Effect of WT1 gene silencing on the tumorigenicity of human glioblastoma multiforme cells

Laboratory investigation

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Object

Wilms tumor 1 (WT1) is overexpressed in many human cancers, including glioblastoma multiforme (GBM). In another study, the authors showed that transient WT1 silencing increases the radiosensitivity of glioma cells. Studies of nonglioma cell lines have demonstrated that WT1 promotes cell proliferation and survival; however, this ability has not been rigorously analyzed in human GBM.

Methods

The authors tested the efficacy of 2 sequences of short hairpin RNA (shRNA) directed against WT1 in U251MG human GBM cells and found that 1 sequence was capable of stably silencing WT1 expression. They then evaluated the effect of WT1 silencing on cellular proliferation, invasion, and in vivo tumor formation.

Results

Stable WT1-shRNA expression significantly decreased the proliferation of U251MG cells in vitro as demonstrated by both an adenosine 5′-triphosphate–based viability assay and tritiated thymidine uptake. Furthermore, stable WT1 silencing caused significantly slower growth after the subcutaneous inoculation of tumor cells in the flanks of athymic nude mice and was associated with an increased latency period.

Conclusions

Data in this study provide proof of the principle that downregulation of WT1 causes decreased tumorigenicity of a GBM cell line in vitro and in vivo and suggest that WT1 is a promising target for novel molecular GBM therapies, perhaps in combination with standard treatment modalities.

Abbreviations used in this paper: ANOVA = analysis of variance; ATP = adenosine 5′-triphosphate; GBM = glioblastoma multiforme; GFP = green fluorescent protein; PBS = phosphate-buffered saline; SDS = sodium dodecyl sulfate; shRNA = short hairpin RNA; WT1 = Wilms tumor 1; [3H]Td = tritiated thymidine.
Article Information

Contributor Notes

Address correspondence to: William C. Broaddus, M.D., Ph.D., Department of Neurosurgery, Virginia Commonwealth University, P.O. Box 980631, Richmond, Virginia 23298-0631. email: wcbroadd@vcu.edu.

© AANS, except where prohibited by US copyright law.

Headings
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