title: Amplification and overexpression of aurora kinase A (AURKA) in immortalized human ovarian epithelial (HOSE) cells.
abstract: Immortalization is an early and essential step of human carcinogenesis. Amplification of chromosome 20q has been shown to be a common event in immortalized cells and cancers. We have previously reported that gain and amplification of chromosome 20q is a non-random and common event in immortalized human ovarian surface epithelial (HOSE) cells. The chromosome 20q harbors genes including TGIF2 (20q11.2-q12), AIB1 (20q12), PTPN1 (20q13.1), ZNF217 (20q13.2), and AURKA (20q13.2-q13.3), which were previously reported to be amplified and overexpressed in ovarian cancers. Some of these genes may be involved in immortalization of HOSE cells and represent crucial premalignant changes in ovarian surface epithelium. Investigation of the involvement of these genes was examined in four pairs of pre-crisis (preimmortalized) and post-crisis (immortalized) HOSE cells. Overexpression of AURKA (Aurora kinase A), also known as BTAK and STK15, by both real time-quantitative polymerase chain reaction (RT-QPCR) and Western blotting was detected in all the four immortalized HOSE cells examined while overexpression of AIB1 and ZNF217 was observed in two of four immortalized HOSE cells examined. Overexpression of TGIF2 and PTPN1 was not significant in our immortalized HOSE cell systems. The degree of overexpression of AURKA was shown to be closely associated with the amplification of chromosome 20q in immortalized HOSE cells. Fluorescence in situ hybridization (FISH) with labeled P1 artificial clone (PAC) confirmed the amplification of the chromosomal region (20q13.2-13.3) where AURKA resides. DNA amplification of AURKA was also confirmed using semi-quantitative PCR. Our study showed that amplification and overexpression of AURKA is a common and significant event during immortalization of HOSE cells and may represent an important premalignant change in ovarian carcinogenesis.
authors.affiliation: Cancer Biology Laboratory, Department of Anatomy, Faculty of Medicine, University of Hong Kong, Pokfulam, Hong Kong SAR, China.
authors.forename: C M
authors.initials: CM
authors.lastname: Chung
authors.forename: C
authors.initials: C
authors.lastname: Man
authors.forename: Y
authors.initials: Y
authors.lastname: Jin
authors.forename: C
authors.initials: C
authors.lastname: Jin
authors.forename: X Y
authors.initials: XY
authors.lastname: Guan
authors.forename: Q
authors.initials: Q
authors.lastname: Wang
authors.forename: T S K
authors.initials: TS
authors.lastname: Wan
authors.forename: A L M
authors.initials: AL
authors.lastname: Cheung
authors.forename: S W
authors.initials: SW
authors.lastname: Tsao
mesh_terms: D064096:Aurora Kinase A; D064127:Aurora Kinases; D001483:Base Sequence; D018797:Cell Cycle Proteins / Q000235:genetics*; D002478:Cells, Cultured; D002874:Chromosome Mapping; D002890:Chromosomes, Human, Pair 20 / Q000235:genetics*; D017931:DNA Primers; D004847:Epithelial Cells / Q000201:enzymology*; D005260:Female; D005784:Gene Amplification; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D010053:Ovary / Q000166:cytology / Q000201:enzymology*; D016133:Polymerase Chain Reaction; D011494:Protein Kinases / Q000235:genetics*; D017346:Protein Serine-Threonine Kinases; D020133:Reverse Transcriptase Polymerase Chain Reaction; D029867:Xenopus Proteins / Q000235:genetics*
chemical_list: D018797:Cell Cycle Proteins; D017931:DNA Primers; D029867:Xenopus Proteins; D011494:Protein Kinases; C112134:AURKA protein, Xenopus; C576917:AURKA protein, human; D064096:Aurora Kinase A; D064127:Aurora Kinases; D017346:Protein Serine-Threonine Kinases