Global Proteomic Analysis of Breast Cancer Cell Plasma Membrane Electroporation

摘要

The controlled application of electrical pulses (EP) creates reversible pores in the cell plasma membrane, in a phenomenon called electroporation. Electroporation is increasingly used to deliver drugs, genes and other impermeant molecules into the living cells, with great efficacy. Despite its efficacy, there is a lack of comprehensive understanding of the cellular effects induced by electroporation at the molecular level. Considering the electrical nature of biological cells and processes, we hypothesized that EP application can induce a comprehensive effect on cell, not just on the membrane. For this purpose, we utilized label free quantitative global proteomics to study the effects of EP in triple negative breast cancer (TNBC) cells.We applied eight pulses of 1200V/cm, 100µs at 1Hz, a typical parameter used to deliver chemotherapeutics in clinics and investigated the proteomics effect. Our results indicate that EP application significantly regulated the expression of 759 proteins, compared with no treatment (Ctrl). These proteins were localized in multiple cellular components, such as organelles, extracellular region, cell junction, and the membrane. EP also manifested diverse signaling pathways, which are involved in cellular metabolism to the regulation of apoptosis. Particularly, 27 proteins in the oxidative phosphorylation (OXPHOS) pathway and 15 proteins in the citrate cycle (TCA cycle) were upregulated for EP, indicating an upregulation in cellular respiration pathway of energy production. This data provides a critical preliminary insight into the electroporation mechanism, which can be useful to improve the effects of chemotherapeutics delivered with EP for breast cancer treatment.