Journal of Surgery

ISSN: [Jurnalul de chirurgie]
ISSN: 1584-9341

Open Access

Evaluation of Serum of Breast Cancer Patients using High Resolution Magic Angle Proton Magnetic Resonance Spectroscopy (HR-MAS): A Search for Possible Biomarker?


Kumar S, Ahmed F, Sonkar AA, Saroj JK, Parihaar A, Singhai A and Singh VK

Background: Early diagnosis is an important factor for successful outcome in breast cancer. Current existing prognostic and predictive tool like ER, PR and HER2 status have main utility to guide whether a patient should or should not receive adjuvant endocrine or targeted therapy and situation has become more complex after the discovery of genomic tests like oncotype Dx etc. So the need to enhance the understanding of the disease process and treatment response, a hunt for suitable tumor marker is still on.

Sample collection: 5-7 ml fasting peripheral venous blood was withdrawn from newly diagnosed breast cancer patients. The subjects were properly matched in terms of age, sex, dietary habits and other parameters. The blood was centrifuged and resultant supernatant serum was put into the 3 ml ependorf tube and the specimen was immediately snap frozen in liquid nitrogen and was transferred to the HRMAS lab where it was stored at -80ºC.

HRMAS experiment: The collected samples (Malignant 32, Benign 32 and Healthy Control 28) were thawed and subjected to 800 MHz HRMAS spectrometer. The serum samples were recorded in native form using TSP as an internal standard and a coaxial insert. The spectra were acquired using 1D single pulse and Carr-Purcell-Meiboom-Gill (CPMG) pulse sequence with water suppress [PRESET- 90 degree- (δ-180°-δ)n-Aq] with a echo time of 40 ms. The HRMAS findings were correlated with the standard histopathological report.

Results: The 3D scattered PCA score plot of serum (explaining 83% of the total variance showed distinct group separation among the healthy, fibroadenoma and malignant tumor samples. The PLS-DA model generated considering all the three groups showed a predictive ability of 78% which proves that the model was robust enough for group differentiation. The X-loading plot (PC-1 which explained 69% of total variance) of the cases, exhibited positive loadings of lactate, succinate and alanine in serum. When compared with healthy controls, while glucose was found to be down-regulated in almost all the fibroadenoma and malignant cases. Thus, glucose along with lactate appeared to be the major confounders between the two groups viz. healthy and tumor groups.

Conclusion: All malignant tumors showed up regulation of lactic acid, acetate and choline containing compounds while down regulation of glucose and lipids. The study did provided evidence for the clinical use of these identified metabolites. However, future studies involving large sample size using sequential samples should be carried out.


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