Yang-Yin-Jie-Du decoction overcomes gefitinib resistance in non-small cell lung cancer via down-regulation of the PI3K/Akt signalling pathway

Pharm Biol. 2021 Dec;59(1):1294-1304. doi: 10.1080/13880209.2021.1972122.

Abstract

Context: Yang-Yin-Jie-Du Decoction (YYJDD) was used to improve gefitinib efficacy in our clinical practice, but its mechanism remains unclear.

Objective: This study explored if YYJDD could reverse gefitinib resistance.

Materials and methods: H1975 cells were exposed to control, 10 μM gefitinib, 3.2 mg/mL YYJDD or combination treatment. Cell viability was detected by MTT during 0-96 h. Apoptosis and the PI3K/Akt proteins were tested by flow cytometry and western-blot at 24 h. LY294002 was applied to further determine the role of the PI3K/Akt. 23 BALB/c nude xenograft mice received normal saline (n = 5), 80 mg/kg gefitinib (n = 6), 2.35 g/kg lyophilised powder of YYJDD (n = 6) or combination treatment (n = 6) by gavage for 4 weeks and submitted to TUNEL, immunohistochemistry, and western-blot.

Results: In vitro, gefitinib (IC50: 20.68 ± 2.06 μM) and YYJDD (IC50: 6.6 ± 0.21 mg/mL) acted in a moderate synergistic way. Combination treatment inhibited cell viability from 100% to 25.66%. Compared to gefitinb (33.23 ± 3.99%), cell apoptosis was increased with combination treatment (54.11 ± 7.32%), accompanied by down-regulation of the PI3K/Akt. LY294002 further inhibited cell viability, increased apoptosis, and down-regulated p-Akt/Akt. In vivo, the tumour sizes in the combination group (1165.13 ± 157.79 mm3) were smaller than gefitinib alone (1630.66 ± 208.30 mm3). The positive rate of TUNEL staining was increased by combination treatment (22.33 ± 2.75%) versus gefitinib (7.37 ± 0.87%), while the PI3K/Akt was down-regulated.

Discussion and conclusion: YYJDD has potential to overcome gefitinib resistance. Future investigations should be focussed on its specific targets.

Keywords: Chinese herbal medicine; drug-resistant; epidermal growth factor receptor-tyrosine kinase inhibitor; lung neoplasm.

MeSH terms

  • Animals
  • Antineoplastic Agents / pharmacology
  • Apoptosis / drug effects
  • Carcinoma, Non-Small-Cell Lung / drug therapy*
  • Cell Line, Tumor
  • Cell Survival / drug effects
  • Down-Regulation / drug effects
  • Drug Resistance, Neoplasm / drug effects
  • Drugs, Chinese Herbal / pharmacology*
  • Gefitinib / pharmacology*
  • Humans
  • Lung Neoplasms / drug therapy*
  • Mice
  • Mice, Inbred BALB C
  • Mice, Nude
  • Phosphatidylinositol 3-Kinase / metabolism
  • Proto-Oncogene Proteins c-akt / metabolism
  • Signal Transduction / drug effects
  • Xenograft Model Antitumor Assays

Substances

  • Antineoplastic Agents
  • Drugs, Chinese Herbal
  • Phosphatidylinositol 3-Kinase
  • Proto-Oncogene Proteins c-akt
  • Gefitinib

Grants and funding

This work was supported by the National Natural Science Foundation of China under Grant [number: 81704013]; Clinical (gastric cancer) Cooperation Pilot Project of Chinese and Western Medicine for Major and Difficult Diseases; The Key Research and Development Projects of Science and Technology Department of Zhejiang Province under Grant [number: 2015C03G2120026]; and Zhejiang Traditional Chinese Medicine of Science and Technology Program under Grant [number: 2020ZA053].