Analysis Of Intracellular Nucleoside Triphosphate Levels In Normal And Tumor Cell Lines By High-Performance Liquid Chromatography & Studies On Anti-tumor Effects Of 3-oxo Derivate Of Oleanolic Acid (3-7-1) And Its Mechanisms Of Action

Carcinogenesis is a complicated process induced by the interaction of endogenous and exogenous factors. Unquestionably, a genetic change must have occurred in this process. Nucleotides participate in most biochemical process as activated precursors for DNA and RNA synthesis and as intermediate products in biosynthesis. The metabolic pathway for synthesis of nucleotides and recycling bases and nucleosides to nucleotides is subject to alteration as a normal cell goes through the steps which ultimately yield a cancerous cell. Many anticancer and antiviral drugs interfere with the synthesis of DNA and RNA and/or their precursors, notably dNTP and NTP. Batiuk TD et al reported that guanosine and deoxyguanosine exhibited their toxicity to Jurkat cells through two mechanisms: ATP deletion, causing necrosis, and the accumulation of dGTP, resulting in apoptosis. The determination of nucleotides and deoxynucleotides is of fundamental importance either to evaluate the cell energy state or to evaluate the degree of DNA and RNA biosynthesis. This latter aim is particular relevant in studies involving cell cycle regulation, viral infection of cell, and pharmacological activity of anticancer or antiviral drugs able to interfere with nucleic acid replication. An analytical method for the quantitative measurement of NTP and dNTP pools in cells is therefore required for studying the biological differences between normal and tumor cells and the influence of pharmacologically active agents on DNA and RNA sythesis and regulation.Enzyme assay, bioluminescence, ³¹P nuclear magnetic resonance spectroscopy (NMR), high-performance liquid chromatography (HPLC), high-performance capillary electrophoresis (HPCE) and gas chromatography (GC) can be applied to the determination of nucleotides. ³¹P NMR yields signals representing the sum of all nucleotides with similar chemical shifts. It has a great potential for the monitoring of relative changes in nucleotide pools in vivo. The use of HPLC for determining...