Objective: To investigate the effect of 15-deoxy-Delta(12,14)-prostaglandin J(2) (15-d-PGJ(2)) on the anoikis of hepatocellular carcinoma (HC) cells and mechanisms thereof.
Methods: Fibronectin or polyhydroxyethyl methacrylate (poly-HEMA) were coated onto tissue culture plates, cell growth status and morphological changes were detected by optical microscope. DNA fragmentation analysis and Flow cytometry were used to measure cell apoptotic activity. Western blotting analysis was performed to detect the levels of focal adhesion kinase (FAK) and phosphorylated FAK(p-FAK). Furthermore, small interfering RNA (siRNA) was used to suppress FAK expression.
Results: The adhesion rate of the BEL-7402 cells treated with 15-d-PGJ(2) began to decrease 12 h after the treatment, time- and dose-dependently compared with the HC cell control group (all P < 0.05); when the concentration of 15-d-PGJ2 was 20 micromol/L, the adherent cells ratio at 24 h and 48 h later were (66.0 +/- 3.6)% and (35.0 +/- 5.0)% respectively. Anoikis of BEL-7402 cells was observed by flow cytometry and DNA fragmentation analysis. Western blotting showed that the p-FAK level of the BEL-7402 cells treated with 15-d-PGJ2 for 24 h decreased dose-dependently, however, the total FAK protein did not change.
Conclusion: 15-d-PGJ(2) induces anoikis and decreases the phosphorylated FAK expression of the hepatocellular carcinoma cells.