Silencing of solute carrier family 13 member 5 disrupts energy homeostasis and inhibits proliferation of human hepatocarcinoma cells

J Biol Chem. 2017 Aug 18;292(33):13890-13901. doi: 10.1074/jbc.M117.783860. Epub 2017 Jun 27.

Abstract

The solute carrier family 13 member 5 (SLC13A5), a sodium-coupled citrate transporter, plays a key role in importing citrate from the circulation into liver cells. Recent evidence has revealed that SLC13A5 deletion protects mice from high-fat diet-induced hepatic steatosis and that mutation of the SLC13A5 orthologues in Drosophila melanogaster and Caenorhabditis elegans promotes longevity. However, despite the emerging importance of SLC13A5 in energy homeostasis, whether perturbation of SLC13A5 affects the metabolism and malignancy of hepatocellular carcinoma is unknown. Here, we sought to determine whether SLC13A5 regulates hepatic energy homeostasis and proliferation of hepatoma cells. RNAi-mediated silencing of SLC13A5 expression in two human hepatoma cell lines, HepG2 and Huh7, profoundly suppressed cell proliferation and colony formation, and induced cell cycle arrest accompanied by increased expression of cyclin-dependent kinase inhibitor p21 and decreased expression of cyclin B1. Furthermore, such suppressive effects were also observed on the growth of HepG2 cell-derived xenografts expressing SLC13A5-shRNA in nude mice. Metabolically, knockdown of SLC13A5 in HepG2 and Huh7 cells was associated with a decrease in intracellular levels of citrate, the ratio of ATP/ADP, phospholipid content, and ATP citrate lyase expression. Moreover, both in vitro and in vivo assays demonstrated that SLC13A5 depletion promotes activation of the AMP-activated protein kinase, which was accompanied by deactivation of oncogenic mechanistic target of rapamycin signaling. Together, our findings expand the role of SLC13A5 from facilitating hepatic energy homeostasis to influencing hepatoma cell proliferation and suggest a potential role of SLC13A5 in the progression of human hepatocellular carcinoma.

Keywords: AMPK; ATP citrate lyase (ACLY); SLC13A5; cell proliferation; citrate; energy metabolism; hepatocellular carcinoma; liver metabolism; transporter.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Cell Line, Tumor
  • Cell Proliferation
  • Energy Metabolism*
  • Female
  • Gene Expression Regulation, Neoplastic
  • Hepatoblastoma / metabolism
  • Hepatoblastoma / pathology
  • Hepatoblastoma / therapy*
  • Humans
  • Liver Neoplasms / metabolism
  • Liver Neoplasms / pathology
  • Liver Neoplasms / therapy*
  • Mice, Nude
  • Neoplasm Proteins / agonists
  • Neoplasm Proteins / antagonists & inhibitors*
  • Neoplasm Proteins / genetics
  • Neoplasm Proteins / metabolism
  • RNA Interference
  • RNA, Small Interfering
  • RNAi Therapeutics*
  • Specific Pathogen-Free Organisms
  • Symporters / antagonists & inhibitors*
  • Symporters / genetics
  • Symporters / metabolism
  • Tumor Burden
  • Tumor Stem Cell Assay
  • Xenograft Model Antitumor Assays

Substances

  • Neoplasm Proteins
  • RNA, Small Interfering
  • SLC13A5 protein, human
  • Symporters