At present, the cultivation of cephalopods mainly relies on natural feeds. The absence of artificial compound feeds is a restrictive factor for the large-scale development of the cultivation of cephalopods. The genes related to glucose metabolism in Octopus sinensis, including glucose phosphate isomerase (GPI), phosphoglycerate kinase (PGK), and pyruvate kinase (PK), were cloned, and their expression levels were examined to study the glucose metabolism pattern of O. sinensis and provide a reference for optimizing the proportion of carbohydrates in its compound feed. The results showed that the open reading frame (ORF) length of OsGPI was 1725 bp encoding 574 amino acids, the ORF length of OsPGK was 1248 bp encoding 415 amino acids, and the ORF length of OsPK was 1683 bp encoding 560 amino acids. The sequence alignment and phylogenetic tree results showed that the identity of O. sinensis GPI, PGK, and PK proteins was the closest to that of Octopus bimaculoides. qRT-PCR results showed that these three genes were expressed in different tissues/organs of O. sinensis, with OsGPI having the highest expression level in the digestive gland and brain, and OsPGK and OsPK in the brain, followed by posterior salivary glands and the digestive glands. This difference in expression distribution may be related to the biological functions performed by different tissues/organs. OsGPI, OsPGK and OsPK were expressed in different developmental stages, with the highest expression at the hatchling stage. During starvation stress in larvae, the expression levels of OsGPI, OsPGK, and OsPK decreased significantly after 2 days (d) of starvation, followed by a significant increase in expression levels after 3 d or 4 d of starvation. Finally, the expression levels started to decrease after 5 d of starvation. In the experiment of feeding compound feeds, Charybdis acuta or Sardina pilchardus, the expression levels of OsGPI, OsPGK, and OsPK in the digestive gland of O. sinensis were increased to varying degrees in the compound feed group and S. pilchardus group after 21 d feeding. The expression levels of OsPGK and OsPK decreased to varying degrees. The expression levels of OsGPI, OsPGK, and OsPK were significantly increased in the C. chinensis group after feeding for 21 d and 42 d, compared to those at 0 d. After 42 d of feeding, the expression levels of OsGPI, OsPGK, and OsPK decreased to varying degrees. The expression levels of OsGPI, OsPGK, and OsPK were significantly increased in the C. acuta group after feeding for 21 d and 42 d, compared to those at 0 d. The expression patterns of OsGPI, OsPGK, and OsPK during different embryonic development stages, starvation, and feeding of different feeds indicated that they were involved in regulating glucose metabolism in O. sinensis. These results may aid in the selection of diets and the addition of carbohydrates in compound feeds.