Our current knowledge of the translational mechanisms and the detailed structural insights of its components have highlighted the characteristically exclusive role of tRNAs and aminoacyl-tRNA synthetase diversity in the evolution of the genetic codes. Phenomenal advancements in mass spectrometry and high-throughput sequencing have enabled the researchers in developing a better understanding of the complex landscape of tRNA modifications. Emerging evidence has started to shed light on linchpin role of tRNAs in carcinogenesis and metastatic dissemination. In this chapter, we have provided an overview of the role of tRNAs, aminoacyl-tRNA synthetases, and tRNA-derived small RNAs in the onset and progression of cancer.