Skip to Content
Merck
  • PRIMA-1Met induces myeloma cell death independent of p53 by impairing the GSH/ROS balance.

PRIMA-1Met induces myeloma cell death independent of p53 by impairing the GSH/ROS balance.

Blood (2014-07-10)
Benoît Tessoulin, Géraldine Descamps, Philippe Moreau, Sophie Maïga, Laurence Lodé, Catherine Godon, Séverine Marionneau-Lambot, Thibauld Oullier, Steven Le Gouill, Martine Amiot, Catherine Pellat-Deceunynck
ABSTRACT

The aim of this study was to assess the efficiency of p53 reactivation and induction of massive apoptosis (PRIMA-1(Met)) in inducing myeloma cell death, using 27 human myeloma cell lines (HMCLs) and 23 primary samples. Measuring the lethal dose (LD50) of HMCLs revealed that HMCLs displayed heterogeneous sensitivity, with an LD50 ranging from 4 μM to more than 200 μM. The sensitivity of HMCLs did not correlate with myeloma genomic heterogeneity or TP53 status, and PRIMA-1(Met) did not induce or increase expression of the p53 target genes CDKN1A or TNFRSF10B/DR5. However, PRIMA-1(Met) increased expression of NOXA in a p53-independent manner, and NOXA silencing decreased PRIMA1(Met)-induced cell death. PRIMA-1(Met) depleted glutathione (GSH) content and induced reactive oxygen species production. The expression of GSH synthetase correlated with PRIMA-1(Met) LD50 values, and we showed that a GSH decrease mediated by GSH synthetase silencing or by and L-buthionine sulphoximine, an irreversible inhibitor of γ-glutamylcysteine synthetase, increased PRIMA-1(Met)-induced cell death and overcame PRIMA-1(Met) resistance. PRIMA-1(Met) (10 μM) induced cell death in 65% of primary cells independent of the presence of del17p; did not increase DR5 expression, arguing against an activation of p53 pathway; and synergized with L-buthionine sulphoximine in all samples. Finally, we showed in mouse TP53(neg) JJN3-xenograft model that PRIMA-1(Met) inhibited myeloma growth and synergized with L-buthionine sulphoximine in vivo.

MATERIALS
Product Number
Brand
Product Description

Acetylcysteine, European Pharmacopoeia (EP) Reference Standard
SKU
Pack Size
Availability
Price
Quantity
USP
Acetylcysteine, United States Pharmacopeia (USP) Reference Standard
SKU
Pack Size
Availability
Price
Quantity
Sigma-Aldrich
L-Glutathione oxidized disodium salt, BioReagent, suitable for cell culture
SKU
Pack Size
Availability
Price
Quantity
Sigma-Aldrich
L-Glutathione oxidized disodium salt, ≥98%, powder
SKU
Pack Size
Availability
Price
Quantity
Sigma-Aldrich
Glutathione reduced ethyl ester, ≥90% (TLC)
SKU
Pack Size
Availability
Price
Quantity
Supelco
Glutathione, Pharmaceutical Secondary Standard; Certified Reference Material
SKU
Pack Size
Availability
Price
Quantity
Glutathione, European Pharmacopoeia (EP) Reference Standard
SKU
Pack Size
Availability
Price
Quantity
Supelco
N-Acetyl-L-cysteine, Pharmaceutical Secondary Standard; Certified Reference Material
SKU
Pack Size
Availability
Price
Quantity
Sigma-Aldrich
DL-Cysteine, technical grade
SKU
Pack Size
Availability
Price
Quantity
Sigma-Aldrich
N-Acetyl-L-cysteine, Sigma Grade, ≥99% (TLC), powder
SKU
Pack Size
Availability
Price
Quantity
Sigma-Aldrich
N-Acetyl-L-cysteine, BioXtra, ≥99% (TLC)
SKU
Pack Size
Availability
Price
Quantity
Sigma-Aldrich
L-Glutathione reduced, BioXtra, ≥98.0%
SKU
Pack Size
Availability
Price
Quantity
Sigma-Aldrich
L-Glutathione reduced, suitable for cell culture, BioReagent, ≥98.0%, powder
SKU
Pack Size
Availability
Price
Quantity
Sigma-Aldrich
L-Glutathione oxidized, BioXtra, ≥98%
SKU
Pack Size
Availability
Price
Quantity
Sigma-Aldrich
L-Glutathione oxidized, lyophilized powder
SKU
Pack Size
Availability
Price
Quantity
Sigma-Aldrich
L-Glutathione oxidized, ≥98% (HPLC)
SKU
Pack Size
Availability
Price
Quantity
Sigma-Aldrich
N-Acetyl-L-cysteine, BioReagent, suitable for cell culture
SKU
Pack Size
Availability
Price
Quantity
Sigma-Aldrich
L-Glutathione reduced, ≥98.0%
SKU
Pack Size
Availability
Price
Quantity
SAFC
L-Glutathione oxidized
SKU
Pack Size
Availability
Price
Quantity