Genome-wide siRNA screen for modulators of cell death induced by proteasome inhibitor bortezomib.
Multiple pathways have been proposed to explain how proteasome inhibition induces cell death, but mechanisms remain unclear. To approach this issue, we performed a genome-wide siRNA screen to evaluate the genetic determinants that confer sensitivity to bortezomib (Velcade (R); PS-341). This screen identified 100 genes whose knockdown affected lethality to bortezomib and to a structurally diverse set of other proteasome inhibitors. A comparison of three cell lines revealed that 39 of 100 genes were commonly linked to cell death. We causally linked bortezomib-induced cell death to the accumulation of ASF1B, Myc, ODC1, Noxa, BNIP3, Gadd45alpha, p-SMC1A, SREBF1, and p53. Our results suggest that proteasome inhibition promotes cell death primarily by dysregulating Myc and polyamines, interfering with protein translation, and disrupting essential DNA damage repair pathways, leading to programmed cell death.
Chen
Siquan
S
Discovery Technologies, Discovery Oncology Biology, and Medical Biostatistics, Millennium Pharmaceuticals, Inc., Cambridge, MA 02139, USA.
Blank
Jonathan L
JL
Peters
Theodore
T
Liu
Xiaozhen J
XJ
Rappoli
David M
DM
Pickard
Michael D
MD
Menon
Saurabh
S
Yu
Jie
J
Driscoll
Denise L
DL
Lingaraj
Trupti
T
Burkhardt
Anne L
AL
Chen
Wei
W
Garcia
Khristofer
K
Sappal
Darshan S
DS
Gray
Jesse
J
Hales
Paul
P
Leroy
Patrick J
PJ
Ringeling
John
J
Rabino
Claudia
C
Spelman
James J
JJ
Morgenstern
Jay P
JP
Lightcap
Eric S
ES
D000970:Antineoplastic Agents / Q000494:pharmacology*; D001897:Boronic Acids / Q000494:pharmacology*; D000069286:Bortezomib; D016923:Cell Death / Q000187:drug effects* / Q000235:genetics; D003110:Colonic Neoplasms / Q000188:drug therapy / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D004249:DNA Damage; D055785:Gene Knockdown Techniques; D045325:HCT116 Cells; D006367:HeLa Cells; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics; D008545:Melanoma / Q000188:drug therapy / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D011480:Protease Inhibitors / Q000494:pharmacology*; D061988:Proteasome Inhibitors; D017346:Protein Serine-Threonine Kinases / Q000096:biosynthesis / Q000235:genetics; D016271:Proto-Oncogene Proteins c-myc / Q000096:biosynthesis / Q000235:genetics; D011719:Pyrazines / Q000494:pharmacology*; D034741:RNA, Small Interfering / Q000235:genetics*; D012270:Ribosomes / Q000187:drug effects; D058570:TOR Serine-Threonine Kinases; D014162:Transfection
D000970:Antineoplastic Agents; D001897:Boronic Acids; D047908:Intracellular Signaling Peptides and Proteins; C489427:MYC protein, human; D011480:Protease Inhibitors; D061988:Proteasome Inhibitors; D016271:Proto-Oncogene Proteins c-myc; D011719:Pyrazines; D034741:RNA, Small Interfering; D000069286:Bortezomib; C546842:MTOR protein, human; D017346:Protein Serine-Threonine Kinases; D058570:TOR Serine-Threonine Kinases