Probing the metabolic phenotype of breast cancer cells by multiple tracer stable isotope resolved metabolomics.
Breast cancers vary by their origin and specific set of genetic lesions, which gives rise to distinct phenotypes and differential response to targeted and untargeted chemotherapies. To explore the functional differences of different breast cell types, we performed Stable Isotope Resolved Metabolomics (SIRM) studies of one primary breast (HMEC) and three breast cancer cells (MCF-7, MDAMB-231, and ZR75-1) having distinct genotypes and growth characteristics, using13C6-glucose,13C-1+2-glucose,13C5,15N2-Gln,13C3-glycerol, and13C8-octanoate as tracers. These tracers were designed to probe the central energy producing and anabolic pathways (glycolysis, pentose phosphate pathway, Krebs Cycle, glutaminolysis, nucleotide synthesis and lipid turnover). We found that glycolysis was not associated with the rate of breast cancer cell proliferation, glutaminolysis did not support lipid synthesis in primary breast or breast cancer cells, but was a major contributor to pyrimidine ring synthesis in all cell types; anaplerotic pyruvate carboxylation was activated in breast cancer versus primary cells. We also found that glucose metabolism in individual breast cancer cell lines differed between in vitro cultures and tumor xenografts, but not the metabolic distinctions between cell lines, which may reflect the influence of tumor architecture/microenvironment.
J.G. Brown Cancer Center, University of Louisville, Louisville, KY, United States; Dept. Chemistry and Center for Regulatory and Environmental Analytical Metabolomics, University of Louisville, Louisville, KY, United States. Electronic address: andrew.lane@uky.edu.
Andrew N
AN
Lane
J.G. Brown Cancer Center, University of Louisville, Louisville, KY, United States. Electronic address: jinlian.tan@louisville.edu.
Julie
J
Tan
J.G. Brown Cancer Center, University of Louisville, Louisville, KY, United States. Electronic address: yali.wang@louisville.edu.
Yali
Y
Wang
J.G. Brown Cancer Center, University of Louisville, Louisville, KY, United States.
Jun
J
Yan
Dept. Chemistry and Center for Regulatory and Environmental Analytical Metabolomics, University of Louisville, Louisville, KY, United States.
Richard M
RM
Higashi
J.G. Brown Cancer Center, University of Louisville, Louisville, KY, United States; Dept. Chemistry and Center for Regulatory and Environmental Analytical Metabolomics, University of Louisville, Louisville, KY, United States. Electronic address: twmfan@gmail.com.
Teresa W-M
TW
Fan
D000818:Animals; D001943:Breast Neoplasms / Q000378:metabolism* / Q000473:pathology; D005260:Female; D064593:Heterografts; D006801:Humans; D007553:Isotope Labeling / Q000379:methods*; D061986:MCF-7 Cells; D053858:Metabolic Networks and Pathways; D055432:Metabolomics / Q000379:methods*; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D009368:Neoplasm Transplantation
(13)C(3)-glycerol; (13)C(3)-glycerol (PubChem CID:753); (13)C(5); (13)C(5),(15)N(2)-Glutamine (PubChem CID:5961); (13)C(6)-glucose; (13)C(6)-glucose (PubChem CID:5793); (13)C(8)-octanoate; (13)C(8)-octanoate (PubChem CID:11939); (15)N(2)-Gln; (PubChem CID:5927); 1D/2D NMR; ATP; Breast cancer metabolism; FT-ICR-MS; Mouse xenografts; Stable isotope-resolved metabolomics; UTP(PubChem CID:6133); alanine (PubChem CID:5950); aspartate (PubChem CID:5460541); glutamate (PubChem CID:5128032); lactate (PubChem CID:91435)