To study the anticancer effect of chitosan nanoparticles (CSNPs) on a different human cancer cell lines and their possible biomedical application as anticancer, CSNPs were prepared based on the ionic gelation of chitosan with tripolyphosphate anions(TPP), The CSNPs were characterized using UV-VIS spectroscopy , transmission electron microscopy, Fourier transform infrared spectroscopy and zeta analysis. Different human cancer cell lines were used as a model. The cytotoxic effects of the CSNPs on Human breast tumour, human hepatocellular carcinoma, human lung cancer, human colon cancer and human hypopharyngeal were monitored by sulforhodamine B colorimetric assays for cytotoxicity. Treatment of different types of cancer cell lines with different concentrations of 140 nm diameter of CSNPs produce a concentration dependent decrease in cell viability and IC50 reached between 1000 and 2000 μg/ml of CSNPs after 48 h of cell incupation .These investigations implied that CSNPs could be effectively applied to improve the anticancer and nanotheraputic effect on different types of cancer.