Microbiomes include bacteria, viruses, fungi, and other microbes. The microbiome modulates numerous aspects of host physiology and is critical in the pathophysiology of diseases, including colon cancer. Although gut bacterial pathogenesis has become an emerging area in colon cancer, the multi-kingdom aspect of microbiome has yet to be explored. Similar to the bacterial component of the microbiome, the virome contains certain makeup that varies between individuals. In the current review, we introduce the concepts of microbiome and microbiota, research history, methods for modern microbiome studies, and recent progress of mechanisms responsible for microbiome and virome in colon cancer. Furthermore, we discuss our understanding of microbial metabolites in the disease development and therapy of colon cancer. Finally, the gut microbiota can affect the efficacy and toxicity of cancer therapy. We discuss the challenges and future perspectives in microbiome and colon cancer. Exploring and understanding the mechanisms of microbiome will provide insights into effective approaches in potential prevention of treatment of colon cancer.