title: Evaluating L1CAM expression in human endometrial cancer using qRT-PCR.
abstract: BACKGROUND
  Management of endometrial carcinoma (EC) still needs improvement of risk assessment. Recently, L1CAM immunohistochemical (IHC) evaluation showed a unique value to predict the outcome of early EC. However IHC results are often conflicting for lack of inter-laboratory standardisation.
  
  
  METHODS
  Here, as a proof of concept and to increase reproducibility we assayed eighty-two EC and 26 normal endometrium samples for L1CAM expression (L1CAMEXP) via qRT-PCR. The IHC evaluation was performed in 50 cancer samples. Moreover, we aimed to substantiate the in-vitro findings of L1CAM regulation through its promoter methylation (L1CAMMET), miR-34a expression and miR-34a promoter methylation. DNA methylation was assessed with MethyLight PCR technique.
  
  
  RESULTS
  High overall concordant results between IHC and RT-PCR evaluations were found. L1CAMEXP was detected in 11% of cancer specimens. These positive cancers exhibited a worse DFS (p=0.032) and OS (p=0.016) in a multivariate COX-regression model. L1CAMEXP predicted distant failure (p=0.007) and L1CAMMET predicted risk-reduction of lymph-node involvement (p=0.005). Inverse correlations between L1CAMEXP and L1CAMMET (p=0.004) and between L1CAMEXP and miR-34a expression (p=0.002) were found.
  
  
  CONCLUSIONS
  In conclusion qRT-PCR analysis is a reliable approach to evaluate L1CAM status in EC and L1CAMEXP was highly predictive for distant failure and poor outcome, confirming the large IHC-based studies. Interestingly, L1CAMMET was able to assess the risk of pelvic lymph-node involvement. Especially the latter finding has to be confirmed in larger prospective series.
authors.affiliation: Department of Gynecology and Obstetrics, Medical University of Innsbruck, Innsbruck, Austria.|Department of Gynecology and Obstetrics, University of Brescia, Brescia, Italy.
authors.forename: Sara
authors.initials: S
authors.lastname: Notaro
authors.affiliation: Department of Gynecology and Obstetrics, Medical University of Innsbruck, Innsbruck, Austria.
authors.forename: Daniel
authors.initials: D
authors.lastname: Reimer
authors.affiliation: Department of Gynecology and Obstetrics, Medical University of Innsbruck, Innsbruck, Austria.
authors.forename: Michaela
authors.initials: M
authors.lastname: Duggan-Peer
authors.affiliation: Department of Gynecology and Obstetrics, Medical University of Innsbruck, Innsbruck, Austria.
authors.forename: Heidi
authors.initials: H
authors.lastname: Fiegl
authors.affiliation: Department of Gynecology and Obstetrics, Medical University of Innsbruck, Innsbruck, Austria.
authors.forename: Annamarie
authors.initials: A
authors.lastname: Wiedermair
authors.affiliation: Department of Gynecology and Obstetrics, Medical University of Innsbruck, Innsbruck, Austria.
authors.forename: Julia
authors.initials: J
authors.lastname: Rössler
authors.affiliation: Skin Cancer Unit, German Cancer Research Center (DKFZ), Heidelberg, Germany.|Department of Dermatology, Venereology and Allergology, University Medical Center Mannheim, Ruprecht-Karl University of Heidelberg, Mannheim, Germany.
authors.forename: Peter
authors.initials: P
authors.lastname: Altevogt
authors.affiliation: Department of Gynecology and Obstetrics, Medical University of Innsbruck, Innsbruck, Austria.
authors.forename: Christian
authors.initials: C
authors.lastname: Marth
authors.affiliation: Department of Gynecology and Obstetrics, Medical University of Innsbruck, Innsbruck, Austria.
authors.forename: Alain Gustave
authors.initials: AG
authors.lastname: Zeimet
mesh_terms: D000368:Aged; D016022:Case-Control Studies; D019175:DNA Methylation; D016889:Endometrial Neoplasms / Q000378:metabolism*; D004717:Endometrium / Q000378:metabolism; D005260:Female; D005500:Follow-Up Studies; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D007150:Immunohistochemistry; D035683:MicroRNAs / Q000378:metabolism*; D008875:Middle Aged; D039842:Neural Cell Adhesion Molecule L1 / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D060888:Real-Time Polymerase Chain Reaction; D015203:Reproducibility of Results; D018570:Risk Assessment
chemical_list: C522425:MIRN34 microRNA, human; D035683:MicroRNAs; D039842:Neural Cell Adhesion Molecule L1
keywords: L1CAM; endometrial cancer; methylation; outcome; qRT-PCR