Triptolide abrogates growth of colon cancer and induces cell cycle arrest by inhibiting transcriptional activation of E2F

Amanda R. Oliveira, Georg Beyer, Rohit Chugh, Steven J. Skube, Kaustav Majumder, Sulagna Banerjee, Veena Sangwan, LIHUA LI, Rajinder K Dawra, Subree Subramanian, Ashok K Saluja, Vikas Dudeja

Research output: Contribution to journalArticle

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Abstract

Despite significant progress in diagnostics and therapeutics, over 50 thousand patients die from colorectal cancer annually. Hence, there is urgent need for new lines of treatment. Triptolide, a natural compound isolated from the Chinese herb Tripterygium wilfordii, is effective against multiple cancers. We have synthesized a water soluble analog of triptolide, named Minnelide, which is currently in phase I trial against pancreatic cancer. The aims of the current study were to evaluate whether triptolide/Minnelide is effective against colorectal cancer and to elucidate the mechanism by which triptolide induces cell death in colorectal cancer. Efficacy of Minnelide was evaluated in subcutaneous xenograft and liver metastasis model of colorectal cancer. For mechanistic studies, colon cancer cell lines HCT116 and HT29 were treated with triptolide and the effect on viability, caspase activation, annexin positivity, lactate dehydrogenase release, and cell cycle progression was evaluated. Effect of triptolide on E2F transcriptional activity, mRNA levels of E2F-dependent genes, E2F1- retinoblastoma protein (Rb) binding, and proteins levels of regulator of G1-S transition was also measured. DNA binding of E2F1 was evaluated by chromatin immunoprecipitation assay. Triptolide decreased colon cancer cell viability in a dose- and time-dependent fashion. Minnelide markedly inhibited the growth of colon cancer in the xenograft and liver metastasis model of colon cancer and more than doubles the median survival of animals with liver metastases from colon cancer. Mechanistically, we demonstrate that at low concentrations triptolide induces apoptotic cell death but at higher concentrations it induces cell cycle arrest. Our data suggest that triptolide is able to induce G1 cell cycle arrest by inhibiting transcriptional activation of E2F1. Our data also show that triptolide downregulates E2F activity by potentially modulating events downstream of DNA binding. Therefore, we conclude that Triptolide and Minnelide are effective against colon cancer in multiple pre-clinical models.

Original languageEnglish (US)
Pages (from-to)648-659
Number of pages12
JournalLaboratory Investigation
Volume95
Issue number6
DOIs
StatePublished - Jun 28 2015

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Cell Cycle Checkpoints
Colonic Neoplasms
Transcriptional Activation
Growth
Colorectal Neoplasms
Neoplasm Metastasis
Heterografts
Retinoblastoma Binding Proteins
Liver
triptolide
Cell Death
Tripterygium
G1 Phase Cell Cycle Checkpoints
Annexins
Retinoblastoma Protein
Chromatin Immunoprecipitation
DNA
Caspases
Pancreatic Neoplasms
L-Lactate Dehydrogenase

Cite this

Oliveira, A. R., Beyer, G., Chugh, R., Skube, S. J., Majumder, K., Banerjee, S., ... Dudeja, V. (2015). Triptolide abrogates growth of colon cancer and induces cell cycle arrest by inhibiting transcriptional activation of E2F. Laboratory Investigation, 95(6), 648-659. https://doi.org/10.1038/labinvest.2015.46

Triptolide abrogates growth of colon cancer and induces cell cycle arrest by inhibiting transcriptional activation of E2F. / Oliveira, Amanda R.; Beyer, Georg; Chugh, Rohit; Skube, Steven J.; Majumder, Kaustav; Banerjee, Sulagna; Sangwan, Veena; LI, LIHUA; Dawra, Rajinder K; Subramanian, Subree; Saluja, Ashok K; Dudeja, Vikas.

In: Laboratory Investigation, Vol. 95, No. 6, 28.06.2015, p. 648-659.

Research output: Contribution to journalArticle

Oliveira, AR, Beyer, G, Chugh, R, Skube, SJ, Majumder, K, Banerjee, S, Sangwan, V, LI, LIHUA, Dawra, RK, Subramanian, S, Saluja, AK & Dudeja, V 2015, 'Triptolide abrogates growth of colon cancer and induces cell cycle arrest by inhibiting transcriptional activation of E2F', Laboratory Investigation, vol. 95, no. 6, pp. 648-659. https://doi.org/10.1038/labinvest.2015.46
Oliveira, Amanda R. ; Beyer, Georg ; Chugh, Rohit ; Skube, Steven J. ; Majumder, Kaustav ; Banerjee, Sulagna ; Sangwan, Veena ; LI, LIHUA ; Dawra, Rajinder K ; Subramanian, Subree ; Saluja, Ashok K ; Dudeja, Vikas. / Triptolide abrogates growth of colon cancer and induces cell cycle arrest by inhibiting transcriptional activation of E2F. In: Laboratory Investigation. 2015 ; Vol. 95, No. 6. pp. 648-659.
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