title: Detection of a specific pattern of hyaluronan oligosaccharides and their binding proteins in human ovarian tumour.
abstract: Tumour cells generate hyaluronan (HA) oligomers (O-HA) by an autocrine mechanism to regulate their own behaviour through receptor interaction, necessitating analysis of HA sizes and its receptor expression in tumour progression. In this study for the first time, we identified specific size of HA in malignant ovarian tumour compared to benign tumour tissue. Therefore, we prepared the identified HA probes and conducted multiplex and monoplex ligand blot analysis and Immunohistochemistry to identify their receptor expression and distribution. Although, HA recognized CD44 as principle receptors despite of size, multiplex analysis showed multiple receptor expression with distribution at the tumour cell surface. Furthermore, the HA 6-mer (major O-HA of ovarian tumour) pull down of tumour tissue proteins showed 120 kDa protein along with CD44 with over expression in the malignant tumour. Upon depletion of CD44 protein HA 6-mer showed a major 120 kDa protein with distribution at nuclear membrane, suggesting that this protein may play an important role in ovarian tumour progression. In summary, ovarian tumour cells of different grade showed heterogeneity in generation of HA oligomers and their interaction with specific receptors. Therefore, simultaneous analysis of O-HA and their receptors expression could serve as a prognostic indicator during tumorigenesis. Copyright © 2016 John Wiley & Sons, Ltd.
  
  
  CONCLUSIONS
  Down regulation of hyaluronan (HA) at tumour epithelial cells results in the generation of O-HA. Increasing evidence about O-HA biological functions (mainly in vitro) are available, but we lack information of O-HA sizes generated during tumorigenesis and less information is available about their receptor interaction. We used biochemical approaches and identified that tumour cells of different grade possessing heterogeneity in generation of O-HA and specificity towards receptor binding. We identified a new receptor for HA 6-mer that over express in cancer tissue that shows its role in tumour progression. Collectively, simultaneous identification of HA sizes and their receptors could serve as a prognostic marker.
authors.affiliation: Department of Studies in Biochemistry, University of Mysore, Mysore, Karnataka, India.|Yenepoya Research Centre, Yenepoya University, Mangalore, Karnataka, India.
authors.forename: Srinivasa Prasad
authors.initials: SP
authors.lastname: Kolapalli
authors.affiliation: Department of Studies in Biochemistry, University of Mysore, Mysore, Karnataka, India.
authors.forename: Venatrao
authors.initials: V
authors.lastname: Nunna
authors.affiliation: Bharath Cancer Hospital, Mysore, Karnataka, India.
authors.forename: Anil
authors.initials: A
authors.lastname: Thomas
authors.affiliation: Department of Studies in Biochemistry, University of Mysore, Mysore, Karnataka, India.
authors.forename: Karuna Kumar
authors.initials: KK
authors.lastname: Mortha
authors.affiliation: Department of Anatomy and Cellular Biology, Tufts University - School of Medicine, Boston, MA, USA.
authors.forename: Shib Das
authors.initials: SD
authors.lastname: Banerjee
authors.affiliation: Rutgers Cancer Institute of New Jersey, Department of Medicine, Division of Medical Oncology - Rutgers, The State University of New Jersey, Robert Wood Johnson Medical School, New Brunswick, NJ, USA.
authors.forename: Rajeev K
authors.initials: RK
authors.lastname: Boregowda
mesh_terms: D002352:Carrier Proteins / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism; D002467:Cell Nucleus / Q000378:metabolism; D005260:Female; D006801:Humans; D018960:Hyaluronan Receptors / Q000378:metabolism; D006820:Hyaluronic Acid / Q000378:metabolism*; D007150:Immunohistochemistry; D008970:Molecular Weight; D009363:Neoplasm Proteins / Q000378:metabolism*; D009844:Oligosaccharides / Q000378:metabolism*; D010051:Ovarian Neoplasms / Q000378:metabolism*; D012015:Reference Standards
chemical_list: C497483:CD44 protein, human; D002352:Carrier Proteins; D018960:Hyaluronan Receptors; D009363:Neoplasm Proteins; D009844:Oligosaccharides; D006820:Hyaluronic Acid
keywords: CD44; HA 6-mers; HA oligosaccharides; hyaluronic acid; ovarian tumour