Amyloid precursor protein regulates 5-fluorouracil resistance in human hepatocellular carcinoma cells by inhibiting the mitochondrial apoptotic pathway

J Zhejiang Univ Sci B. 2020;21(3):234-245. doi: 10.1631/jzus.B1900413.

Abstract

Hepatocellular carcinoma (HCC) is a malignant tumor with high morbidity and mortality globally. It accounts for the majority of primary liver cancer cases. Amyloid precursor protein (APP), a cell membrane protein, plays a vital role in the pathogenesis of Alzheimer's disease, and has been found to be implicated in tumor growth and metastasis. Therefore, to understand the relationship between APP and 5-fluorouracil (5-FU) resistance in liver cancer, Cell Counting Kit-8, apoptosis and cell cycle assays, western blotting, and reverse transcription-quantitative polymerase chain reaction (qPCR) analysis were performed. The results demonstrated that APP expression in Bel7402-5-FU cells was significantly up-regulated, as compared with that in Bel7402 cells. Through successful construction of APP-silenced (siAPP) and overexpressed (OE) Bel7402 cell lines, data revealed that the Bel7402-APP751-OE cell line was insensitive, while the Bel7402-siAPP cell line was sensitive to 5-FU in comparison to the matched control group. Furthermore, APP overexpression decreased, while APP silencing increased 5-FU-induced apoptosis in Bel7402 cells. Mechanistically, APP overexpression and silencing can regulate the mitochondrial apoptotic pathway and the expression of apoptotic suppressor genes (B-cell lymphoma-2 (Bcl-2) and B-cell lymphoma-extra large (Bcl-xl)). Taken together, these results preliminarily revealed that APP overexpression contributes to the resistance of liver cancer cells to 5-FU, providing a new perspective for drug resistance.

Keywords: Amyloid precursor protein; 5-Fluorouracil resistance; Mitochondrial apoptotic pathway; Hepatocellular carcinoma.

MeSH terms

  • Amyloid beta-Protein Precursor / physiology*
  • Apoptosis / drug effects*
  • Carcinoma, Hepatocellular / drug therapy*
  • Cell Line, Tumor
  • Drug Resistance, Neoplasm
  • Fluorouracil / pharmacology*
  • Humans
  • Liver Neoplasms / drug therapy*
  • Mitochondria / physiology
  • Proto-Oncogene Proteins c-bcl-2 / genetics
  • bcl-X Protein / genetics

Substances

  • Amyloid beta-Protein Precursor
  • BCL2L1 protein, human
  • Proto-Oncogene Proteins c-bcl-2
  • bcl-X Protein
  • Fluorouracil