PMID- 26339797 OWN - NLM STAT- MEDLINE DCOM- 20160517 LR - 20181113 IS - 1932-6203 (Electronic) IS - 1932-6203 (Linking) VI - 10 IP - 9 DP - 2015 TI - The Apoptotic Effect of HIF-1alpha Inhibition Combined with Glucose plus Insulin Treatment on Gastric Cancer under Hypoxic Conditions. PG - e0137257 LID - 10.1371/journal.pone.0137257 [doi] LID - e0137257 AB - Gastric cancer grows under a hypoxic environment. HIF-1alpha is known to play an important role in controlling the production of reactive oxygen species (ROS) in the mitochondria under hypoxic conditions. We previously established HIF-1alpha knockdown (KD) cells and control (SC) cells in the 58As9 gastric cancer cell line. In this study, we revealed that KD cells, but not SC cells, induced apoptosis under conditions of hypoxia (1% O2) due to excessive production of ROS. A quantitative RT-PCR analysis demonstrated that the expressions of ten genes, which are involved in the control mechanisms of ROS (including the Warburg effect, mitophagy, electron transport chain [ETC] modification and ROS scavenging), were regulated by HIF-1alpha. Moreover, the promotion of glucose uptake by glucose plus insulin (GI) treatment enhanced the apoptotic effect, which was accompanied by further ROS production in hypoxic KD cells. A Western blot analysis showed that the membranous expression of GLUT1 in KD cells was elevated by glucose and/or insulin treatments, indicating that the GI-induced glucose uptake is mediated by the increased translocation of GLUT1 on the cell membrane. Finally, the anti-tumor effect of HIF-1alpha knockdown (KD) plus GI was evaluated using a tumor xenograft model, where a hypoxic environment naturally exists. As a result, the GI treatment strongly inhibited the growth of the KD tumors whereby cell apoptosis was highly induced in comparison to the control treatment. In contrast, the growth of the SC tumors expressing HIF-1alpha was not affected by the GI treatment. Taken together, the results suggest that HIF-1alpha inhibition plus GI may be an ideal therapy, because the apoptosis due to the destruction of ROS homeostasis is specifically induced in gastric cancer that grows under a hypoxic environment, but not in the normal tissue under the aerobic conditions. FAU - Tanaka, Tomokazu AU - Tanaka T AD - Department of Surgery, Saga University Faculty of Medicine, Nabeshima, Saga, Japan. FAU - Kitajima, Yoshihiko AU - Kitajima Y AD - Department of Surgery, Saga University Faculty of Medicine, Nabeshima, Saga, Japan; Department of Surgery, NHO Higashisaga Hospital, Harakoga, Miyaki-town, Miyaki-Gun, Saga, Japan. FAU - Miyake, Shuusuke AU - Miyake S AD - Department of Surgery, Saga University Faculty of Medicine, Nabeshima, Saga, Japan. FAU - Yanagihara, Kazuyoshi AU - Yanagihara K AD - Division of Pathology, Research Center of Innovative Oncology, National Cancer Center Hospital East, Kashiwanoha, Kashiwa, Chiba, Japan. FAU - Hara, Hiromitsu AU - Hara H AD - Department of Immunology, Kagoshima University Graduate School of Medical and Dental Sciences, Sakuragaoka, Kagoshima, Japan. FAU - Nishijima-Matsunobu, Aki AU - Nishijima-Matsunobu A AD - Department of Molecular and Tumor Pathology, Akita University Graduate School of Medicine, Hondo, Akita, Japan. FAU - Baba, Koichi AU - Baba K AD - Department of Surgery, Saga University Faculty of Medicine, Nabeshima, Saga, Japan. FAU - Shida, Masaaki AU - Shida M AD - Department of Surgery, Saga University Faculty of Medicine, Nabeshima, Saga, Japan. FAU - Wakiyama, Kota AU - Wakiyama K AD - Department of Surgery, Saga University Faculty of Medicine, Nabeshima, Saga, Japan. FAU - Nakamura, Jun AU - Nakamura J AD - Department of Surgery, Saga University Faculty of Medicine, Nabeshima, Saga, Japan. FAU - Noshiro, Hirokazu AU - Noshiro H AD - Department of Surgery, Saga University Faculty of Medicine, Nabeshima, Saga, Japan. LA - eng PT - Journal Article DEP - 20150904 PL - United States TA - PLoS One JT - PloS one JID - 101285081 RN - 0 (Glucose Transporter Type 1) RN - 0 (HIF1A protein, human) RN - 0 (Hypoxia-Inducible Factor 1, alpha Subunit) RN - 0 (Insulin) RN - 0 (Neoplasm Proteins) RN - 0 (RNA, Small Interfering) RN - 0 (Reactive Oxygen Species) RN - 0 (SLC2A1 protein, human) RN - 0 (VEGFA protein, human) RN - 0 (Vascular Endothelial Growth Factor A) RN - IY9XDZ35W2 (Glucose) RN - S88TT14065 (Oxygen) SB - IM MH - Animals MH - Apoptosis/drug effects MH - Cell Hypoxia MH - Cell Line, Tumor MH - Female MH - *Gene Expression Regulation, Neoplastic MH - Gene Knockdown Techniques MH - Glucose/*pharmacology MH - Glucose Transporter Type 1/genetics/metabolism MH - Humans MH - Hypoxia/*drug therapy/genetics/metabolism/pathology MH - Hypoxia-Inducible Factor 1, alpha Subunit/*antagonists & inhibitors/genetics/metabolism MH - Insulin/*pharmacology MH - Mice MH - Mice, Nude MH - Neoplasm Proteins/genetics/metabolism MH - Oxygen/pharmacology MH - RNA, Small Interfering/genetics/metabolism MH - Reactive Oxygen Species/agonists/metabolism MH - Signal Transduction MH - Stomach Neoplasms/*drug therapy/genetics/metabolism/pathology MH - Vascular Endothelial Growth Factor A/genetics/metabolism MH - Xenograft Model Antitumor Assays PMC - PMC4560388 COIS- Competing Interests: The authors have declared that no competing interests exist. EDAT- 2015/09/05 06:00 MHDA- 2016/05/18 06:00 PMCR- 2015/09/04 CRDT- 2015/09/05 06:00 PHST- 2015/04/30 00:00 [received] PHST- 2015/08/13 00:00 [accepted] PHST- 2015/09/05 06:00 [entrez] PHST- 2015/09/05 06:00 [pubmed] PHST- 2016/05/18 06:00 [medline] PHST- 2015/09/04 00:00 [pmc-release] AID - PONE-D-15-16624 [pii] AID - 10.1371/journal.pone.0137257 [doi] PST - epublish SO - PLoS One. 2015 Sep 4;10(9):e0137257. doi: 10.1371/journal.pone.0137257. eCollection 2015.