The lipid composition of the liposomal membrane plays a crucial role in the physical and pharmacological behaviors of liposomal payloads, and it can be tailored to achieve the desired delivery needs. Thus, the aim of the current study was to develop and optimize paclitaxel (PTX) loaded liposome nanocarrier using a thin-film rehydrating method and studying the influence of different zwitterionic Phosphatidylcholine (PC)  lipids with different acyl chains, on the physicochemical properties of the developed PTX liposomal formulations. The studied PC were dipalmitoyl (DPPC), dimyristoyl (DMPC) and palmitoyl-oleoyl (POPC) PCs, in combination with cholesterol. Several characteristics were monitored, including PTX entrapment efficiency (EE) of liposomes, particle size (PS), polydispersity index (PDI), zeta potential (ZP), PTX release in phosphate buffered saline (PBS) with 0.1 tween^® 80 and in PBS / 10 % fetal bovine serum (FBS) cell culture medium, as well as in vitro antitumoral and therapeutic effects. The results demonstrated that DPPC exhibited superiority in the physicochemical properties when compared with DMPC and POPC. The in vitro cytotoxicity findings indicate that the PTX-liposome would be a promising substitutional approach for Taxol^® as a commercial product with a toxic solvent. The results outcomes indicated that the developed PTX-liposome would be a viable drug delivery system with sustaining effect for management of colorectal cancer cells and further clinical use.