signature=6ace3a18d22c102dfc2968f9b85b3a12,Metabolite changes associated with methionine stress sens...

摘要：

Altered cellular metabolism has gained larger recognition as a hallmark of cancer in recent years. While mechanistic insights into the metabolic changes in cancer are limited, the importance of methionine metabolism in cancer cell proliferation has been known and studied for over 30 years. These early studies describe a "methionine-dependence" or "methionine stress sensitivity" of cancer as a phenomenon where the majority of cancer cells cannot proliferate in growth media in which methionine is replaced by its metabolic precursor homocysteine. Interestingly, non-transformed cells are unaffected by this metabolite replacement. Previously, we identified a cell cycle arrest in cancer cells as a response to methionine stress and a global metabolic shift toward a more oxidative, or quiescent, cell state. Using the breast cancer cell line MDA-MB468, we investigate the metabolic response to methionine stress by mass spectroscopy. As a control, we have derived rare clones from MDA-MB468 cells (referred to as MB468res) that are methionine stress insensitive and have lost their tumorigenic ability for anchorage independent growth. The MB468 and MB468res cell line pair is an ideal model to identify unique metabolic signatures linking methionine dependence and tumorigenicity. To analyze metabolite changes in both cell lines, we employed an untargeted approach and lipid analysis during methionine stress over a 48 hour time period. In order to assess homocysteine flux and related metabolites, cells were cultured with a deuterium labeled homocysteine molecule over a12 hour time period. Untargeted profiling of primary metabolites revealed a significant response to media stress within two hours of the media shift, including metabolites involved in methionine metabolism and related pathways. Interestingly, the majority of metabolite abundances decrease, while S-adenosylmethionine (SAM), the down-stream metabolite of methionine and the major methyl donor of the cell, increases over time. This data suggest a possible global inhibition of SAM use during methionine stress conditions. Lipidomics analysis identified an inverse response to methionine stress by phosphatidylcholine (PC) and phosphotidylethanolamine (PE), in MB468 but not MB468res cells. In MB468 cells, PE levels steadily increase over the 48 hour time course whereas PC, a product of SAM and PE, declines. In contrast, levels of PC and PE increase during methionine stress in MB468res cells. Both PC and PE are prominent phospholipid species in eukaryotic membranes and decreased levels may reflect a cellular defect in using SAM as a methyl donor for lipid biogenesis. Our analysis of homocysteine flux suggest that the majority of homocysteine metabolism in both MB468 and MB468res cells is redirected towards the transsulfuration pathway for glutathione synthesis instead of the remethylation pathway. Although the remethylation pathway is the major source for methionine regeneration, efforts to prevent damage from reactive oxygen species by glutathione may be become priority during methionine stress. Previous studies using fluorescence lifetime imaging microscopy, suggest MB468res cells are able to tolerate a metabolic transition to oxidative phosphorylation unlike MB468 cells. This metabolic transition and response during methionine stress may play a large role in the methionine stress sensitivity of cancer. This study investigates metabolic vulnerabilities of cancer from an unexplored area of methionine metabolism.

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