The amino-acid-derived polyamines have been associated with cell growth and cancer and their concentrations in malignant tumors are extremely high compared to those in tissues that are histologically normal. Hence, polyamines have been considered as an interesting object for anticancer therapy. Analysis of polyamine levels in biological fluid can possibly provide helpful information on the types of cancer and progression phase of the disease. Numerous modern analytical methods, including high-performance liquid chromatography, gas chromatography, capillary electrophoresis, and other separation techniques have been widely utilized for analysing polyamine levels. In both tissue culture and experimental animal models, polyamine analogues restrain cell growth and destroy malignant cells. Determination of intracellular analogue contents is also crucial, since analogue accumulation is a causal factor for their antitumour effectiveness. In this review, the latest methods reported for polyamines and polyamine analogs in cancer research are discussed.