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Glycoconjugate Journal

, Volume 16, Issue 10, pp 659–666 | Cite as

Deletion of A-antigen in a human cancer cell line is associated with reduced promoter activity of CBF/NF-Y binding region, and possibly with enhanced DNA methylation of A transferase promoter

  • Sadahiko Iwamoto
  • Donald A. Withers
  • Kazuko Handa
  • Sen-itiroh Hakomori
Article

Abstract

Employing blood group A and A+ clones derived from the same parental colonic cancer cell lines, we studied the molecular mechanism of deletion/ reduction vs. continuous expression of A antigen in A tumors, a crucial determinant of human tumor malignancy. A transferase mRNA level in one of the A clones (A SW480) was undetectable, while that in A+ SW480 was strongly detectable by semiquantitative RT-PCR. Relatively lower (∼1/3) transcript level was detectable in another A clone (A HT29) in comparison to A+ HT29 by the same RT-PCR procedure, although none of these tumor cell lines showed detectable level of A transcript by Northern blotting or RNase protection methods. Therefore, subsequent studies were performed employing A vs. A+ SW480 clones. Deletion of A transcript in A cells was not due to gene deletion, since Southern blot analysis showed equal presence of genomic DNA regardless of A vs. A+ (SW480 or HT29) or B+ (KATOIII) tumor cells. Two transcriptional control mechanisms leading to differences of A expression in SW480 cells are indicated.

i. Luciferase assay in A and A+ SW480 cells showed that promoter activities of segments of 5′ flanking sequence of ABO gene reflected transcript levels in these cell lines. The enhancing activity of a 43bp tandem repeat unit located between −3899 to −3618 was reduced in A compared to A+ cells.

ii. Distinct differences in the pattern of CpG dinucleotide methylation were found in A vs. A+ cells. Therefore, the methylation process of A promoter DNA may be another important factor controlling A activity in SW480 tumor cells.

Since proliferation and motility of tumor cells are associated closely with A expression, transcription control mechanism for expression of A transferase as described above may be of crucial importance in defining human tumor malignancy.

A− and A+ clones promoter activity CBF/NF-Y binding site CpG island methylation 

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Copyright information

© Kluwer Academic Publishers 1999

Authors and Affiliations

  • Sadahiko Iwamoto
    • 1
    • 2
  • Donald A. Withers
    • 1
  • Kazuko Handa
    • 1
  • Sen-itiroh Hakomori
    • 1
  1. 1.Pacific Northwest Research Institute, and Depts. of Pathobiology and MicrobiologyUniv. of WashingtonSeattleUSA and
  2. 2.Department of Legal Medicine and Human Genetics, Jichi Medical School, Minamikawachi-machiTochigiJapan

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