Pterygota alata (Roxb.) R.Br. Bark Fraction Induced Intrinsic Apoptotic Pathway in 4T1 Cells by Decreasing Bcl-2 and Inducing Bax Expression

Pak J Biol Sci. 2021 Jan;24(2):172-181. doi: 10.3923/pjbs.2021.172.181.

Abstract

Background and objective: Pterygota alata (Roxb.) R.Br. bark extract has been studied to have cytotoxic activity on 4T1 cells. This study was conducted to determine the cytotoxic activity of several fractions of Pterygota alata (Roxb.) R.Br. bark against 4T1 breast cancer cells and to investigate the most active fractions on Bcl-2 and Bax expressions.

Materials and methods: The bark of Pterygota alata (Roxb.) R.Br. was extracted using 80% methanol and was fractionated into fractions of n-hexane, chloroform, ethyl acetate, n-butanol and insoluble n-butanol with liquid-liquid partition. Cytotoxic tests were performed using the MTT method and expressions of Bax and Bcl-2 on 4T1 breast cancer cells were detected with immunocytochemical staining. Identification of compounds in the most active fraction using GC-MS.

Results: The results showed that the most active fraction was the insoluble fraction of n-butanol (IFB) with an IC50 of 15.14 μg mL-1. IFB also decreases the expression of Bcl-2 and increases the expression of Bax.

Conclusion: It can be concluded that Pterygota alata (Roxb.) R.Br. bark has the potential to be developed for medical use, especially for breast cancer therapy.

Keywords: 4T1; Bax; Bcl-2; Pterygota alata (Roxb.) R. Br.; apoptosis; cytotoxic; fraction.

MeSH terms

  • Animals
  • Antineoplastic Agents, Phytogenic / isolation & purification
  • Antineoplastic Agents, Phytogenic / pharmacology*
  • Apoptosis / drug effects*
  • Breast Neoplasms / drug therapy*
  • Breast Neoplasms / metabolism
  • Breast Neoplasms / pathology
  • Cell Line, Tumor
  • Chlorocebus aethiops
  • Dose-Response Relationship, Drug
  • Female
  • Inhibitory Concentration 50
  • Mice
  • Plant Bark
  • Plant Extracts / isolation & purification
  • Plant Extracts / pharmacology*
  • Proto-Oncogene Proteins c-bcl-2 / metabolism*
  • Pterygota* / chemistry
  • Signal Transduction
  • Vero Cells
  • bcl-2-Associated X Protein / metabolism*

Substances

  • Antineoplastic Agents, Phytogenic
  • Bax protein, mouse
  • Plant Extracts
  • Proto-Oncogene Proteins c-bcl-2
  • bcl-2-Associated X Protein
  • Bcl2 protein, mouse