康复学报 (Feb 2023)
Expression and Function of Synthetic Dimeric Exercise-Induced Cytokine Irisin-Irisin
Abstract
ObjectiveStudies have shown that the exercise-induced cytokine Irisin functions were as a dimer, so tandem Irisin-Irisin mock dimers were designed to explore the similarities and differences between recombinant Irisin-Irisin expressed by Escherichia coli and commercial Irisin in activity and function on tumor cells.MethodsEscherichia coli strain BL21 (DE3) and pET-30a expression vectors were selected to obtain recombinant Irisin-Irisin by isopropyl β-D-1-thiogalactopyranoside (IPTG) induction. High purity recombinant Irisin-Irisin was obtained by Ni Sepharose excel. C2C12 skeletal muscle cells were treated with recombinant Irisin-Irisin and monomeric Irisin respectively, and the expressions of mitochondrial related gene UCP-1 were observed to test the activity. The functions were verified in gastric cancer, colorectal cancer and other gastrointestinal tumors, and the activity and function of recombinant Irisin-Irisin and monomeric Irisin on tumor cells were compared.Results(1) Expression and purification of recombinant Irisin-Irisin: compared with the non-induced samples, the recombinant Irisin-Irisin was significantly expressed after IPTG induction; high purity recombinant Irisin-Irisin with a molecular weight of about 27 kDa was obtained by Ni Sepharose excel. (2) Detection of recombinant Irisin-Irisin activity: compared with the blank control group, both the recombinant Irisin-Irisin group and the Irisin group could up-regulate the expression of UCP-1 in C2C12 cells (P<0.05); compared with the Irisin group, the recombinant Irisin-Irisin group had a more significant effect and stronger activity in inducing UCP-1 (P<0.05). (3) Recombinant Irisin-Irisin on tumor function: compared with the blank control group, both the 100 ng/mL and 200 ng/mL Irisin-Irisin groups could inhibit the migration of gastric cancer cells and colorectal cancer cells(P<0.05), but had no effect on their proliferation and clone formation; compared with the Irisin group, the recombinant Irisin-Irisin group had a more significant effect on cell migration inhibition (P<0.05).ConclusionThe recombinant Irisin-Irisin expressed by Escherichia coli has activity, and the activity is higher than that of the monomeric Irisin; recombinant Irisin-Irisin has an effect on the migration of gastric cancer cells and colorectal cancer cells, and the effect was better than that of the monomeric Irisin.
