Alteration of ganglioside composition by stable transfection with antisense vectors against GD3-synthase gene expression

Guichao Zeng, Donna D. Li, Luoyi Gao, Stéphane Birklé, Erhard Bieberich, Akira Tokuda, Robert K Yu

Research output: Contribution to journalArticle

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Abstract

Gangliosides are ubiquitous components of mammalian cells. Their expression is frequently altered in many tumor types. We previously showed that alteration of the ganglioside composition often resulted in changes in cellular morphology and differentiation of cultured cells. In this study, we targeted sialyltransferase gene expression by the antisense knockdown experiment, and the results showed that inhibition of the expression of gangliosides GD3 and O-acetylated GD3 (OAc-GD3) in the neuroblastoma F-11 cells greatly reduced the tumor growth in nude mice. The sense and antisense vectors containing either a 5' end fragment or the entire sequence of the cDNA coding for GD3-synthase were prepared and used in separate experiments to transfect the F-11 cells which express high levels of gangliosides GD3 and OAc-GD3. Single clones were isolated and expanded. Both the activity of the GD3-synthase and the concentrations of GD3 and OAc-GD3 in the antisense- transfected cells were dramatically decreased as a result of transfection with the antisense expression vectors. Further characterization of the antisense-transfected cells showed reduced rates of cell growth and neurite formation and changes in cellular morphology. When the cells were inoculated in athymic nude mice, the tumor growth rate was remarkably suppressed although the tumor incidence was not affected by the altered ganglioside composition. These results indicate that the tumor-associated ganglioside(s) is(are) involved in regulation of tumor growth, probably through the stimulation of angiogenesis of the tumor.

Original languageEnglish (US)
Pages (from-to)8762-8769
Number of pages8
JournalBiochemistry
Volume38
Issue number27
DOIs
StatePublished - Jul 6 1999

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Gangliosides
Gene expression
Transfection
Tumors
Gene Expression
Chemical analysis
Neoplasms
Nude Mice
Growth
Cells
Sialyltransferases
Cell growth
Neurites
Cellular Structures
Neuroblastoma
Cultured Cells
Complementary DNA
Clone Cells
Experiments
Incidence

ASJC Scopus subject areas

  • Biochemistry

Cite this

Alteration of ganglioside composition by stable transfection with antisense vectors against GD3-synthase gene expression. / Zeng, Guichao; Li, Donna D.; Gao, Luoyi; Birklé, Stéphane; Bieberich, Erhard; Tokuda, Akira; Yu, Robert K.

In: Biochemistry, Vol. 38, No. 27, 06.07.1999, p. 8762-8769.

Research output: Contribution to journalArticle

Zeng, Guichao ; Li, Donna D. ; Gao, Luoyi ; Birklé, Stéphane ; Bieberich, Erhard ; Tokuda, Akira ; Yu, Robert K. / Alteration of ganglioside composition by stable transfection with antisense vectors against GD3-synthase gene expression. In: Biochemistry. 1999 ; Vol. 38, No. 27. pp. 8762-8769.
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