The Inhibitory Effect Of MG132 On Proliferation, Migration And Differentiation Of HLEC

Objective To study the inhibitory effect of MG132 on proliferation, migration and differentiation of HLEC in vitro, for evaluating the mechanism of MG132 on the prevention of after cataract.Methods1. Human lens anterior capsule was cultured in vitro with explant culture method, and the cultured cells were identified by morphological detection and the characteristic lens-like spherule of HLEC.2. Determined by MTT:The passage HLEC was treated by FGF-2(10ng/ml), MG132(10μM), MG132+FGF-2(FGF-2,10ng/ml;MG132,10μM) and established the control group. MTT method was used to detect the OD values after 24h, and the proliferation rate was calculated.3. Migration ability determination:The passage HLEC was treated by FGF-2 (10ng/ml), MG132(10μM), MG132+FGF-2(FGF-2,10ng/ml;MG132,10μM) and established the control group. A bare area was made by a sterile cotton swab in the cell layer. Count the number of cell in the bare area after 24h and the migration ability was calculated.4. Cell immunohistochemical method:The passage HLEC was treated by TGF-β2 (10ng/ml);MG132(10μM);MG132+TGF-β2(TGF-β2,10ng/ml;MG132,10μM) and established the control group. The expression of FN in the HLEC cytoplasm were traced by cell immunohistochemical method after 24h.Results1. A few of epithelial cells moved from lens capsule in the primary cell culture in the 24h after HLEC was implanted and appeared adherent growth. During long-term culture it can be observed a small ball shape of concentric circles, that was the characteristic lens-like bodys of HLEC in vitro.2.10ng/mlFGF-2 can induce HLEC proliferation 24.2%; 10μM MG132 can effectively restrain the HLEC proliferation, proliferation rate was-25.4%and can effectively restrain the HLEC proliferation induced by FGF-2,and proliferation rate was-20.1%. The differences of the proliferative rates3.10ng/mlFGF-2 can induce HLEC migration, and the migration ability was 33.6%. 10μM MG132 can effectively restrain the HLEC's migration that was induced by FGF-2. The differences of the migration ability between the group of MG132+FGF-2 and the group of FGF-2 were statistically significant(P<0.05).4. Yellowish brown deposit can be detected in HLEC cytoplasm in the MG132+TGF-β2 groups. The FN protein expression level was obvious descended in the MG132 group(P<0.05), the same as the control group. The FN protein expression of MG132 group has no obvious difference.Conclusions1. This experiment of culturing HLEC in vitro was successfully finished, and obtained high purity cells of HLEC that can be used as the cytology test object of after cataract..2. MG132 can effectively inhibit the proliferation, migration and differentiation of HLEC whether FGF-2 and TGF-β2 were existent.