Lactate and pyruvate levels correlation with lactate dehydrogenase gene expression and glucose consumption in Tamoxifen-resistant MCF-7 cells using capillary electrophoresis with contactless conductivity detection (CE-C4 D)

Electrophoresis. 2022 Feb;43(3):446-455. doi: 10.1002/elps.202100217. Epub 2021 Nov 1.

Abstract

Breast cancer is the second leading cause of cancer death in women after lung cancer. The first-line treatment of metastatic breast cancer in premenopausal women relies on tamoxifen. The development of tamoxifen resistance is not fully understood. In this study, capillary electrophoresis with capacitively coupled contactless conductivity detector was developed to monitor the changes in lactate and pyruvate levels in supernatant media of three models of developed MCF-7 tamoxifen-resistant cells and correlate these metabolites changes with lactate dehydrogenase genes expression and glucose consumption. The electrophoretic separation was achieved under reversed electroosmotic flow conditions. The linear ranges were 0.15-5 and 0.01-1 mM with a correlation coefficient of 0.9966 and 0.9971 and the limits of detection were 0.01 and 0.02 μM for lactate and pyruvate, respectively. Inter- and intrarun accuracy were in the range of 96.88-105.94% with precision (CV, %) of ≤7.35%. The method was completely validated and the results were in agreement with those obtained using the lactate and glucose assay kits. The results revealed a significant increase in both lactate and pyruvate production in the three tamoxifen-resistant MCF-7 cells models compared to control cells. This increase was correlated with the increase of lactate dehydrogenase genes expression and the increase of glucose consumption.

Keywords: Breast cancer; CE-C4D; Cell culture; Lactate; MCF-7; Pyruvate; Tamoxifen resistance.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Breast Neoplasms* / drug therapy
  • Breast Neoplasms* / genetics
  • Electric Conductivity
  • Electrophoresis, Capillary / methods
  • Female
  • Gene Expression
  • Glucose
  • Humans
  • L-Lactate Dehydrogenase
  • Lactic Acid / analysis
  • MCF-7 Cells
  • Pyruvic Acid*
  • Tamoxifen / pharmacology

Substances

  • Tamoxifen
  • Lactic Acid
  • Pyruvic Acid
  • L-Lactate Dehydrogenase
  • Glucose