Aptamers are nucleic acid ligands that are generated by molecular evolution to bind with high affinities and specificities to a large variety of targets, which make them attractive tools to be applied in cancer research. In this review, we highlight the recent progress in aptamer-based applications in cancer molecular research such as cancer targeting, biomarker discovery and therapeutics. Aptamers generated from cell-systematic evolution of ligands by exponential enrichment especially contribute to the discovery of novel membrane proteins as cancer biomarkers. Aptamer-nanoparticle conjugation could achieve higher affinity for cancer detection. Aptamer-conjugated nanocarriers deliver drugs to cancer cells with increased specificity and efficacy, as well as reduced toxicity.