title: KU-0060648 inhibits hepatocellular carcinoma cells through DNA-PKcs-dependent and DNA-PKcs-independent mechanisms.
abstract: Here we tested anti-tumor activity of KU-0060648 in preclinical hepatocellular carcinoma (HCC) models. Our results demonstrated that KU-0060648 was anti-proliferative and pro-apoptotic in established (HepG2, Huh-7 and KYN-2 lines) and primary human HCC cells, but was non-cytotoxic to non-cancerous HL-7702 hepatocytes. DNA-PKcs (DNA-activated protein kinase catalytic subunit) is an important but not exclusive target of KU-0060648. DNA-PKcs knockdown or dominant negative mutation inhibited HCC cell proliferation. On the other hand, overexpression of wild-type DNA-PKcs enhanced HepG2 cell proliferation. Importantly, KU-0060648 was still cytotoxic to DNA-PKcs-silenced or -mutated HepG2 cells, although its activity in these cells was relatively weak. Further studies showed that KU-0060648 inhibited PI3K-AKT-mTOR activation, independent of DNA-PKcs. Introduction of constitutively-active AKT1 (CA-AKT1) restored AKT-mTOR activation after KU-0060648 treatment in HepG2 cells, and alleviated subsequent cytotoxicity. In vivo, intraperitoneal (i.p.) injection of KU-0060648 significantly inhibited HepG2 xenograft growth in nude mice. AKT-mTOR activation was also inhibited in xenografted tumors. Finally, we showed that DNA-PKcs expression was significantly upregulated in human HCC tissues. Yet miRNA-101, an anti-DNA-PKcs miRNA, was downregulated. Over-expression of miR-101 in HepG2 cells inhibited DNA-PKcs expression and cell proliferation. Together, these results indicate that KU-0060648 inhibits HCC cells through DNA-PKcs-dependent and -independent mechanisms.
authors.affiliation: Department of Oncology, Kunshan First People's Hospital Affiliated to Jiangsu University, Kunshan 215300, China.
authors.forename: Min-Bin
authors.initials: MB
authors.lastname: Chen
authors.affiliation: Department of Orthopedics, the Second Affiliated Hospital of Soochow University, Suzhou 215000, China.
authors.forename: Zhen-Tao
authors.initials: ZT
authors.lastname: Zhou
authors.affiliation: Department of Breast Surgery, the Third Affiliated Hospital of Soochow University, Changzhou 213003, China.
authors.forename: Lan
authors.initials: L
authors.lastname: Yang
authors.affiliation: Department of Traditional Chinese Medicine, First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China.
authors.forename: Mu-Xin
authors.initials: MX
authors.lastname: Wei
authors.affiliation: Department of Oncology, Kunshan First People's Hospital Affiliated to Jiangsu University, Kunshan 215300, China.
authors.forename: Min
authors.initials: M
authors.lastname: Tang
authors.affiliation: Department of Medical Oncology, Wuxi People's Hospital Affiliated to Nanjing Medical University, Wuxi 214023, China.
authors.forename: Ting-Yan
authors.initials: TY
authors.lastname: Ruan
authors.affiliation: Department of Medical Oncology, Wuxi People's Hospital Affiliated to Nanjing Medical University, Wuxi 214023, China.
authors.forename: Jun-Ying
authors.initials: JY
authors.lastname: Xu
authors.affiliation: Department of Orthopedics, the Second Affiliated Hospital of Soochow University, Suzhou 215000, China.
authors.forename: Xiao-Zhong
authors.initials: XZ
authors.lastname: Zhou
authors.affiliation: Department of Neurosurgery, the First Affiliated Hospital of Soochow University, Suzhou 215006, China.
authors.forename: Gang
authors.initials: G
authors.lastname: Chen
authors.affiliation: Department of Medical Oncology, Wuxi People's Hospital Affiliated to Nanjing Medical University, Wuxi 214023, China.
authors.forename: Pei-Hua
authors.initials: PH
authors.lastname: Lu
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D006528:Carcinoma, Hepatocellular / Q000378:metabolism / Q000473:pathology*; D049109:Cell Proliferation / Q000187:drug effects; D002867:Chromones / Q000494:pharmacology*; D051747:DNA-Activated Protein Kinase / Q000378:metabolism; D056945:Hep G2 Cells; D006801:Humans; D008113:Liver Neoplasms / Q000378:metabolism / Q000473:pathology*; D051379:Mice; D008819:Mice, Nude; D035683:MicroRNAs / Q000378:metabolism; D009687:Nuclear Proteins / Q000378:metabolism; D013876:Thiophenes / Q000494:pharmacology*; D023041:Xenograft Model Antitumor Assays
chemical_list: C576809:2-(4-ethylpiperazin-1-yl)-N-(4-(2-morpholino-4-oxo-4H-chromen-8-yl)dibenzo(b,d)thiophen-1-yl)acetamide; D000970:Antineoplastic Agents; D002867:Chromones; C534072:MIRN101 microRNA, human; D035683:MicroRNAs; D009687:Nuclear Proteins; D013876:Thiophenes; D051747:DNA-Activated Protein Kinase; C497263:PRKDC protein, human
keywords: DNA-PKcs; KU-0060648; PI3K-AKT-mTOR signaling; hepatocellular carcinoma (HCC); miRNA