title: Deregulated expression of cryptochrome genes in human colorectal cancer.
abstract: BACKGROUND
  Circadian disruption and deranged molecular clockworks are involved in carcinogenesis. The cryptochrome genes (CRY1 and CRY2) encode circadian proteins important for the functioning of biological oscillators. Their expression in human colorectal cancer (CRC) and in colon cancer cell lines has not been evaluated so far.
  
  
  METHODS
  We investigated CRY1 and CRY2 expression in fifty CRCs and in the CaCo2, HCT116, HT29, SW480 cell lines.
  
  
  RESULTS
  CRY1 (p = 0.01) and CRY2 (p < 0.0001) expression was significantly changed in tumour tissue, as confirmed in a large independent CRC dataset. In addition, lower CRY1 mRNA levels were observed in patients in the age range of 62-74 years (p = 0.018), in female patients (p = 0.003) and in cancers located at the transverse colon (p = 0.008). Lower CRY2 levels were also associated with cancer location at the transverse colon (p = 0.007). CRC patients displaying CRY1 (p = 0.042) and CRY2 (p = 0.043) expression levels over the median were hallmarked by a poorer survival rate. Survey of selected colon cancer cell lines evidenced variable levels of cryptochrome genes expression and time-dependent changes in their mRNA levels. Moreover, they showed reduced apoptosis, increased proliferation and different response to 5-fluorouracil and oxaliplatin upon CRY1 and CRY2 ectopic expression. The relationship with p53 status came out as an additional layer of regulation: higher CRY1 and CRY2 protein levels coincided with a wild type p53 as in HCT116 cells and this condition only marginally affected the apoptotic and cell proliferation characteristics of the cells upon CRY ectopic expression. Conversely, lower CRY and CRY2 levels as in HT29 and SW480 cells coincided with a mutated p53 and a more robust apoptosis and proliferation upon CRY transfection. Besides, an heterogeneous pattern of ARNTL, WEE and c-MYC expression hallmarked the chosen colon cancer cell lines and likely influenced their phenotypic changes.
  
