Identification of survival genes in human glioblastoma cells by small interfering RNA screening.
Target identification and validation remain difficult steps in the drug discovery process, and uncovering the core genes and pathways that are fundamental for cancer cell survival may facilitate this process. Glioblastoma represents a challenging form of cancer for chemotherapy. Therefore, we assayed 16,560 short interfering RNA (siRNA) aimed at identifying which of the 5520 unique therapeutically targetable gene products were important for the survival of human glioblastoma. We analyzed the viability of T98G glioma cells 96 h after siRNA transfection with two orthogonal statistical methods and identified 55 survival genes that encoded proteases, kinases, and transferases. It is noteworthy that 22% (12/55) of the survival genes were constituents of the 20S and 26S proteasome subunits. An expression survey of a panel of glioma cell lines demonstrated expression of the proteasome component PSMB4, and the validity of the proteasome complex as a target for survival inhibition was confirmed in a series of glioma and nonglioma cell lines by pharmacological inhibition and RNA interference. Biological networks were built with the other survival genes using a protein-protein interaction network, which identified clusters of cellular processes, including protein ubiquitination, purine and pyrimidine metabolism, nucleotide excision repair, and NF-kappaB signaling. The results of this study should broaden our understanding of the core genes and pathways that regulate cell survival; through either small molecule inhibition or RNA interference, we highlight the potential significance of proteasome inhibition.
Department of Pharmacology and Chemical Biology, Biomedical Science Tower 3, 3501 Fifth Avenue, University of Pittsburgh, Pittsburgh, PA 15260, USA.
Nikhil G
NG
Thaker
Fang
F
Zhang
Peter R
PR
McDonald
Tong Ying
TY
Shun
Michael D
MD
Lewen
Ian F
IF
Pollack
John S
JS
Lazo
D000970:Antineoplastic Agents / Q000627:therapeutic use; D015153:Blotting, Western; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000235:genetics; D052138:Genes, Neoplasm / Q000235:genetics*; D055106:Genome-Wide Association Study; D005909:Glioblastoma / Q000188:drug therapy / Q000235:genetics*; D005910:Glioma / Q000188:drug therapy / Q000235:genetics; D006801:Humans; D046988:Proteasome Endopeptidase Complex / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection
D000970:Antineoplastic Agents; D034741:RNA, Small Interfering; C108898:PSMB4 protein, human; D046988:Proteasome Endopeptidase Complex