Aim: To investigate the effects of macrophage migration inhibitory factor (MIF) on proliferation of human gastric cancer MGC-803 cells and expression of cyclin D1 and p27(Kip1) in them, and further determine whether the effects are related to the PI3K/Akt signal transduction pathway.
Methods: Gastric cancer MGC-803 cells were cultured and then treated with 50 microg/L recombinant human MIF (rhMIF) with and without a PI3K inhibitor, LY294002 (25 micromol/L). MTT assay was used to detect the proliferation of MGC-803 cells. Cell cycle was detected by flow cytometry. Expression of cyclin D1 and p27(Kip1) mRNA was by reverse transcription-polymerase chain reaction. Protein expression of phosphorylated Akt (p-Akt), Akt, cyclin D1 and p27(Kip1) was examined by immunocytochemistry and Western blotting.
Results: rhMIF significantly stimulated the proliferation of MGC-803 cells and cell cycle progression from G1 phase to S phase in a concentration- and time-dependent manner. After the MGC-803 cells were treated with rhMIF for 24 h, the expression of cyclin D1 was significantly up-regulated compared with the cells not treated with rhMIF at both mRNA and protein levels (0.97 +/- 0.02 vs 0.74 +/- 0.01, P = 0.002; 0.98 +/- 0.05 vs 0.69 +/- 0.04, P = 0.003). The p27(Kip1) was down-regulated but only statistically significant at the protein level. rhMIF significantly increased the expression of p-Akt, which reached the peak at 30 min, but did not affect the expression of Akt. However, LY294002 inhibited all the effects of rhMIF.
Conclusion: Macrophage MIF increases the proliferation of gastric cancer cells, induces the expression of cyclin D1 at the transcriptional level and inhibits the expression of p27(Kip1) at the post-transcriptional level via the PI3K/Akt pathway.