The Effects Of Fentanyl On Growth And Apoptosis Of Human Hepatocellular Carcinoma BEL-7404Cells

Objiective:This study by observing the impact of different doses of fentanyl on growth and apoptosis of human hepatocellular carcinoma BEL-7404cells and to analyze its possible mechanism, in order to provide clinical guidance for fentanyl in the treatment of cancer patients.Methods:Human hepatocellular carcinoma BEL-7404cells were devided into five groups:control group,5ng/ml fentanyl group,50ng/ml fentanyl group,500ng/ml fentanyl group and5000ng/ml fentanyl group. BEL-7404cells were cultured to logarithmic phase. Randomly divided into5groups,5ng/ml fentanyl group,50ng/ml fentanyl group,500ng/ml fentanyl group and5000ng/ml fentanyl group The cells in fentanyl groups were incubated with5ng/ml,50ng/ml,500ng/ml and5000ng/ml fentanyl and the control cells were incubated without fentanyl. Inverted microscope to observe each group changes in cell morphology.The wound healing assay was used to detect the cell migration. MTT method and Colony formation were employed to evaluate the level of the cell growth and proliferation. The cell cycle progression and apoptosis were assesed by flow cytometry.Rusults:Inverted microscope observation showed that the growth rate of Fen groups were slower than those in control group, And control cells adherent growth,5ng/ml,50ng/ml,500ng/ml and5000ng/ml Fen result in morphological changes of human hepatocellular carcinoma BEL-7404cells such as the cells were smaller, rounded, refractive index enhancement, and other phenomena;The relative migrations of BEL-7404cells which were incubated with5ng/ml,50ng/ml,500ng/ml and5000ng/ml Fen after24h were (41.29±0.937)%、(33.65±2.007)%、(27.63±1.270)%and (19.38±1.563)%,which were less than that of control cells(P<0.05).The OD value of the experimental groups were less than the control group significantly, Fen on the inhibition rate of BEL-7404cells were also increasing, the inhibition rates of BEL-7404cells in5ng/ml,50ng/ml,500ng/ml and5000ng/ml Fen group were6.1%.10.3%.17.5%and21.0%;Clones of human hepatocellular carcinoma cell line BEL-7404in5ng/ml,50ng/ml,500ng/ml and5000ng/ml Fen group were(10.48±0.43)%.(8.06±0.37)%.(6.66＋0.28)%and (4.48±0.45)%, which were fewer than those in control group(P<0.05). In Fen groups,the proportion of MGC-803cells in G0/G1phase was higher and the proportion of cells in S phase was lower than that in control group(P<0.05).The apoptotic rates of BEL-7404cells in5ng/ml,50ng/ml,500ng/ml and5000ng/ml Fen group were (26.27±2.08)%、(28.88±1.06)%、(32.10±1.4)%and (47.27±1.33)%,which were higher than that in control cells(P<0.05). Conclusions:Fen can induce apoptosis of human hepatocellular carcinoma BEL-7404cells and make cell cycle arrest in G0/G1phase and can reduce migration,thus inhibit growth and proliferation of BEL-7404cells.