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Monitoring of Intracellular Enzyme Kinetic Characteristics of Peripheral Mononuclear Cells in Breast Cancer Patients

¹ The Biophysical Interdisciplinary Jerome Schottenstein Center for the Research and the Technology of the Cellome, Department of Physics, Bar-Ilan University, Ramat Gan; ² Departments of Pathology and Cytology, and ³ Surgery "A," Assaf Harofeh Medical Center, Zerifin; and ⁴ Department of Surgery, Rabin Medical Center, Petach Tikva, Israel

A new methodology for the detection of functional response of peripheral blood mononuclear cells against breast cancer (BC) antigens was developed. The method is based on cellular enzymatic activity measurements, using a fluorogenic substrate. We used this method to estimate the kinetic activity of lymphocytes derived from cancer patients and healthy donors. The aim of the study was to determine a possible correlation between the basic characteristics (K_m and V_max) of biochemical enzymatic reactions in live peripheral white mononuclear cells and common clinical-pathological characteristics in BC patients. Our method shows that the enzymatic activity, upon interaction with mitogen or tumor antigens, of the peripheral blood cells in BC patients is different from the enzymatic reactions in healthy individuals. This holds true in the early stages, and the difference persists throughout all of the stages of the disease. This difference is manifested, primarily, by an increase in the K_m values after cell incubation with tumor tissue. It was also demonstrated that higher K_m values of tumor tissue-activated peripheral blood mononuclear cells are associated with a better prognostic status of the BC patients (lymph node-negative tumors, hormone receptor preservation, and the absence of Her-2/neu protein overexpression). Thus, the present methodology may serve as an additional criterion for prognosis and monitoring, both in BC patients, and in individuals associated with high cancer risk.

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