title: Effects and mechanisms of anti-CD44 monoclonal antibody A3D8 on proliferation and apoptosis of sphere-forming cells with stemness from human ovarian cancer.
abstract: OBJECTIVE
  CD44(+) human ovarian cancer stem cells (CSCs) and CSC-like cells have been identified and characterized. Compelling evidence has revealed that CD44 is involved in the occurrence and development of cancers. Our previous study showed that sphere-forming cells (SFCs) from the human ovarian cancer cell line SKOV-3 had CSC capacity. Therefore, in the present study, we aimed to investigate the effects and mechanisms of the anti-CD44 monoclonal antibody A3D8 on the proliferation and apoptosis of SFCs to explore novel strategies for the treatment of ovarian cancer.
  
  
  METHODS
  We investigated the effects and mechanisms of A3D8 on the proliferation and apoptosis of SFCs using the MTS assay, cell cycle analysis, an annexin V-fluorescein isothiocyanate/propidium iodide kit, Rh123 apoptosis detection kit, real-time reverse transcription polymerase chain reaction and Western blotting.
  
  
  RESULTS
  After CD44 ligation by A3D8, SFC cell proliferation was notably attenuated, cell cycle progression was arrested in the S phase, and apoptosis was significantly increased. The effect of A3D8 was enhanced in a dose- and time-dependent manner, and the effect of apoptosis induction by DDP was enhanced by combination treatment with A3D8. Furthermore, the messenger RNA expression levels of p21 and caspase-3 were up-regulated, whereas those of CDK2, cyclinA, and Bcl-2 were down-regulated. The protein expression levels of caspase-3 were up-regulated, whereas those of CDK2, cyclinA, and Bcl-2 were down-regulated.
  
  
  CONCLUSIONS
  Our findings indicate that anti-CD44 monoclonal antibodies may be a potential strategy for the treatment of human ovarian cancer after conventional therapy via inhibition of growth and the promotion of apoptosis in SFCs with stemness.
authors.affiliation: *Department of Immunology of Tianjin Medical University and Tianjin Key Laboratory of Cellular and Molecular Immunology and Key Laboratory of Immune Microenvironment and Disease of Ministry of Education; †Reproductive Medicine Centre, Tianjin Central Hospital for Obstetrics & Gynecology; and ‡Key laboratory of Hormone and Development, Ministry of Health of China and Metabolic Disease Hospital/Institute of Endocrinology, Tianjin Medical University, Tianjin, China.
authors.forename: Yong-Rui
authors.initials: YR
authors.lastname: Du
authors.forename: Ying
authors.initials: Y
authors.lastname: Chen
authors.forename: Yan
authors.initials: Y
authors.lastname: Gao
authors.forename: Xiu-Long
authors.initials: XL
authors.lastname: Niu
authors.forename: Ya-Jing
authors.initials: YJ
authors.lastname: Li
authors.forename: Wei-Min
authors.initials: WM
authors.lastname: Deng
mesh_terms: D000230:Adenocarcinoma / Q000188:drug therapy* / Q000473:pathology; D000911:Antibodies, Monoclonal / Q000494:pharmacology / Q000627:therapeutic use*; D017209:Apoptosis / Q000187:drug effects; D001201:Ascites / Q000473:pathology; D053148:Caspase 3 / Q000378:metabolism; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D002945:Cisplatin; D019925:Cyclin A / Q000378:metabolism; D051357:Cyclin-Dependent Kinase 2 / Q000378:metabolism; D050759:Cyclin-Dependent Kinase Inhibitor p21 / Q000378:metabolism; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D018960:Hyaluronan Receptors / Q000276:immunology; D014411:Neoplastic Stem Cells / Q000187:drug effects* / Q000378:metabolism; D010051:Ovarian Neoplasms / Q000188:drug therapy* / Q000473:pathology; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; C497483:CD44 protein, human; C493956:CDKN1A protein, human; D019925:Cyclin A; D050759:Cyclin-Dependent Kinase Inhibitor p21; D018960:Hyaluronan Receptors; D019253:Proto-Oncogene Proteins c-bcl-2; C495894:CDK2 protein, human; D051357:Cyclin-Dependent Kinase 2; D053148:Caspase 3; D002945:Cisplatin