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Investigational New Drugs

, Volume 30, Issue 3, pp 1012–1027 | Cite as

A novel synthetic C-1 analogue of 7-deoxypancratistatin induces apoptosis in p53 positive and negative human colorectal cancer cells by targeting the mitochondria: enhancement of activity by tamoxifen

  • Dennis Ma
  • Phillip Tremblay
  • Kevinjeet Mahngar
  • Pardis Akbari-Asl
  • Jonathan Collins
  • Tomas Hudlicky
  • James McNulty
  • Siyaram Pandey
PRECLINICAL STUDIES

Summary

The natural compound pancratistatin (PST), isolated from the Hymenocallis littoralis plant, specifically induces apoptosis in many cancer cell lines. Unlike many other chemotherapeutics, PST is not genotoxic and has minimal adverse effects on non-cancerous cells. However, its availability for preclinical and clinical work is limited due to its low availability in its natural source and difficulties in its chemical synthesis. Several synthetic analogues of 7-deoxypancratistatin with different modifications at C-1 were synthesized and screened for apoptosis inducing activity in human colorectal cancer (CRC) cells. We found that a C-1 acetoxymethyl derivative of 7-deoxypancratistatin, JC-TH-acetate-4 (JCTH-4), was effective in inducing apoptosis in both p53 positive (HCT 116) and p53 negative (HT-29) human CRC cell lines, demonstrating similar efficacy to that of natural PST. JCTH-4 was able to decrease mitochondrial membrane potential (MMP), increase levels of reactive oxygen species in isolated mitochondria, cause release of the apoptogenic factor cytochrome c (Cyto c) from isolated mitochondria, and induce autophagy in HCT 116 and HT-29 cells. Interestingly, when JCTH-4 was administered with tamoxifen (TAM), there was an enhanced effect in apoptosis induction, reactive oxygen species (ROS) production and Cyto c release by isolated mitochondria, and autophagic induction by CRC cells. Minimal toxicity was exhibited by a normal human fetal fibroblast (NFF) and a normal colon fibroblast (CCD-18Co) cell line. Hence, JCTH-4 is a novel compound capable of selectively inducing apoptosis and autophagy in CRC cells alone and in combination with TAM and may serve as a safer and more effective alternative to current cancer therapies.

Keywords

Colorectal cancer Tamoxifen Combination therapy Apoptosis Autophagy 

Abbreviations

ANT

adenine nucleotide translocase

CK

creatine kinase

CRC

colorectal cancer

CypD

cyclophilin D

Cyto c

cytochrome c

ER

estrogen receptor

IC50

half-maximal inhibitory concentration

JCTH-1

JC-TH-acid-1

JCTH-4

JC-TH-acetate-4

HK

hexokinase

LC3

microtubule-associated protein 1 light chain 3

MDC

monodansylcadaverine

MMP

mitochondrial membrane potential

MRC

mitochondrial respiratory chain

mtDNA

mitochondrial DNA

NFF

normal human fetal fibroblast

PBR

peripheral benzodiazepine receptor

PQ

paraquat

PST

pancratistatin

PTP

permeability transition pore

RFU

relative fluorescence units

ROS

reactive oxygen species

SDHA

succinate dehydrogenase subunit A

TAM

tamoxifen

TMRM

tetramethylrhodamine methyl ester

VDAC

voltage-dependent anion channel

Notes

Acknowledgements

This work has been supported by the Knights of Columbus Chapter 9671 (Windsor, Ontario), NSERC, and a CIHR Frederick Banting and Charles Best Canada Graduate Scholarship awarded to Dennis Ma. Thank you to Carly Griffin for providing the pancratistatin results presented in this manuscript. We would also like to thank Colleen Mailloux for the critical review of this manuscript.

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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Dennis Ma
    • 1
  • Phillip Tremblay
    • 1
  • Kevinjeet Mahngar
    • 1
  • Pardis Akbari-Asl
    • 1
  • Jonathan Collins
    • 2
  • Tomas Hudlicky
    • 2
  • James McNulty
    • 3
  • Siyaram Pandey
    • 1
  1. 1.Department of Chemistry and BiochemistryUniversity of WindsorWindsorCanada
  2. 2.Chemistry Department and Centre for BiotechnologyBrock UniversitySt. CatharinesCanada
  3. 3.Department of ChemistryMcMaster UniversityHamiltonCanada

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