Effect Of Hepatocyte Growth Factor On Proliferation And Migration Of Human Lens Epithelial Cells

【Objective】To investigate the effect of hepatocyte growth factor(HGF)on proliferation and migration of cultured human lens epithelial cells(hLECs) and study the possible mechanism of HGF in posterior capsule opacification(PCO) while to explore a new possibility for anti-PCO therapy.【Methods】1.hLECs were cultured and sub-cultured. After identified with the shape and immunohistochemical technique, the cells from the third to fifth generation were used in this experiment. 2.MTT assay was used to detect the proliferation of hLECs:(1)The third generation of hLECs were treated with different concentrations of HGF(2.5,5,10,20,40,50,100μg/L),and the growth status of hLECs was analyzed after 24h using MTT;(2) hLECs were treated with 20μg/L of HGF which induced the maximal proliferation of the cells, and the growth status of hLECs was analyzed after different time(6,12,24,48h) using MTT. 3.FCM was used to detect the cell growth cycle: hLECs were treated with 20μg/L of HGF which induced the maximal proliferation of the cells, and the changes of the growth cycle of the cells were detected after 24h using FCM. 4. an in vitro wound-healing model was used to detect the migration of hLECs: A denuded area was made when hLECs cultured in 6-well-plates were confluent into a monolayer. hLECs were then treated with different concentrations of HGF(10,20,50μg/L),and after 24 hours the cells having migrated into the denuded area were enumerated under light microscope. 5. All figures were transformed to data by HMIAS-2000 system and analyzed by SPSS13.0 to study the effect of HGF on proliferation and migration in cultured hLECs.【Result】1. According to growth characteristics, morphology features and staining features (vimentin and keratin) determine cells were hLECs. 2.MTT assay:The proliferation number of hLECs was 0.179±0.011,0.211±0.010,0.208±0.007 and 0.180±0.009 respectively at the concentration of 10,20,40 and 50μg/L of HGF, showing a significantly increased proliferation in comparison with the negative control group (0.162±0.028)(P<0.05或P<0.01),and the proliferation rate was respectively 10.5﹪,30.2﹪,28.3﹪,11.1﹪.At the concentration of 20μg/L, HGF induced the maximal increase of proliferation rate(P<0.01)after 24 hours. HGF promoted the growth of hLECs in a dose-and–time dependent manner. 3.FCM analysis: There was significant difference of the cell cycle between the negative control group and the HGF group after 24 hours (P<0.05). Showing that in the HGF group the G0/G1-phase cells decreased while S + G2/M-phase cells increased. 4.The wound-healing model: The number of migrated hLECs was10.583±1.594,13.667±1.080 and 26.750±2.092 respectively at the concentration of 10,20 and 50μg/L of HGF after 24 hours, showing a significantly migration in comparison with the negative control group (8.667±1.080)(P<0.05或P<0.01),and the migration ability was increased by 22.1%,57.7%,208.6% respectively. HGF promoted migration of hLECs in a dose dependent manner.【Conclusion】HGF can induce the proliferation and obvious migration of hLECs, HGF was a mitogen and potent migratory factor of hLECs.