5-Fluorouracil loaded calcium-zinc-gellan and calcium-zinc-gellan-ethyl cellulose microbeads were successfully prepared by simple ionotropic gelation and oil in water ionotropic gelation technique, respectively. Prepared microbeads were characterized by scanning electron microscopy, Fourier transform infrared spectroscopy and evaluated for particle size, drug content, encapsulation efficiency, drug release and cell cytotoxicity study. Microbeads formed were spherical with rough surface. As concentration of gellan and ethyl cellulose has increased encapsulation efficiency, particle size and sustained drug release effect also increased. The release of 5-fluorouracil from microbeads has followed Hixson Crowell model suggesting the mechanism of drug release as dissolution controlled. Cytotoxicity analysis on HT-29 human colon cancer cell lines indicated that 5-FU loaded gellan gum/gellan in combination with ethyl cellulose microbeads leads to sustained releases of drug and thus delayed apoptosis over a long period of time. The formulation with drug:gellan:ethyl cellulose ratio 2.5:7.5:1 was found to be more effectual in terms of sustained drug release activity in addition to anti-cancer activity.