title: Ki-67 is a strong prognostic marker of non-small cell lung cancer when tissue heterogeneity is considered.
abstract: BACKGROUND
  Ki-67 expression is a well-established prognostic marker in various cancers. However, Ki-67 expression is also known as being heterogeneous. We investigated the prognostic significance of Ki-67 from the view of staining heterogeneity by the technique of Spiral Array.
  
  
  METHODS
  100 cases of resected lung cancer from Toyama university hospital archive were collected. Spiral Array blocks were generated out of 100 cases using 100 μm thick paraffin sections. Four μm thick sections of the Array block were stained for Ki-67. Staining results in each reel were scored for areas with lowest (LS), highest (HS), and average (AS) expression, exclusively in the cancer cells. Heterogeneity score (HeS) was designed as the difference between HS and LS. The scores were divided into four grades (0-3). Clinical information was collected, and the prognostic significance of Ki-67 was analyzed.
  
  
  RESULTS
  Pathological stage was available for 91 patients (43 stage IA, 22 stage IB, 2 stage IIA, 9 stage IIB, 13 stage IIIA, 1 stage IIIB, and 1 stage IV). The HS of Ki-67 score in non-small cell lung cancer was 3 in 17 cases, 2 in 27 cases, 1 in 28 cases, 0 in 21 cases, and 4 reels were lost. 78 cases had clinical follow up. 74 cases had all the information available and were analyzed for correlation between Ki-67 expression and survival. Cases with score 2 and 3 of HS and HeS showed significant poorer prognosis (both P < 0.001), whereas LS or AS did not show significance. The results were identical when analyzing adenocarcinoma and squamous cell carcinoma, separately. Cox multivariate analysis of Ki-67 showed that HS was an independent risk factor affecting overall survival.
  
  
  CONCLUSIONS
  Ki-67 is a strong prognostic marker for non-small cell lung cancer when the degree of highest staining frequency or heterogeneity is considered.
authors.affiliation: Department of Pathology, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan ; Department of Pathology, Nagasaki University Hospital, Nagasaki, Japan.
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Tabata
authors.affiliation: Department of Pathology, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan ; Department of Pathology, Nagasaki University Hospital, Nagasaki, Japan.
authors.forename: Tomonori
authors.initials: T
authors.lastname: Tanaka
authors.affiliation: Department of Pathology, Nagasaki University Hospital, Nagasaki, Japan.
authors.forename: Tomayoshi
authors.initials: T
authors.lastname: Hayashi
authors.affiliation: Department of Surgical Pathology, Laboratory of Pathology, Toyama, Japan ; Department of Pathology, Toyama University Hospital, 2630 Sugitani, Toyama 930-0194, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Hori
authors.affiliation: Department of Pathology, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan ; Department of Pathology, Nagasaki University Hospital, Nagasaki, Japan.
authors.forename: Sayuri
authors.initials: S
authors.lastname: Nunomura
authors.affiliation: Department of Human Pathology, Field of Oncology, Kagoshima University Graduate School of Medical and Dental Sciences, Kagoshima, Japan.
authors.forename: Suguru
authors.initials: S
authors.lastname: Yonezawa
authors.affiliation: Department of Pathology, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan ; Department of Pathology, Nagasaki University Hospital, Nagasaki, Japan.
authors.forename: Junya
authors.initials: J
authors.lastname: Fukuoka
keywords: Biomarkers; Expression; Lung cancer; Pathology; Tissue heterogeneity; Tissue microarray