Mass spectrometry analysis shows the biosynthetic pathways supported by pyruvate carboxylase in highly invasive breast cancer cells

BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR BASIS OF DISEASE

卷:1863

Article

Phannasil, Phatchariya¹  Ansari, Israr-ul H.²  El Azzouny, Mahmoud³  Longacre, Melissa J.²  Rattanapornsompong, Khanti¹  Burant, Charles F.³  MacDonald, Michael J.²  Jitrapakdee, Sarawut¹ 

[1] Mahidol Univ, Dept Biochem, Fac Sci, Bangkok, Thailand

[2] Univ Wisconsin, Sch Med & Publ Hlth, Childrens Diabet Ctr, Madison, WI USA

[3] Univ Michigan, Dept Internal Med, Ann Arbor, MI 48109 USA

关键词: Gene knockout;    Pyruvate carboxylase (PC);    Breast cancer;    Cell proliferation rate;    Mitochondrial biosynthesis;    Cancer biology;    Anaplerosis;    Mitochondrial metabolism;   

DOI  :  10.1016/j.bbadis.2016.11.021

来源: Elsevier

【 摘 要 】

We recently showed that the anaplerotic enzyme pyruvate carboxylase (PC) is up-regulated in human breast cancer tissue and its expression is correlated with the late stages of breast cancer and tumor size [Phannasil et al., PloS One 10, e0129848, 2015]. In the current study we showed that PC enzyme activity is much higher in the highly invasive breast cancer cell line MDA-MB-231 than in less invasive breast cancer cell lines. We generated multiple stable PC knockdown cell lines from the MDA-MB-231 cell line and used mass spectrometry with C-13(6)-glucose and C-13(5)-glutamine to discern the pathways that use PC in support of cell growth. Cells with severe PC knockdown showed a marked reduction in viability and proliferation rates suggesting the perturbation of pathways that are involved in cancer invasiveness. Strong PC suppression lowered glucose incorporation into downstream metabolites of oxaloacetate, the product of the PC reaction, including malate, citrate and aspartate. Levels of pyruvate, lactate, the redox partner of pyruvate, and acetyl-CoA were also lower suggesting the impairment of mitochondrial pyruvate cycles. Serine, glycine and 5-carbon sugar levels and flux of glucose into fatty acids were decreased. ATP, ADP and NAD(H) levels were unchanged indicating that PC suppression did not significantly affect mitochondrial energy production. The data indicate that the major metabolic roles of PC in invasive breast cancer are primarily anaplerosis, pyruvate cycling and mitochondrial biosynthesis of precursors of cellular components required for breast cancer cell growth and replication. (C) 2016 Elsevier B.V. All rights reserved.

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