Andrographolide reversed 5-FU resistance in human colorectal cancer by elevating BAX expression

Biochem Pharmacol. 2016 Dec 1:121:8-17. doi: 10.1016/j.bcp.2016.09.024. Epub 2016 Oct 1.

Abstract

5-FU is the first line therapy for colorectal cancer, however, treatment effect is often hampered by the development of drug resistance or toxicity at high doses. Andrographolide is a natural diterpenoid from Andrographis paniculata which has anti-bacterial, anti-antiviral and anti-inflammation activities. In the current study, we test the hypothesis that Andrographolide reverses 5-FU resistance in colorectal cancer and examine the underlying mechanism. In vitro and vivo studies indicated that Andrographolide treatment significantly re-sensitizes HCT116/5-FUR cells (HCT116 cells which are 5-FU resistant) to cytotoxicity of 5-FU. Mechanism analysis showed that Andrographolide/5-FU co-treatment elevated apoptosis level of HCT116/5-FUR cells with highly increased level of BAX. By using biotin-Andrographolide pull down and cellular thermal shift assay, we found out that Andrographolide can directly target to BAX. Andrographolide-BAX interaction prevented BAX degradation, enhancing mitochondria-mediated apoptosis thus reversed 5-FU resistance while BAX silence diminished this effect. Further, by analyzing patient samples who received 5-FU involved chemotherapy, we found that expression level of BAX is correlated with PFS. Our results here provide a novel combination treatment strategy, especially for patients with 5-FU-resistant tumors expressing low level of BAX. Meanwhile, we also proposed that BAX expression may be a predicted and prognosis marker of 5-FU involved chemotherapy.

Keywords: 5-FU-resistant; Andrographolide; BAX; Human colorectal cancer.

MeSH terms

  • Animals
  • Antineoplastic Combined Chemotherapy Protocols / administration & dosage
  • Antineoplastic Combined Chemotherapy Protocols / pharmacology*
  • Antineoplastic Combined Chemotherapy Protocols / therapeutic use
  • Apoptosis / drug effects
  • Cell Survival / drug effects
  • Colorectal Neoplasms / drug therapy*
  • Colorectal Neoplasms / metabolism
  • Colorectal Neoplasms / pathology
  • Diterpenes / administration & dosage
  • Diterpenes / pharmacology*
  • Diterpenes / therapeutic use
  • Drug Resistance, Neoplasm / drug effects*
  • Fluorouracil / administration & dosage
  • Fluorouracil / pharmacology*
  • Fluorouracil / therapeutic use
  • HCT116 Cells
  • Humans
  • Membrane Potential, Mitochondrial / drug effects
  • Mice, Nude
  • Up-Regulation
  • Xenograft Model Antitumor Assays
  • bcl-2-Associated X Protein / genetics
  • bcl-2-Associated X Protein / metabolism*

Substances

  • BAX protein, human
  • Diterpenes
  • bcl-2-Associated X Protein
  • andrographolide
  • Fluorouracil