title: TIF1γ interferes with TGFβ1/SMAD4 signaling to promote poor outcome in operable breast cancer patients.
abstract: BACKGROUND
  The Transforming growth factor β (TGFβ) signaling has a paradoxical role in cancer development and outcome. Besides, the prognostic significance of the TGFβ1, SMAD4 in breast cancer patients is an area of many contradictions. The transcriptional intermediary factor 1γ (TIF1γ) is thought to interact with the TGFβ/SMAD signaling through different mechanisms. Our study aims to define the prognostic significance of TGFβ1, SMAD4 and TIF1γ expression in breast cancer patients and to detect possible interactions among those markers that might affect the outcome.
  
  
  METHODS
  Immunohistochemistry was performed on tissue microarray (TMA) blocks prepared from samples of 248 operable breast cancer patients who presented at Centre Léon Bérard (CLB) between 1998 and 2001. The intensity and the percentage of stained tumor cells were integrated into a single score (0-6) and a cutoff was defined for high or low expression for each marker. Correlation was done between TGFβ1, SMAD4 and TIF1γ expression with the clinico-pathologic parameters using Pearson's chi-square test. Kaplan-Meier method was used to estimate distant metastasis free survival (DMFS), disease free survival (DFS) and overall survival (OS) and the difference between the groups was evaluated with log-rank test.
  
  
  RESULTS
  223 cases were assessable for TIF1γ, 204 for TGFβ1 and 173 for SMAD4. Median age at diagnosis was 55.8 years (range: 27 to 89 years). Tumors were larger than 20 mm in 49.2% and 45.2% had axillary lymph node (LN) metastasis (N1a to N3). 19.4% of the patients had SBR grade I tumors, 46.8% grade II tumors and 33.9% grade III tumors. ER was positive in 85.4%, PR in 75.5% and Her2-neu was over-expressed in 10% of the cases. Nuclear TIF1γ, cytoplasmic TGFβ1, nuclear and cytoplasmic SMAD4 stainings were high in 35.9%, 30.4%, 27.7% and 52.6% respectively. TIF1γ expression was associated with younger age (p=0.006), higher SBR grade (p<0.001), more ER negativity (p=0.035), and tumors larger than 2 cm (p=0.081), while TGFβ1 was not associated with any of the traditional prognostic factors. TGFβ1 expression in tumor cells was a marker of poor prognosis regarding DMFS (HR=2.28; 95% CI: 1.4 to 3.8; p=0.002), DFS (HR=2.00; 95% CI: 1.25 to 3.5; p=0.005) and OS (HR=1.89; 95 % CI: 1.04 to 3.43; p=0.037). TIF1γ expression carried a tendency towards poorer DMFS (p=0.091), DFS (p=0.143) and OS (p=0.091). In the multivariate analysis TGFβ1 remained an independent predictor of shorter DMFS, DFS and OS. Moreover, the prognostic significance of TGFβ1 was more obvious in the TIF1γ high patient subgroup than in the patients with TIF1γ low expression. The subgroup expressing both markers had the worst DMFS (HR=3.2; 95% CI: 1.7 to 5.9; p<0.0001), DFS (HR=3.02; 95 % CI: 1.6 to 5.6; p<0.0001) and OS (HR=2.7; 95 % CI: 1.4 to 5.4; p=0.005).
  
