The effect of centrifugation on circulating mRNA quantitation opens up a new scenario in expression profiling from patients with metastatic colorectal cancer

Sze Chuen Cesar Wong, Brigette B Y Ma, Paul B S Lai, Simon S M Ng, Janet F Y Lee, Edwin P. Hui, Money Y Y Lam, Charles M L Chan, Anthony T C Chan

Research output: Journal article publicationJournal articleAcademic researchpeer-review

6 Citations (Scopus)

Abstract

Objectives: To investigate the effect of centrifugation in circulating mRNA quantitation and to design a new, consistent protocol for it. Design and methods: The concentrations of β-catenin mRNA, P-selectin mRNA, cytokeratin 20 mRNA, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA in the plasma, the mononuclear and red cells (MARC) from metastatic colorectal (CRC) patients whose blood samples were centrifuged with 2 different forces, were measured. A new protocol to measure expression profiles in MARC was studied. Results: When the centrifugal force was increased: (1) the concentrations of all transcripts were dramatically decreased in the plasma samples; (2) only the concentrations of P-selectin mRNA and GAPDH mRNA were significantly increased in the mononuclear cell samples; and (3) their concentrations in the red cells did not change significantly. The new protocol found 9 differential expressed genes. Conclusions: Centrifugal force causes inconsistent mRNA quantitation in the plasma. The new protocol discovered 9 differential expressed genes for metastatic CRC.
Original languageEnglish
Pages (from-to)1277-1284
Number of pages8
JournalClinical Biochemistry
Volume40
Issue number16-17
DOIs
Publication statusPublished - 1 Nov 2007
Externally publishedYes

Keywords

  • Centrifugation
  • Expression profiling
  • Metastatic colorectal cancer
  • mRNA
  • Plasma
  • Red cells

ASJC Scopus subject areas

  • Clinical Biochemistry

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