Exosomes are biological nanoparticles (30−150 nm) secreted in the extracellular area from all of cells, that mediate intercellular message. Exosomes act as the carriers for numerous proteins, DNAs, RNAs and cell-signaling molecules. Therefore, exosomes secreted by the tumor cells are useful for diagnostic purposes because of their persistent presence in the blood and their provision of genetic cargo similar to those in tumor. Due to the risks of aggressive activity and ambiguity of biological activity in other tissues, the use of exosomes in drug delivery and imaging has been limited. However, their high loading, stability and longer circulation time, excellent targeting, high cell penetration performance, and optimal biodegradability have made them potential agents in targeted cancer treatment. Therefore, in addition to examining methods for isolating and loading exosomes, this paper discusses the applications of exosomes in biological measurement, imaging, and therapeutic activities. Also, this review describes the challenges of using exosomes compared to conventional methods and shows that it is very useful to use them due to less aggressive activities. Finally, this review attempts to provide an appropriate incentive by showing the performance of exosomes in cancer therapy through targeted drug delivery, gene therapy, imaging and diagnosis.