High-throughput measurement of the Tp53 response to anticancer drugs and random compounds using a stably integrated

Taylor A. Sohn, Ravi Bansal, Gloria H. Su, Kathleen M. Murphy, Scott E. Kern

Research output: Contribution to journalArticle

37 Citations (Scopus)

Abstract

Human Tp53 is normally a short-lived protein. Tp53 protein is stabilized and levels are increased in response to a variety of cellular stresses, including those induced by genotoxic anticancer drugs and environmental exposures. To engineer an efficient assay based on this property, we constructed and integrated a Tp53-specific reporter system into human cancer cells, termed p53R cells. We tested a range of conventional chemotherapeutic agents as well as over 16 000 diverse small compounds. Ionizing radiation and two-thirds of conventional chemotherapeutic agents, but only 0.2% of diverse compounds activated Tp53 activity by two-fold or greater, consistent with the presumptive genotoxic activation of Tp53 function. Cytotoxicity was independent of TP53 genetic status when paired, syngeneic wild-type TP53 and TP53-null cells in culture were treated with compounds that activated Tp53. From the unbiased survey of random compounds, Tp53 activation was strongly induced by an analog of AMSA, an investigational anti-cancer agent. Tp53 was also strongly induced by an N-oxide of quinoline and by dabequine, an experimental antimalarial evaluated in humans; dabequine was reported to be negative in other screens of mutagenicity and clastogenicity but carcinogenic in animal studies. Further exploration of antimalarial compounds identified the common medicinals chloroquine, quinacrine, and amodiaquine as Tp53-inducers. Flavonoids are known to have DNA topoisomerase activity, a Tp53-inducing activity that is confirmed in the assay. A reported clinical association of Tp53 immunopositive colorectal cancers with use of the antihypertensive agents was extended by the demonstration of hydralazine and nifedipine as Tp53-inducers. p53R cells represent an efficient Tp53 functional assay to identify chemicals and other agents with interesting biologic properties, including genotoxicity. This assay may have utility in the identification of novel chemotherapeutic agents, as an adjunct in the pharmaceutical optimization of lead compounds, in the exploration of environmental exposures, and in chemical probing of the Tp53 pathway.

Original languageEnglish (US)
Pages (from-to)949-957
Number of pages9
JournalCarcinogenesis
Volume23
Issue number6
StatePublished - 2002
Externally publishedYes

Fingerprint

Environmental Exposure
Antimalarials
Amsacrine
Amodiaquine
Pharmaceutical Preparations
Tumor Suppressor Protein p53
Quinacrine
Hydralazine
Null Lymphocytes
Type I DNA Topoisomerase
Chloroquine
Nifedipine
Ionizing Radiation
Flavonoids
Oxides
Antihypertensive Agents
Colorectal Neoplasms
Neoplasms
Cell Culture Techniques
Proteins

ASJC Scopus subject areas

  • Cancer Research

Cite this

Sohn, T. A., Bansal, R., Su, G. H., Murphy, K. M., & Kern, S. E. (2002). High-throughput measurement of the Tp53 response to anticancer drugs and random compounds using a stably integrated. Carcinogenesis, 23(6), 949-957.

High-throughput measurement of the Tp53 response to anticancer drugs and random compounds using a stably integrated. / Sohn, Taylor A.; Bansal, Ravi; Su, Gloria H.; Murphy, Kathleen M.; Kern, Scott E.

In: Carcinogenesis, Vol. 23, No. 6, 2002, p. 949-957.

Research output: Contribution to journalArticle

Sohn, TA, Bansal, R, Su, GH, Murphy, KM & Kern, SE 2002, 'High-throughput measurement of the Tp53 response to anticancer drugs and random compounds using a stably integrated', Carcinogenesis, vol. 23, no. 6, pp. 949-957.
Sohn, Taylor A. ; Bansal, Ravi ; Su, Gloria H. ; Murphy, Kathleen M. ; Kern, Scott E. / High-throughput measurement of the Tp53 response to anticancer drugs and random compounds using a stably integrated. In: Carcinogenesis. 2002 ; Vol. 23, No. 6. pp. 949-957.
@article{6dcf9785f2014636b777233ee9498d51,
title = "High-throughput measurement of the Tp53 response to anticancer drugs and random compounds using a stably integrated",
abstract = "Human Tp53 is normally a short-lived protein. Tp53 protein is stabilized and levels are increased in response to a variety of cellular stresses, including those induced by genotoxic anticancer drugs and environmental exposures. To engineer an efficient assay based on this property, we constructed and integrated a Tp53-specific reporter system into human cancer cells, termed p53R cells. We tested a range of conventional chemotherapeutic agents as well as over 16 000 diverse small compounds. Ionizing radiation and two-thirds of conventional chemotherapeutic agents, but only 0.2{\%} of diverse compounds activated Tp53 activity by two-fold or greater, consistent with the presumptive genotoxic activation of Tp53 function. Cytotoxicity was independent of TP53 genetic status when paired, syngeneic wild-type TP53 and TP53-null cells in culture were treated with compounds that activated Tp53. From the unbiased survey of random compounds, Tp53 activation was strongly induced by an analog of AMSA, an investigational anti-cancer agent. Tp53 was also strongly induced by an N-oxide of quinoline and by dabequine, an experimental antimalarial evaluated in humans; dabequine was reported to be negative in other screens of mutagenicity and clastogenicity but carcinogenic in animal studies. Further exploration of antimalarial compounds identified the common medicinals chloroquine, quinacrine, and amodiaquine as Tp53-inducers. Flavonoids are known to have DNA topoisomerase activity, a Tp53-inducing activity that is confirmed in the assay. A reported clinical association of Tp53 immunopositive colorectal cancers with use of the antihypertensive agents was extended by the demonstration of hydralazine and nifedipine as Tp53-inducers. p53R cells represent an efficient Tp53 functional assay to identify chemicals and other agents with interesting biologic properties, including genotoxicity. This assay may have utility in the identification of novel chemotherapeutic agents, as an adjunct in the pharmaceutical optimization of lead compounds, in the exploration of environmental exposures, and in chemical probing of the Tp53 pathway.",
author = "Sohn, {Taylor A.} and Ravi Bansal and Su, {Gloria H.} and Murphy, {Kathleen M.} and Kern, {Scott E.}",
year = "2002",
language = "English (US)",
volume = "23",
pages = "949--957",
journal = "Carcinogenesis",
issn = "0143-3334",
publisher = "Oxford University Press",
number = "6",

