Metabolomic Impact of Lidocaine on a Triple Negative Breast Cancer Cell Line

Abstract

Background: Metabolomics and onco-anesthesia are two emerging research fields in oncology. Metabolomics (metabolites analysis) is a new diagnostic and prognostic tool that can also be used for predicting the therapeutic or toxic responses to anticancer treatments. Onco-anesthesia studies assess the impact of anesthesia on disease-free and overall survival after cancer surgery. It has been shown that local anesthetics (LA), particularly lidocaine (LIDO), exert antitumor properties both in vitro and in vivo and may alter the biologic fingerprints of cancer cells. As LA are known to impair mitochondrial bioenergetics and byproducts, the aim of the present study was to assess the impact of LIDO on metabolomic profile of a breast cancer cell line. Methods: Breast cancer MDA-MB-231 cells were exposed for 4 h to 0.5 mM LIDO or vehicle (n = 4). The metabolomic fingerprint was characterized by high resolution magic angle spinning NMR spectroscopy (HRMAS). The multivariate technique using the Algorithm to Determine Expected Metabolite Level Alteration (ADEMA) (Cicek et al., PLoS Comput. Biol., 2013, 9, e1002859), based on mutual information to identify expected metabolite level changes with respect to a specific condition, was used to determine the metabolites variations caused by LIDO. Results: LIDO modulates cell metabolites levels. Several pathways, including glutaminolysis, choline, phosphocholine and total choline syntheses were significantly downregulated in the LIDO group. Discussion: This is the first study assessing the impact of LIDO on metabolomic fingerprint of breast cancer cells. Among pathways downregulated by LIDO, many metabolites are reported to be associated with adverse prognosis when present at a high titer in breast cancer patients. These results fit with the antitumor properties of LIDO and suggest its impact on metabolomics profile of cancer cells. These effects of LIDO are of clinical significance because it is widely used for local anesthesia with cutaneous infiltration during percutaneous tumor biopsy. Future in vitro and preclinical studies are necessary to assess whether metabolomics analysis requires modification of local anesthetic techniques during tumor biopsy.

Keywords: anesthesia; cancer progression; cancer surgery; lidocaine; metabolomics; onco-anesthesia; perioperative period.

FIGURE 1

MDA-MB-231 cell viability exposed for 4 h to increased concentrations of lidocaine (from 0.01 to 10 mM) compared to cells exposed to vehicle alone (purified water, CTL). ANOVA: F (5,30) = 28.16; p < .0001 (***: p < .001).

FIGURE 2

1D ¹H CPMG HRMAS spectra of MDA-MB-231 exposed to vehicle (A) or to lidocaine (0.5 mM) (B). Spectra can be compared because they were normalized to the sample weight. Peaks are identified as below: 1-Acetate 2-Alanine 3-Asparagine 4-Aspartate 5-Choline 8-Glutamate 9-Glutamine 10-Glutathione 11-Glycerol 12-Isoleucine 13-Lactate 15-myo-Inositol 17-Phosphocholine 18-Proline 19-Taurine 21-Valine 22-Glycine.

FIGURE 3

Example of 2D ¹H-¹³C HSQC spectrum of MDA-MB-231 cells exposed to lidocaine (0.5 mM for 4 h). Spots are identified as below: 1-Acetate 2-Alanine 3-Asparagine 4-Aspartate 5-Choline 8-Glutamate 9-Glutamine 10-Glutathione 11-Glycerol 12-Isoleucine 13-Lactate 15-myo-Inositol 17-Phosphocholine 18-Proline 19-Taurine 21-Valine 22-Glycine.

FIGURE 4

Metabolomic network of MDA-MB-231 cells exposed to 0.5 mM of lidocaine or vehicle as control for 4 h. Several pathways which promote proliferation, invasion and metastasis (glutaminolysis, choline, phosphocholine and total choline syntheses) were significantly downregulated in lidocaine group. Red, green and blue arrows indicate the decreased, the increased or the unchanged levels of metabolite after exposure to lidocaine compared to control, respectively.