, Jilin Province, China. FAU - Xu, Guang Hua AU - Xu GH AD - Key Laboratory of Natural Resources of Changbai Mountain & Functional Molecules, Ministry of education, Molecular Medicine Research Center, College of Pharmacy, Yanbian University, Yanji 133002, Jilin Province, China. FAU - Li, Xuezheng AU - Li X AD - Key Laboratory of Natural Resources of Changbai Mountain & Functional Molecules, Ministry of education, Molecular Medicine Research Center, College of Pharmacy, Yanbian University, Yanji 133002, Jilin Province, China. FAU - Quan, Zhe Shan AU - Quan ZS AD - Key Laboratory of Natural Resources of Changbai Mountain & Functional Molecules, Ministry of education, Molecular Medicine Research Center, College of Pharmacy, Yanbian University, Yanji 133002, Jilin Province, China. Electronic address: zsquan@ybu.edu.cn. FAU - Jin, Xuejun AU - Jin X AD - Key Laboratory of Natural Resources of Changbai Mountain & Functional Molecules, Ministry of education, Molecular Medicine Research Center, College of Pharmacy, Yanbian University, Yanji 133002, Jilin Province, China. Electronic address: xjjin@ybu.edu.cn. LA - eng PT - Journal Article DEP - 20170321 PL - Ireland TA - J Ethnopharmacol JT - Journal of ethnopharmacology JID - 7903310 RN - 0 (Adaptor Proteins, Signal Transducing) RN - 0 (Angiogenesis Inhibitors) RN - 0 (Antineoplastic Agents, Phytogenic) RN - 0 (Cell Cycle Proteins) RN - 0 (EIF4EBP1 protein, human) RN - 0 (Furocoumarins) RN - 0 (Hypoxia-Inducible Factor 1, alpha Subunit) RN - 0 (Phosphoproteins) RN - EC 2.7.1.1 (MTOR protein, human) RN - EC 2.7.11.1 (Ribosomal Protein S6 Kinases, 70-kDa) RN - EC 2.7.11.1 (TOR Serine-Threonine Kinases) RN - K713N25C78 (imperatorin) SB - IM MH - Adaptor Proteins, Signal Transducing/metabolism MH - Angelica/*chemistry MH - Angiogenesis Inhibitors/isolation & purification/pharmacology MH - Animals MH - Antineoplastic Agents, Phytogenic/isolation & purification/*pharmacology MH - Cell Cycle Proteins MH - Cell Proliferation/drug effects MH - Colonic Neoplasms/blood supply/*drug therapy/pathology MH - Enzyme-Linked Immunosorbent Assay MH - Flow Cytometry MH - Furocoumarins/isolation & purification/*pharmacology MH - HCT116 Cells MH - HeLa Cells MH - Humans MH - Hypoxia-Inducible Factor 1, alpha Subunit/metabolism MH - MAP Kinase Signaling System/drug effects MH - Male MH - Mice MH - Mice, Inbred BALB C MH - Mice, Nude MH - Neovascularization, Pathologic/drug therapy/pathology MH - Phosphoproteins/metabolism MH - Ribosomal Protein S6 Kinases, 70-kDa/metabolism MH - TOR Serine-Threonine Kinases/metabolism MH - Xenograft Model Antitumor Assays OTO - NOTNLM OT - Antitumor OT - HIF-1alpha OT - Imperatorin OT - Translation EDAT- 2017/03/28 06:00 MHDA- 2018/01/06 06:00 CRDT- 2017/03/26 06:00 PHST- 2016/11/29 00:00 [received] PHST- 2017/03/17 00:00 [revised] PHST- 2017/03/20 00:00 [accepted] PHST- 2017/03/28 06:00 [pubmed] PHST- 2018/01/06 06:00 [medline] PHST- 2017/03/26 06:00 [entrez] AID - S0378-8741(16)32149-3 [pii] AID - 10.1016/j.jep.2017.03.033 [doi] PST - ppublish SO - J Ethnopharmacol. 2017 May 5;203:27-38. doi: 10.1016/j.jep.2017.03.033. Epub 2017 Mar 21.