title: Doublecortin-like kinase 1 exhibits cancer stem cell-like characteristics in a human colon cancer cell line.
abstract: OBJECTIVE
  Colon cancer stem cells (CSCs) are implicated in colorectal cancer carcinogenesis, metastasis, and therapeutic resistance. The identification of these cells could help to develop novel therapeutic strategies. Doublecortin-like kinase 1 (DCLK1) has been viewed as a marker for gastrointestinal stem cells that fuel the self-renewal process, however others view them as a marker of Tuft cells or as an enteroendocrine subtype. The purpose of this study was to use a colon cancer cell line to identify and characterize the stem-like characteristics of the DCLK1+ cell population.
  
  
  METHODS
  To enrich stem-like cells, HCT116 cells (derived from colon adenocarcinomas) were cultured using serum-free media to form spheres under both normal oxygen and hypoxia condition. DCLK1 transcript expression in the adherent parental cells and spheroids was quantified using quantitative real time reverse transcription- polymerase chain reaction [(q)RT-PCR]. DCLK1 protein expression was determined using flow cytometry. Self-renewal capability from adherent parental cells and spheroids was determined using extreme limiting dilution analysis (ELDA).
  
  
  RESULTS
  Under both normal oxygen and hypoxia condition, the adherent parental cells were composed of cells that express low levels of DCLK1. However, spheroids exhibited an increased frequency of cells expressing DCLK1 on both mRNA and protein levels. Cells derived from spheroids also possess stronger self-renewal capability.
  
  
  CONCLUSIONS
  The higher fraction of DCLK1+ cells exhibited by spheroids and hypoxia reflects the stem-like characteristics of these cells. DCLK1 may represent an ideal marker to study and develop effective strategies to overcome chemo-resistance and relapse of colon cancer.
authors.affiliation: Department of Surgery, Tulane University, New Orleans, LA, United States 70112; ; University of Mississippi Cancer Institute, Jackson, MS, United States 39216.
authors.forename: Lianna
authors.initials: L
authors.lastname: Li
authors.forename: Charles F
authors.initials: CF
authors.lastname: Bellows
keywords: Doublecortin-like kinase 1 (DCLK1); cancer stem cells; colorectal cancer; spheroids; stem cell marker