N-glycosylation Profiling of Colorectal Cancer Cell Lines Reveals Association of Fucosylation with Differentiation and Caudal Type Homebox 1 (CDX1)/Villin mRNA Expression

Holst S, Deuss AJ, van Pelt GW, van Vliet SJ, Garcia-Vallejo JJ, Koeleman CA, Deelder AM, Mesker WE, Tollenaar RA, Rombouts Y, Wuhrer M

PubMed Entry: 26537799

Sample Cell type Tissue/location Tumour type Staging Tumour site Grade Differentiation Gender Age Race Culture
C10 Colon epithelial descending colon Adenocarcinoma 2 Primary grade II mod-well Male 71 --- adherent
CaCo2 Colon epithelial Colon Carinoma  2 Primary grade II well Male 72 Caucasian adherent
Co115 Colon epithelial Colon ascendens Adenocarcinoma 3 Primary grade IV poorly Female 77  Caucasian adherent
Colo205_VUmc Colon epithelial Colon Adenocarcinoma (anaplastic) 4 ascitic fluid (metastatic site) grade II moderately Male 70   adherent and suspension
Colo320_VUmc Colon epithelial Colon sigmoid Adenocarcinoma 3 Primary --- undifferentiated Female 58 Caucasian loosely adherent
DLD-1 Colon epithelial Colon sigmoid Adenocarcinoma 3 Primary grade III mod-poorly Male 45 Caucasian adherent
HCT116 Colon epithelial Colon proximal Carcinoma 4 Primary grade IV poorly Male 48 --- adherent
HCT15_Vumc Colon epithelial Colon sigmoid Adenocarcinoma 3 Primary --- --- Male   --- ---
HCT8 Colon epithelial Colon ilocecal Adenocarcinoma 3 --- --- --- Male 67 --- adherent
HT29 Colon epithelial Colon Adenocarcinoma 3 Primary grade I well Female 44 Caucasian adherent
KM12_VUmc --- Colon --- 2 Primary grade III --- --- --- --- ---
LOVO Colon epithelial Colon Adenocarcinoma 3 Lymph node metastasis grade IV --- Male 56 Caucasian adherent
LS174T_VUmc** Colon epithelial Colon Adenocarcinoma 2 Primary grade I well Female 58 Caucasian adherent
LS-180 Colon epithelial Colon Adenocarcinoma 2 Primary grade II mod-well Female 58 Caucasian adherent
LS-411N Colon epithelial Colon cecum Cecal carcinoma 2 Primary grade IV poorly Male 32 Caucasian adherent
RKO_VUmc Colon epithelial Colon Carcinoma --- --- grade IV poorly --- --- --- adherent
SW1116 Colon epithelial Colon Adenocarcinoma 1 Primary   well Male 73 Caucasian adherent
SW1398_VUmc --- --- --- --- --- --- --- --- --- --- ---
SW1463 Colon epithelial Rectum Adenocarcinoma 3 Primary grade II-III --- Female 66 Caucasian adherent
SW48 Colon epithelial Transversum Adenocarcinoma 3 Primary grade IV poorly Female 82 Caucasian adherent
SW480 Colon epithelial Colon Adenocarcinoma 2 Primary grade III-IV moderately Male 51 Caucasian adherent
SW620 Colon epithelial Colon Adenocarcinoma 3 Lymph node metastasis grade IV poorly Male 51  Caucasian adherent
SW948_VUmc Colon epithelial Colon Adenocarcinoma 3 Primary grade III moderately Female 81 Caucasian adherent
T84 Colon epithelial Colon Carcinoma --- Lung metastasis --- --- Male 72 Unkown adherent

Data Processing

  • A mean average spectrum for the total cell line sample spectra was generated using an in-house script.
  • The average spectrum was internally recalibrated using glycan peaks of known composition (refer to supplementary information).
  • MassyTools (version 0.1.5.1) was used for the targeted extraction of area under the curve for each CRC cell line profile using the defined composition list.
  • Glycan compsitions were confirmed by MS/MS.
  • The AUC scaled to a total relative intensity of 100 % for each average spectrum.

Additional Notes

  • Spectra obtained over a mass window of m/z 1000–5000 with ion suppression below m/z 900 for a total of 10,000 shots (1000 Hz laser frequency, 200 shots per raster spot during complete random walk).
  • Tandem mass spectrometry (MALDI-TOF-MS/MS) performed for structural elucidation (TOF/TOF mode).
  • Released N-glycans were derivatized by ethyl esterification adapted from Reiding et al. for discrimination of a2,3/6 Sialic acid linkages.

Information from Methods section