Synthesis of a Fluorescently Labeled Compound for the Detection of Arsenic-induced Apoptotic HL60 Cells

A. Lis Femia, C. Facundo Temprana, M. Silvia Amor, Mariano Grasselli, Silvia del V. Alonso

doi:10.2174/157340612800493584

Abstract

Arsenic compounds have shown medical usefulness since they proved to be effective in causing complete remission of acute promyelocytic leukemia. In this work we obtained a fluorescently labeled arsenic compound that can be used with current fluorescence techniques for basic and applied research, focused on arsenic-induced apoptosis studies. This compound is an arsanilic acid bearing a covalently linked FITC that was chemically synthesized and characterized by fluorescence, UV-Vis, mass and FTIR spectrometry. In addition, we assessed its apoptotic activity as well as its fluorescent labeling properties in HL60 cell line as a leukemia cell model through flow cytometry. We obtained a compound with a 1:1 FITC:arsenic ratio and a 595 m/z, confirming its structure by FTIR. This compound proved to be useful at inducing apoptosis in the leukemia cell model and labeling this apoptotic cell population, in such a way that the highest FITC fluorescence correlated with the highest arsenic amount.

Keywords: Apoptosis, arsanilic acid, arsenic compounds, fluorescein, fluorescence techniques, leukemia, spectrometry, FTIR, HL60