  
  CONCLUSIONS
  Cryptochrome gene expression is altered in CRC, particularly in elderly subjects, female patients and cancers located at the transverse colon, affecting overall survival. Altered CRY1 and CRY2 expression patterns and the interplay with the genetic landscape in colon cancer cells may underlie phenotypic divergence that could influence disease behavior as well as CRC patients survival and response to chemotherapy.
authors.affiliation: Division of Internal Medicine and Chronobiology Unit, IRCCS Scientific Institute and Regional General Hospital "Casa Sollievo della Sofferenza", San Giovanni Rotondo, FG, Italy. g.mazzoccoli@operapadrepio.it.|Department of Medical Sciences, Division of Internal Medicine and Chronobiology Unit, IRCCS Scientific Institute and Regional General Hospital "Casa Sollievo della Sofferenza", San Giovanni Rotondo, FG, 71013, Italy. g.mazzoccoli@operapadrepio.it.
authors.forename: Gianluigi
authors.initials: G
authors.lastname: Mazzoccoli
authors.affiliation: Department of Sciences and Technologies, University of Sannio, Benevento, Italy.
authors.forename: Tommaso
authors.initials: T
authors.lastname: Colangelo
authors.affiliation: Division of Gastroenterology and Research Laboratory, IRCCS Scientific Institute and Regional General Hospital "Casa Sollievo della Sofferenza", San Giovanni Rotondo, FG, Italy.
authors.forename: Anna
authors.initials: A
authors.lastname: Panza
authors.affiliation: Division of Internal Medicine and Chronobiology Unit, IRCCS Scientific Institute and Regional General Hospital "Casa Sollievo della Sofferenza", San Giovanni Rotondo, FG, Italy.
authors.forename: Rosa
authors.initials: R
authors.lastname: Rubino
authors.affiliation: Division of Internal Medicine and Chronobiology Unit, IRCCS Scientific Institute and Regional General Hospital "Casa Sollievo della Sofferenza", San Giovanni Rotondo, FG, Italy.
authors.forename: Angelo
authors.initials: A
authors.lastname: De Cata
authors.affiliation: Division of Internal Medicine and Chronobiology Unit, IRCCS Scientific Institute and Regional General Hospital "Casa Sollievo della Sofferenza", San Giovanni Rotondo, FG, Italy.
authors.forename: Cristiana
authors.initials: C
authors.lastname: Tiberio
authors.affiliation: Division of Gastroenterology and Research Laboratory, IRCCS Scientific Institute and Regional General Hospital "Casa Sollievo della Sofferenza", San Giovanni Rotondo, FG, Italy.
authors.forename: Maria Rosa
authors.initials: MR
authors.lastname: Valvano
authors.affiliation: Division of Gastroenterology and Research Laboratory, IRCCS Scientific Institute and Regional General Hospital "Casa Sollievo della Sofferenza", San Giovanni Rotondo, FG, Italy.
authors.forename: Valerio
authors.initials: V
authors.lastname: Pazienza
authors.affiliation: Medical Genetics, IRCCS Scientific Institute and Regional General Hospital "Casa Sollievo della Sofferenza", San Giovanni Rotondo, FG, Italy.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Merla
authors.affiliation: Medical Genetics, IRCCS Scientific Institute and Regional General Hospital "Casa Sollievo della Sofferenza", San Giovanni Rotondo, FG, Italy.
authors.forename: Bartolomeo
authors.initials: B
authors.lastname: Augello
authors.affiliation: Oncology-Research Laboratory, IRCCS Scientific Institute and Regional General Hospital "Casa Sollievo della Sofferenza", San Giovanni Rotondo, FG, Italy.
authors.forename: Domenico
authors.initials: D
authors.lastname: Trombetta
authors.affiliation: Department of Biology, University of Bari, Bari, Italy.
authors.forename: Clelia Tiziana
authors.initials: CT
authors.lastname: Storlazzi
authors.affiliation: Department of Biology, University of Bari, Bari, Italy.
authors.forename: Gemma
authors.initials: G
authors.lastname: Macchia
authors.affiliation: Division of Gastroenterology and Research Laboratory, IRCCS Scientific Institute and Regional General Hospital "Casa Sollievo della Sofferenza", San Giovanni Rotondo, FG, Italy.
authors.forename: Annamaria
authors.initials: A
authors.lastname: Gentile
authors.affiliation: Division of Gastroenterology and Research Laboratory, IRCCS Scientific Institute and Regional General Hospital "Casa Sollievo della Sofferenza", San Giovanni Rotondo, FG, Italy.
authors.forename: Francesca
authors.initials: F
authors.lastname: Tavano
authors.affiliation: Division of Internal Medicine and Chronobiology Unit, IRCCS Scientific Institute and Regional General Hospital "Casa Sollievo della Sofferenza", San Giovanni Rotondo, FG, Italy.|Euro-Mediterranean Institute of Sciences and Technology (IEMEST), Palermo, Italy.|School of Science and Technology, Nottingham Trent University, Nottingham, UK.|Division of Medicine, University College London, Institute for Liver and Digestive Health, Royal Free Campus, London, UK.
authors.forename: Manlio
authors.initials: M
authors.lastname: Vinciguerra
authors.affiliation: Division of Abdominal Surgery, IRCCS Scientific Institute and Regional General Hospital "Casa Sollievo della Sofferenza", San Giovanni Rotondo, FG, Italy.
authors.forename: Giovanni
authors.initials: G
authors.lastname: Bisceglia
authors.affiliation: Department of Sciences and Technologies, University of Sannio, Benevento, Italy.
authors.forename: Valeria
authors.initials: V
authors.lastname: Rosato
authors.affiliation: Department of Sciences and Technologies, University of Sannio, Benevento, Italy.
authors.forename: Vittorio
authors.initials: V
authors.lastname: Colantuoni
authors.affiliation: Department of Sciences and Technologies, University of Sannio, Benevento, Italy.
authors.forename: Lina
authors.initials: L
authors.lastname: Sabatino
authors.affiliation: Division of Epidemiology and Health Statistics, IRCCS Scientific Institute and Regional General Hospital "Casa Sollievo della Sofferenza", San Giovanni Rotondo, FG, Italy.
authors.forename: Ada
authors.initials: A
authors.lastname: Piepoli
mesh_terms: D000368:Aged; D000970:Antineoplastic Agents / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects / Q000235:genetics; D002453:Cell Cycle / Q000187:drug effects / Q000235:genetics; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D015179:Colorectal Neoplasms / Q000235:genetics* / Q000473:pathology; D056931:Cryptochromes / Q000235:genetics* / Q000378:metabolism; D005260:Female; D018628:Gene Dosage; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D008297:Male; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D013997:Time Factors; D014162:Transfection
chemical_list: D000970:Antineoplastic Agents; C538963:CRY1 protein, human; C538966:CRY2 protein, human; D056931:Cryptochromes; D012333:RNA, Messenger