}

TY - JOUR

T1 - High-throughput measurement of the Tp53 response to anticancer drugs and random compounds using a stably integrated

AU - Sohn, Taylor A.

AU - Bansal, Ravi

AU - Su, Gloria H.

AU - Murphy, Kathleen M.

AU - Kern, Scott E.

PY - 2002

Y1 - 2002

N2 - Human Tp53 is normally a short-lived protein. Tp53 protein is stabilized and levels are increased in response to a variety of cellular stresses, including those induced by genotoxic anticancer drugs and environmental exposures. To engineer an efficient assay based on this property, we constructed and integrated a Tp53-specific reporter system into human cancer cells, termed p53R cells. We tested a range of conventional chemotherapeutic agents as well as over 16 000 diverse small compounds. Ionizing radiation and two-thirds of conventional chemotherapeutic agents, but only 0.2% of diverse compounds activated Tp53 activity by two-fold or greater, consistent with the presumptive genotoxic activation of Tp53 function. Cytotoxicity was independent of TP53 genetic status when paired, syngeneic wild-type TP53 and TP53-null cells in culture were treated with compounds that activated Tp53. From the unbiased survey of random compounds, Tp53 activation was strongly induced by an analog of AMSA, an investigational anti-cancer agent. Tp53 was also strongly induced by an N-oxide of quinoline and by dabequine, an experimental antimalarial evaluated in humans; dabequine was reported to be negative in other screens of mutagenicity and clastogenicity but carcinogenic in animal studies. Further exploration of antimalarial compounds identified the common medicinals chloroquine, quinacrine, and amodiaquine as Tp53-inducers. Flavonoids are known to have DNA topoisomerase activity, a Tp53-inducing activity that is confirmed in the assay. A reported clinical association of Tp53 immunopositive colorectal cancers with use of the antihypertensive agents was extended by the demonstration of hydralazine and nifedipine as Tp53-inducers. p53R cells represent an efficient Tp53 functional assay to identify chemicals and other agents with interesting biologic properties, including genotoxicity. This assay may have utility in the identification of novel chemotherapeutic agents, as an adjunct in the pharmaceutical optimization of lead compounds, in the exploration of environmental exposures, and in chemical probing of the Tp53 pathway.

AB - Human Tp53 is normally a short-lived protein. Tp53 protein is stabilized and levels are increased in response to a variety of cellular stresses, including those induced by genotoxic anticancer drugs and environmental exposures. To engineer an efficient assay based on this property, we constructed and integrated a Tp53-specific reporter system into human cancer cells, termed p53R cells. We tested a range of conventional chemotherapeutic agents as well as over 16 000 diverse small compounds. Ionizing radiation and two-thirds of conventional chemotherapeutic agents, but only 0.2% of diverse compounds activated Tp53 activity by two-fold or greater, consistent with the presumptive genotoxic activation of Tp53 function. Cytotoxicity was independent of TP53 genetic status when paired, syngeneic wild-type TP53 and TP53-null cells in culture were treated with compounds that activated Tp53. From the unbiased survey of random compounds, Tp53 activation was strongly induced by an analog of AMSA, an investigational anti-cancer agent. Tp53 was also strongly induced by an N-oxide of quinoline and by dabequine, an experimental antimalarial evaluated in humans; dabequine was reported to be negative in other screens of mutagenicity and clastogenicity but carcinogenic in animal studies. Further exploration of antimalarial compounds identified the common medicinals chloroquine, quinacrine, and amodiaquine as Tp53-inducers. Flavonoids are known to have DNA topoisomerase activity, a Tp53-inducing activity that is confirmed in the assay. A reported clinical association of Tp53 immunopositive colorectal cancers with use of the antihypertensive agents was extended by the demonstration of hydralazine and nifedipine as Tp53-inducers. p53R cells represent an efficient Tp53 functional assay to identify chemicals and other agents with interesting biologic properties, including genotoxicity. This assay may have utility in the identification of novel chemotherapeutic agents, as an adjunct in the pharmaceutical optimization of lead compounds, in the exploration of environmental exposures, and in chemical probing of the Tp53 pathway.

UR - http://www.scopus.com/inward/record.url?scp=0036317090&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0036317090&partnerID=8YFLogxK

M3 - Article

VL - 23

SP - 949

EP - 957

JO - Carcinogenesis

JF - Carcinogenesis

SN - 0143-3334

IS - 6

ER -