The Inhibitory Effects Of Meloxicam On The Growth Of Human Liver Cancer Cell Line HepG2 And Angiogenesis

Objective:To study the inhibitory effect of a COX-2 inhibitor meloxicam on the growth of human liver cancer cell line HepG2 and potential mechanisms,and research its effect on tumor angiogenesis.To explore the value of meloxicam in the chemoprevention and chemotherapy against hepatocellular carcinoma and offer the feasibility and corresponding theory evidence for subsequent animal and clinical experiment. Methods:The effect of meloxicam on the growth of HepG2 cells was evaluated by MTT assay;The expression of proliferating cell nuclear antigen (PCNA) was assessed by immunocytochemistry method; Apotosis index(AI) was counted by terminal deoxynucleotidly transferase-mediated dUTP nick end-labelling(TUNEL) ;The apoptosis rate (AR)was quantified by flow cytometry (FCM);The effect of meloxicam on the expression of VEGF of HepG2 cells was analysised by ELISA assay; The effect of meloxicam on angiogenesis induced by HepG2 cells and its alone effect on angiogenesis were detected with chick chroioallantoic membrane (CAM) model. Reults:(1)The data of MTT showed that meloxicam significantly suppressed the growth of HepG2 cells in time and dose-dependent manner.The live cells decreased gradually with the increasing of drug concentration and disposal time. (2)Meloxicam inhibited the proliferation of HepG2 cells.The expression of PCNA markly decreased after being incubated with meloxicam for 48 hours.(3)The results of TUNEL and FCM confirmed that meloxicam could induce the apoptosis of HepG2 cells.Compared with the control group,the apoptosis index and rate of experimental group had a great increase.Control group:2.13%±0.80 and 2.08%±0.45;The apoptosis index and rate of HepG2 cells treated with 25,50,100 μmol/L meloxicam were 7.41%±2.80,16.95%±2.59,36.71%±5.05 and 5.26%±1.65,11.50%±2.56,27.81%±2.78,respectively.(4)ELISA assay found that meloxicam could inhibit the expression of VEGF of HepG2 cells.The level of VEGF expression in medium supernatant of HepG2 cells descended gradually with the increasing of drug concentration and disposal time.(5)The medium supernatant of HepG2 cells strongly induced angiogenesis on CAM which could be inhibited by 100μmol/L meloxicam.A large number of blood vessels grew scatterly from the center of the vectors.Meloxicam slightly inhibited the angiogenesis of CAM alone.The blood vessels of 100μmol/L meloxicam group were a little lessthan those of the control group. Conclusion: 1.Meloxicam suppresses the growth of HepG2 cells by inhibiting proliferation and inducing apoptosis. 2.Meloxicam inhibits the expression of VEGF of HepG2 cells which may be one of important mechanisms of its anti-angiogenic effect. 3.Meloxicam shows a mild inhibitory effect on the growth of CAM blood vessels and markly restrains the angiogenesis induced by HepG2 cells. 4.It is significant for meloxicam to be studied in the prevention and therapy of liver cancer.COX-2 inhibitors are promised to play an important role in the chemoprevention and chemotherapy against hepatocellular carcinoma.