Effect of morphine on proliferation and migration of human hepatocellular carcinoma cells:an in vitro experiment / 中华麻醉学杂志

ABSTRACT

Objective To evaluate the effect of morphine on the proliferation and migration of human hepatocellular carcinoma (HCC) cells in an in vitro experiment.Methods The experiment was performed in 2 parts.Experiment Ⅰ Human HCC cells were inoculated in 6-well plates at a density of 2×105 cells/well (2 ml/well) and divided into 6 groups (n=12 each) using a random number tablecontrol group (C group) and 10,25,50,100 and 200 ng/ml morphine groups (M1-M5 groups).Morphine at the final concentration of 10,25,50,100 and 200 ng/ml was added in M1-M5 groups,respectively.The equal volume of phosphate buffer solution was added in group C.The cells were cultured or incubated for 48 h.The expression of μ1-opioid receptor (MOR1) mRNA was measured by real-time polymerase chain reaction.The expression of MOR1 was detected by Western blot in C and M4 groups.Experiment Ⅱ Human HCC cells were inoculated in 96-well plates (1×103 cells/well) or in Transwell chambers(200 μl) and divided into 2 groups (n=9 each) using a random number tablecontrol group (C group) and morphine group (M group).Morphine was added at the final concentration of 100 ng/ml in group M,and the equal volume of phosphate buffer solution (final volume 100 μl/well) was added in group C.The cells were cultured or incubated for 7 days.The cell proliferation was detected by methyl thiazolyl tetrazolium assay at 1-7 days of incubation or culture.The cell migration was determined by Transwell chamber assay at 30 h of incubation or culture.Results Experiment Ⅰ Compared with group C,the expression of MOR1 mRNA was significantly up-regulated in M1-M5 groups,and the expression of MOR1 was significantly up-regulated in group M4 (P<0.01).Compared with group M4,the expression of MOR1 mRNA was significantly down-regulated in M1-M3 and M5 groups (P<0.01).Experiment Ⅱ Compared with group C,the cell proliferation was significantly enhanced on 4th-7th days of incubation,and the number of cells passing through Transwell chambers was increased in group M (P<0.05 or 0.01).The cell proliferation was gradually enhanced on 4th-7th days of incubation in group M (P<0.05).Conclusion Morphine can promote the proliferation and migration of human HCC cells,and the mechanism is related to up-regulation of MOR1 expression in an in vitro experiment.