  
  CONCLUSIONS
  There is a crosstalk between the TIF1γ and the TGFβ1/SMAD4 signaling that deteriorates the outcome of operable breast cancer patients and when combined together they can serve as an effective prognostic tool for those patients.
authors.affiliation: Clinical Oncology Department, Faculty of medicine, Cairo University, Cairo, Egypt. loay.kassem@cairocure.com.
authors.forename: Loay
authors.initials: L
authors.lastname: Kassem
authors.affiliation: U1052 Inserm, UMR CNRS 5286. Université de Lyon, Centre de Recherche en Cancérologie de Lyon, Centre Léon Bérard, Lyon, France. mathieu.deygas@etu.univ-lyon1.fr.
authors.forename: Mathieu
authors.initials: M
authors.lastname: Deygas
authors.affiliation: U1052 Inserm, UMR CNRS 5286. Université de Lyon, Centre de Recherche en Cancérologie de Lyon, Centre Léon Bérard, Lyon, France. lfattet@ucsd.edu.
authors.forename: Laurent
authors.initials: L
authors.lastname: Fattet
authors.affiliation: U1052 Inserm, UMR CNRS 5286. Université de Lyon, Centre de Recherche en Cancérologie de Lyon, Centre Léon Bérard, Lyon, France. jonathan.lopez@lyon.unicancer.fr.
authors.forename: Jonathan
authors.initials: J
authors.lastname: Lopez
authors.affiliation: U1052 Inserm, UMR CNRS 5286. Université de Lyon, Centre de Recherche en Cancérologie de Lyon, Centre Léon Bérard, Lyon, France. thilbault.gouvent@lyon.unicancer.fr.
authors.forename: Thibaut
authors.initials: T
authors.lastname: Goulvent
authors.affiliation: Biostatistics Unit, Centre Léon Bérard, Lyon, France. emilie.lavergne@lyon.unicancer.fr.
authors.forename: Emilie
authors.initials: E
authors.lastname: Lavergne
authors.affiliation: Biostatistics Unit, Centre Léon Bérard, Lyon, France. sylvie.chabaud@lyon.unicancer.fr.
authors.forename: Sylvie
authors.initials: S
authors.lastname: Chabaud
authors.affiliation: Department of surgery, Centre Léon Bérard, Lyon, France. nicolas.carrabin@lyon.unicancer.fr.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Carrabin
authors.affiliation: Department of surgery, Centre Léon Bérard, Lyon, France. nicolas.chopin@lyon.unicancer.fr.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Chopin
authors.affiliation: Department of medical oncology, Centre Léon Bérard, Lyon, France. thomas.bachelot@lyon.unicancer.fr.
authors.forename: Thomas
authors.initials: T
authors.lastname: Bachelot
authors.affiliation: U1052 Inserm, UMR CNRS 5286. Université de Lyon, Centre de Recherche en Cancérologie de Lyon, Centre Léon Bérard, Lyon, France. germain.gillet@lyon.unicancer.fr.
authors.forename: Germain
authors.initials: G
authors.lastname: Gillet
authors.affiliation: Department of pathology, Centre Léon Bérard, Lyon, France. isabelle.treilleux@lyon.unicancer.fr.
authors.forename: Isabelle
authors.initials: I
authors.lastname: Treilleux
authors.affiliation: U1052 Inserm, UMR CNRS 5286. Université de Lyon, Centre de Recherche en Cancérologie de Lyon, Centre Léon Bérard, Lyon, France. ruth.rimokh@lyon.unicancer.fr.
authors.forename: Ruth
authors.initials: R
authors.lastname: Rimokh
mesh_terms: D000328:Adult; D000367:Age Factors; D000368:Aged; D000369:Aged, 80 and over; D001943:Breast Neoplasms / Q000737:chemistry* / Q000473:pathology* / Q000601:surgery; D002277:Carcinoma / Q000737:chemistry* / Q000556:secondary / Q000601:surgery; D002467:Cell Nucleus / Q000737:chemistry; D003593:Cytoplasm / Q000737:chemistry; D018572:Disease-Free Survival; D005260:Female; D006801:Humans; D008207:Lymphatic Metastasis; D008875:Middle Aged; D060787:Neoplasm Grading; D009367:Neoplasm Staging; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000032:analysis; D011960:Receptors, Estrogen / Q000032:analysis; D011980:Receptors, Progesterone / Q000032:analysis; D015398:Signal Transduction; D051901:Smad4 Protein / Q000032:analysis*; D015996:Survival Rate; D014157:Transcription Factors / Q000032:analysis*; D053773:Transforming Growth Factor beta1 / Q000032:analysis*; D047368:Tumor Burden
chemical_list: D018719:Erb-b2 Receptor Tyrosine Kinases; D011960:Receptors, Estrogen; D011980:Receptors, Progesterone; D051901:Smad4 Protein; D014157:Transcription Factors; D053773:Transforming Growth Factor beta1; C497796:SMAD4 protein, human; C511956:TRIM33 protein, human; C508053:ERBB2 protein, human