贝伐单抗对人翼状胬肉成纤维细胞的促凋亡作用研究

Effection of Bevacizumab on the Inhibition of human Pterygium Fibroblasts in vitro

目的观察贝伐单抗(Bevacizumab)对人翼状胬肉成纤维细胞〔雨林木风1〕(Human Pterygial Fibroblast,HPF)的影响,探索其作用机制。方法 MTT法及流式细胞术检测贝伐单抗对HPF细胞的影响;Western blot法检测细胞中caspase-3蛋白表达的变化。结果 MTT示7.5、10、12.5mg.mL-1的贝伐单抗对HPF细胞的生长均有明显抑制(P<0.05),48h时抑制率分别为10.49%、17.03%、26.69%,抑制率呈浓度及时间依赖关系;流式细胞术检测结果表明10、12.5mg.mL-1的贝伐单抗对细胞作用48h后,细胞凋亡明显,凋亡率分别为18.43±1.93%、26.98±2.21%,Western blot表明示贝伐单抗可显著下调pro-caspase-3蛋白,并呈浓度依赖关系。结论贝伐单抗可通过激活caspase-3而抑制HPF增殖、诱导其凋亡,呈剂量依赖性。

OBJECTIVE To observe the apoptosis and the expression of caspase-3 induced by Bevacizumab on Human Pterygium Fibmblasts（HPF） cultured in vitro. METHODS HPF were cultured in vitro. Bevacizumab was administered at a concentration ranging at 2. 5,5,7.5,10,12. 5mg·mL-1 for different time （24h ,48h and 72h）, and Fibroblast inhibition was assessed using the MTr colorimetrie assay. Flow cell cycle and Cell apoptosis were in- vestigated by flow-cytometry method. The expression of caspase-3 protein was detected by Western blot. RESULT Compared with the control group, the cell growth in experimental groups was inhibited significantly in the groups with concentration ranging at 7.5,10, 12. 5mg ·mL-1 （ P 〈 0. 01 ）, the inhibition ratios were 10. 49%, 17.03% and 26. 69% in respective groups when treated by Bevaeizumab for 48hs. The cell proliferation inhibition rate showed a dose-and time-dependent manner. Obvious apoptosis was observed by flow cytometry in 10 and 12. 5mg · mL-1 groups after 48ha,the apoptosis ratios were 18.43 ±1. 93% and 26. 98±2. 21% respectively. Western blot showed that the expression of pm-caspase-3 protein was decreased dose-dependertly. CONCLUSION Bevacizumab can in- hibit the proliferation of PHF,induee cell apoptosis,which may relate with the increasing of caspase-3 activity.