| Objective To study the effect of genistein on the proliferation of rabbit conjunctival fibroblasts (RCF) in vitro . Method RCF cultured in vitro.The third to fifth passage of RCF were investigated for the inhibiting effects of genistein: ⑴ Fibroblasts were placed in 96-well culture plates and incubated for 24 hours, then treated with various concentrations of genistein (0.3125-40μg/ml),5-Fu (10-2-102) and genistein (5,10μg/ml) combination with 5-Fu(1μg/ml).Cultured RCF were investigated by MTT assay after exposure to the drugs 48 hours.⑵RCF were inoculated as method ⑴, treated with genistein (0.625-40μg/ml) for 1,3,5,7days,and investigated by MTT assay respectively. ⑶After RCF being incubating for 24 hours and then treated with genistein (0,5,10,20μg/ml) for 48 hours,the changes of cell cycle were measured by flow cytometer. ⑷At the same time the changes of RCF apoptosis were measured by Annexin V-/PI after treated with genistein (0,2.5,10μg/ml) for 48 hours. Result Genistein (2.5,5,10,20,40μg/ml) suppressed the proliferation of RCF in vitro .The rate of inhibition were 27.16%, 34.02%, 44.55%, 55.08% and 58.90% respectively. The inhibition rate showed a dose-dependent and time-dependent tendency after day1, day3, day5 and day7 of exposure to genistein. When genistein (5,10μg/ml) combinated with 5-Fu (1μg/ml), the rate of inhibition were 45.57% and 49.09% respectively. The cells treated with genistein (0,5,10,20μg/ml) was increased in G2-M phase from 6.34% to 9.37%. The cells exposure to genistein (10μg/ml) for 48 hours showed apoptosis.Conclusion Genistein has a significant inhibition effent on the proliferation of RCF and DNA synthesis. The effect is in a dose-dependent and time-dependent tendency. 5-Fu and genistein appears a good synergistic effect on inhibiting proliferation. Genistein can influence the cell cycle and evoke early apoptosis. The experiment provides a scientific basis for selection of drugs to prevent proliferation after glaucoma and other ocular diseases.
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