The main objective of the present work was to develop liposomal nano-formulation for 9-cis-retinoic acid using di-stearoylphosphocholin/cholesterol mixture, to characterise and to evaluate its anticancer effect on A549 human lung cancer cell lines. The liposomes were prepared using thin film formulation method and characterization of particle size and shape were carried out employing dynamic light scattering and scanning electron microscopy techniques, respectively. The level of drug entrapment into the liposomes and liposomal stability were analysed using spectrophotometry and expressed in terms of percent entrapment. The level of 9-cis-retinoic acid in treated cells also was assayed using spectrophotometry. In vitro drug release level evaluated using a dialysis bag. The anticancer effect was studied using MTT and trypsin blue assays in human lung cancer cell line. The drug entrapment level achieved was 83.33 %. Viability of cancer cells was significantly reduced after liposomal 9-cis-retinoic acid treatment. From these results it could be concluded that the liposomal 9-cis-retinoic acid was easily taken up by the A549 cells compared to free 9-cis-retinoic acid, which might have enhanced the anticancer activity observed in this study.