title: Midazolam induces cellular apoptosis in human cancer cells and inhibits tumor growth in xenograft mice.
abstract: Midazolam is a widely used anesthetic of the benzodiazepine class that has shown cytotoxicity and apoptosisinducing activity in neuronal cells and lymphocytes. This study aims to evaluate the effect of midazolam on growth of K562 human leukemia cells and HT29 colon cancer cells. The in vivo effect of midazolam was investigated in BALB/c-nu mice bearing K562 and HT29 cells human tumor xenografts. The results show that midazolam decreased the viability of K562 and HT29 cells by inducing apoptosis and S phase cell-cycle arrest in a concentration-dependent manner. Midazolam activated caspase-9, capspase-3 and PARP indicating induction of the mitochondrial intrinsic pathway of apoptosis. Midazolam lowered mitochondrial membrane potential and increased apoptotic DNA fragmentation. Midazolam showed reactive oxygen species (ROS) scavenging activity through inhibition of NADPH oxidase 2 (Nox2) enzyme activity in K562 cells. Midazolam caused inhibition of pERK1/2 signaling which led to inhibition of the anti-apoptotic proteins Bcl-XL and XIAP and phosphorylation activation of the pro-apoptotic protein Bid. Midazolam inhibited growth of HT29 tumors in xenograft mice. Collectively our results demonstrate that midazolam caused growth inhibition of cancer cells via activation of the mitochondrial intrinsic pathway of apoptosis and inhibited HT29 tumor growth in xenograft mice. The mechanism underlying these effects of midazolam might be suppression of ROS production leading to modulation of apoptosis and growth regulatory proteins. These findings present possible clinical implications of midazolam as an anesthetic to relieve pain during in vivo anticancer drug delivery and to enhance anticancer efficacy through its ROS-scavenging and pro-apoptotic properties.
authors.affiliation: Gachon Institute of Pharmaceutical Sciences, Gachon University, Incheon, 406-840, Korea.
authors.forename: Siddhartha Kumar
authors.initials: SK
authors.lastname: Mishra
authors.forename: Ju-Hee
authors.initials: JH
authors.lastname: Kang
authors.forename: Chang Woo
authors.initials: CW
authors.lastname: Lee
authors.forename: Seung Hyun
authors.initials: SH
authors.lastname: Oh
authors.forename: Jun Sun
authors.initials: JS
authors.lastname: Ryu
authors.forename: Yun Soo
authors.initials: YS
authors.lastname: Bae
authors.forename: Hwan Mook
authors.initials: HM
authors.lastname: Kim
mesh_terms: D000759:Adjuvants, Anesthesia / Q000494:pharmacology; D000818:Animals; D000970:Antineoplastic Agents / Q000378:metabolism / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects* / Q000235:genetics; D053148:Caspase 3 / Q000378:metabolism; D053453:Caspase 9 / Q000378:metabolism; D049109:Cell Proliferation / Q000187:drug effects; D005260:Female; D019073:HT29 Cells; D006801:Humans; D020014:K562 Cells; D008562:Membrane Glycoproteins / Q000378:metabolism; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects*; D051379:Mice; D008807:Mice, Inbred BALB C; D008874:Midazolam / Q000378:metabolism / Q000494:pharmacology*; D000074662:NADPH Oxidase 2; D019255:NADPH Oxidases / Q000378:metabolism; D009369:Neoplasms / Q000378:metabolism / Q000473:pathology*; D017382:Reactive Oxygen Species / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects* / Q000235:genetics; D023041:Xenograft Model Antitumor Assays
chemical_list: D000759:Adjuvants, Anesthesia; D000970:Antineoplastic Agents; D008562:Membrane Glycoproteins; D017382:Reactive Oxygen Species; C056558:CYBB protein, human; D000074662:NADPH Oxidase 2; D019255:NADPH Oxidases; D053148:Caspase 3; D053453:Caspase 9; D008874:Midazolam