The differentiation therapy is focused on the identification of new agents able to impair the proliferative and metastatic potential of cancer cells through the induction of differentiation. Although several markers of cell differentiation on tumor cells have been identified, their causal relationship with neoplastic competence has not been characterized in sufficient detail to propose their use as new pharmacological targets useful for the design of new differentiation agents. Polyamine level in cancer cells and in body fluids was proposed as potential marker of cell proliferation and differentiation. The main advantage of this marker is the possibility to evaluate the antineoplastic activity of new drugs able to induce cell differentiation and consequently to inhibit tumor growth and metastasis. The presented report shows a simply and highly reproducible reverse-phase high-performance liquid chromatographic (HPLC) method for the determination of ortho-phthalaldehyde (OPA) derivatives of polyamines: putrescine (PUT), cadaverine (CAD), spermidine (SPD) and spermine (SPM). The novelty of this method is the fluorescence response for OPA-derivate of SPM, generally low in other procedures, that has been significantly improved by the use of a fully endcapped packing material with minimal silanol interactions. The limits of detection for PUT, CAD, SPD and SPM were 0.6, 0.7, 0.8, and 0.4 pmol/mL, respectively. The analysis time was ≤ 20 min, and the relative recovery rate was of about 97%. To verify the usefulness of this method, it has been validated in a murine melanoma cell line (B16-F10) treated with two theophylline derivatives (namely 8-chlorotheophylline and 8-bromotheophylline). These two compounds increased the activity of tissue transglutaminase (TG2) and the synthesis of melanin, two recognized markers of melanoma cell differentiation, and significantly reduced the levels of intracellular polyamines.
Chung SI, Folk JE (1972) Transglutaminase from hair follicle of guinea pig. Proc Natl Acad Sci (Wash) 69:303–307CrossRefGoogle Scholar
Corbin JL, Marsh BH, Peters GA (1989) An improved method for analysis of polyamines in plant tissue by precolumn derivatization with o-phthalaldehyde and separation by high performance liquid chromatography. Plant Physiol 90(2):434–439CrossRefGoogle Scholar
Cordella M, Tabolacci C, Senatore C, Rossi S, Mueller S, Lintas C, Eramo A, D’Arcangelo D, Valitutti S, Facchiano A, Facchiano F (2019) Theophylline induces differentiation and modulates cytoskeleton dynamics and cytokines secretion in human melanoma-initiating cells. Life Sci 230:121–131. https://doi.org/10.1016/j.lfs.2019.05.050CrossRefPubMedGoogle Scholar
Lentini A, Vidal-Vanaclocha F, Facchiano F, Caraglia M, Abbruzzese A, Beninati S (2000) Theophylline administration markedly reduces hepatic and pulmonary implantation of B16-F10 melanoma cells in mice. Melanoma Res 10(5):435–443CrossRefGoogle Scholar
Piacentini M, Martinet N, Beninati S, Folk JE (1988) Free and protein-conjugated polyamines in mouse epidermal cells. Effect of high calcium and retinoic acid. J Biol Chem 263(8):3790–3794PubMedGoogle Scholar
Shah U, Kavad M, Raval M (2014) Development and validation of stability-indicating RP-HPLC method for estimation of pamabrom in tablets. Indian J Pharm Sci 76(3):198–202PubMedPubMedCentralGoogle Scholar
Venza M, Visalli M, Cicciu D, Teti D (2001) Determination of polyamines in human saliva by high-performance liquid chromatography with fluorescence detection. J Chromatogr B Biomed Sci Appl 757(1):111–117CrossRefGoogle Scholar