Selective induction of apoptosis in mutant p53 premalignant and malignant cancer cells by PRIMA-1 through the c-Jun-NH2-kinase pathway
Authors: Yuehua Mao
Paul W Brandt-Rauf
Yin Li
Ann C Williams
Robert L Fine
Journal: Molecular Cancer Therapeutics (2005)
Discipline: CLINICAL MEDICINE
Keywords: PRIMA-1; apoptosis; mutant p53; colorectal cancer; cell cycle
Affiliations: Experimental Therapeutics Program, Division of Medical Oncology, College of Physicians and Surgeons, Columbia University Medical Center, New York, NY 10032, USA.

Abstract:  PRIMA-1 (p53 reactivation and induction of massive apoptosis) is a chemical compound that was originally identified as a selective mutant p53-dependent growth suppressor by screening a library of low-molecular-weight compounds. However, its mechanism of action is unknown. In this study, we examined toxicity of PRIMA-1 to three premalignant human colorectal adenoma cell lines (RG/C2, BR/C1, and AA/C1) and four colorectal carcinoma cell lines (DLD-1, SW480, LOVO, and HCT116) and its mechanism of action. It selectively induced apoptosis only in the mutant p53 premalignant and malignant colon cell lines, but was not toxic to the wild-type p53 premalignant and malignant colon cell lines. Using stable transfectants of temperature-sensitive p53 mutant Ala(143) in null p53 H1299 lung cancer cells, we found that PRIMA-1 induced significantly more apoptosis in cells with mutant p53 conformation (37 degrees C) than the wild-type p53 conformation (32.5 degrees C). Cell cycle analysis indicated that its inhibition of cell growth was correlated with induction of G(2) arrest. Western blot analysis showed PRIMA-1 increased p21 and GADD45 expression selectively in the mutant p53 cells. However, Fas, Bcl-2 family proteins, and caspases were not involved in PRIMA-1-induced cell death. The c-Jun-NH(2)-kinase (JNK) inhibitor SP 600125, but not p38 mitogen-activated protein kinase inhibitor SB 203580 or extracellular signal-regulated kinase inhibitor PD 98059, blocked PRIMA-1-induced apoptosis. Transfection with a dominant-negative phosphorylation mutant JNK, but not a dominant-negative p38 or wild-type JNK, inhibited PRIMA-1-induced cell death, suggesting that the JNK pathway plays an important role in PRIMA-1-induced apoptosis. PRIMA-1 is a highly selective small molecule toxic to p53 mutant cells and may serve as a prototype for the development of new p53-targeting agents for therapy of premalignant and malignant cells. 

Retracted: No

PubPeer comments:
 Fig 1B of this article contains same set of images used in previous paper without proper mentioning this in the text or figure legends (Same as Fig. S4 L,M; Rossi et al., 2013; 10.1016 j.molcel.2013.02.004) ![file] Same can be observed in Fig. 2B compared with Rossi et al., 2013 paper (Supplemental Fig. 4 P Q). No references about the previous work in the figure legends ![file] , 
 Another possible concern is with Fig. 3C: ![file] It states that long and short exposures (ex.) are shown for CDT2. Curiously, in the right panel, itu2019s not possible to observe any difference in the background between short and long ex. ![file] On the other side, in the left panel you can note clear differences in the background between short and long ex. ![file] This might indicate that the same original image was used to represent the short and long ex. in the panel on the right. Could the authors check? , 
 #1 Thank you for pointing out the repetitive use of this representative images illustrating seam cell fusion and gonadal migration patterns in wildtype and dre-1 or dre-1 daf-12 mutant animals. Unfortunately, we did not notice the use of the same original files as in our previous work (Rossi et al. 2013), which was carried out around that same time as Horn et al. 2014. In both studies the images help to illustrate representative WT and mutant phenotypes in addition to the quantification of these phenotypes from many worms. Phenotypes of dre-1 mutants as depicted here were first described in our independent work from 2007 (Fielenbach et al., Dev Cell). We were able to reproduce these results for the follow-up studies from 2013 and 2014 were we looked for dre-1 suppressors. Both studies use the same experimental setup so that the images align with the linked data. In sum, we should have realized this mistake and cited our previous work also in the respective figure legends. Notably, study results and conclusions are by no means effected. ,