Yonsei Med J.  1995 Dec;36(6):480-486. 10.3349/ymj.1995.36.6.480.

Antisense GLUT1 RNA suppresses the transforming phenotypes of NIH 3T3 cells transformed by N-Ras

Affiliations
  • 1Department of Biochemistry and Molecular Biology, the Institute of Genetic Science, Yonsei University College of Medicine, Seoul, Korea.
  • 2Department of Biochemistry, Wonju College of Medicine, Wonju, Korea.

Abstract

An antisense approach was attempted to investigate the role of antisense GLUT1 RNA in suppressing tumor cell phenotypes using N-ras-transformed NIH 3T3 cells. The established cell line transformed by ras showed typical biological characteristics of cancer cells, such as increased glucose transport, GLUT1 mRNA contents, and the ability to form colonies on the soft agar. In this system, the plasmids (pMAM-GLUT1(rev)) which can transcribe the antisense GLUT1 RNA were transfected and the accompanying changes in the phenotypes of the ras-transformed cells were observed. The expression of antisense GLUT1 RNA by induction with dexamethasone reduced the glucose transport by 30% (1.97 +/- 0.13 nmoles) after 4 min incubation when compared to the non-induction group of transformed cell (2.85 +/- 0.19 nmoles). Also, the number of colonies sized over 50 microns on the soft agar was reduced significantly in the antisense RNA expressing group compared to non-induction group. These results suggest that the expression of antisense GLUT1 RNA reduced the glucose transport and transforming potential in soft agar possibly by hybridization with GLUT1 mRNA in N-ras-transformed NIH 3T3 cells.

Keyword

GLUT1 expression; antisense RNA; ras

MeSH Terms

3T3 Cells/metabolism
Animal
Base Sequence
Cell Line, Transformed
Cell Transformation, Neoplastic/metabolism/*pathology
*Genes, ras
Human
Mice
Molecular Sequence Data
Monosaccharide Transport Proteins/*genetics
Phenotype
RNA, Antisense/*metabolism
Support, Non-U.S. Gov't
Tumor Cells, Cultured/metabolism/pathology
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