title: In vitro antiproliferative activity of 2,3-dihydroxy-9,10-anthraquinone induced apoptosis against COLO320 cells through cytochrome c release caspase mediated pathway with PI3K/AKT and COX-2 inhibition.
abstract: The present study investigated the anticancer activity of 2,3-dihydroxy-9,10-anthraquinone against different cancer cells such as MCF-7, COLO320, HepG-2, Skov-3, MOLM-14, NB-4, CEM, K562, Jurkat, HL-60, U937, IM-9 and Vero. 2,3-dihydroxy-9,10-anthraquinone showed good antiproliferative activity against COLO320 cells when compared to other tested cells. The cytotoxicity results showed 79.8% activity at the dose of 2.07 μM with IC50 value of 0.13 μM at 24 h in COLO320 cells. So we chose COLO320 cells for further anticancer studies. mRNA expression was confirmed by qPCR analysis using SYBR green method. Treatment with 2,3-dihydroxy-9,10-anthraquinone was found to trigger intrinsic apoptotic pathway as indicated by down regulation of Bcl-2, Bcl-xl; up regulation of Bim, Bax, Bad; release of cytochrome c and pro-caspases cleaving to caspases. Furthermore, 2,3-dihydroxy-9,10-anthraquinone stopped at G0/G1 phase with modulation in protein levels of cyclins. On the other hand PI3K/AKT signaling plays an important role in cell metabolism. We found that 2,3-dihydroxy-9,10-anthraquinone inhibits PI3K/AKT activity after treatment. Also, COX-2 enzyme plays a major role in colorectal cancer. Our results showed that the treatment significantly reduced COX-2 enzyme in COLO320 cells. These results indicated antiproliferative activity of 2,3-dihydroxy-9,10-anthraquinone involving apoptotic pathways, mitochondrial functions, cell cycle checkpoint and controlling the over expression genes during the colorectal cancer. Molecular docking studies showed that the compound bound stably to the active sites of Bcl-2, COX-2, PI3K and AKT. This is the first report of anticancer mechanism involving 2,3-dihydroxy-9,10-anthraquinone in COLO320 cells. The present results might provide helpful suggestions for the design of antitumor drugs toward colorectal cancer treatment.
authors.affiliation: Department of Hematology, Fujita Health University, 1-98, Dengakugakubo, Kutsukake-cho, Toyoake, Aichi, 470-1192, Japan; Division of Cancer Biology, Entomology Research Institute, Loyola College, Chennai, 600 034, India. Electronic address: balasang@fujita-hu.ac.jp.
authors.forename: C
authors.initials: C
authors.lastname: Balachandran
authors.affiliation: Department of Hematology, Fujita Health University, 1-98, Dengakugakubo, Kutsukake-cho, Toyoake, Aichi, 470-1192, Japan.
authors.forename: N
authors.initials: N
authors.lastname: Emi
authors.affiliation: Organic & Bio-organic Chemistry Laboratory, CSIR-Central Leather Research Institute, Chennai, 600 020, India.
authors.forename: Y
authors.initials: Y
authors.lastname: Arun
authors.affiliation: Laboratory of Molecular Biology, Institute of Joint Research​, Fujita Health University, 1-98, Dengakugakubo, Kutsukake-cho, Toyoake, Aichi, 470-1192, Japan.
authors.forename: N
authors.initials: N
authors.lastname: Yamamoto
authors.affiliation: Division of Cancer Biology, Entomology Research Institute, Loyola College, Chennai, 600 034, India; Department of Botany and Microbiology, Addiriya Chair for Environmental Studies, College of Science, King Saud University, P.O.Box.2455, Riyadh, 11451, Saudi Arabia.
authors.forename: V
authors.initials: V
authors.lastname: Duraipandiyan
authors.affiliation: Department of Hematology, Fujita Health University, 1-98, Dengakugakubo, Kutsukake-cho, Toyoake, Aichi, 470-1192, Japan.
authors.forename: Yoko
authors.initials: Y
authors.lastname: Inaguma
authors.affiliation: Department of Hematology, Fujita Health University, 1-98, Dengakugakubo, Kutsukake-cho, Toyoake, Aichi, 470-1192, Japan.
authors.forename: Akinao
authors.initials: A
authors.lastname: Okamoto
authors.affiliation: Division of Cancer Biology, Entomology Research Institute, Loyola College, Chennai, 600 034, India; Visiting Professor Program, Deanship of Scientific Research, College of Science, King Saud Univeristy, Saudi Arabia.
authors.forename: S
authors.initials: S
authors.lastname: Ignacimuthu
authors.affiliation: Department of Botany and Microbiology, Addiriya Chair for Environmental Studies, College of Science, King Saud University, P.O.Box.2455, Riyadh, 11451, Saudi Arabia.
authors.forename: N A
authors.initials: NA
authors.lastname: Al-Dhabi
authors.affiliation: Organic & Bio-organic Chemistry Laboratory, CSIR-Central Leather Research Institute, Chennai, 600 020, India.
authors.forename: P T
authors.initials: PT
authors.lastname: Perumal
mesh_terms: D000880:Anthraquinones / Q000494:pharmacology*; D000970:Antineoplastic Agents / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects*; D020169:Caspases / Q000378:metabolism*; D059447:Cell Cycle Checkpoints / Q000187:drug effects; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects*; D051546:Cyclooxygenase 2 / Q000378:metabolism*; D052246:Cyclooxygenase 2 Inhibitors / Q000494:pharmacology; D045304:Cytochromes c / Q000378:metabolism*; D016193:G1 Phase / Q000187:drug effects; D018922:HL-60 Cells; D056945:Hep G2 Cells; D006801:Humans; D020014:K562 Cells; D061986:MCF-7 Cells; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism; D062105:Molecular Docking Simulation / Q000379:methods; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism*; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism*; D016192:Resting Phase, Cell Cycle / Q000187:drug effects; D015398:Signal Transduction / Q000187:drug effects; D020298:U937 Cells
chemical_list: D000880:Anthraquinones; D000970:Antineoplastic Agents; D052246:Cyclooxygenase 2 Inhibitors; C042834:9,10-anthraquinone; D045304:Cytochromes c; D051546:Cyclooxygenase 2; C496540:PTGS2 protein, human; D019869:Phosphatidylinositol 3-Kinases; D051057:Proto-Oncogene Proteins c-akt; D020169:Caspases
keywords: 2,3-dihydroxy-9,10-anthraquinone; AKT/PI3K; COX-2; Caspases; Molecular docking