Avastin对翼状胬肉成纤维细胞增殖的影响  

Effects of Avastin on human pterygium fibroblasts proliferation

作　　者：刘祎[1] 沙翔垠[1] 彭娟[1] 文晔[1] 宋莉[1] 

机构地区：[1]广州医科大学附属第二医院眼科教研室,广东省广州市510260

出　　处：《眼科新进展》2014年第9期813-816,共4页Recent Advances in Ophthalmology

基　　金：广东省科技计划项目基金资助(编号:20100309)~~

摘　　要：目的评价Avastin(贝伐单抗)在体外对翼状胬肉成纤维细胞的增殖抑制作用以及细胞毒性。方法通过不同浓度Avastin(0.25g·L-1、0.50g·L-1、1.00g·L-1、1.50g·L-1、2.50g·L-1)处理人翼状胬肉成纤维细胞12h、24h、48h、72h,观察成纤维细胞的增殖抑制、药物细胞毒性以及凋亡蛋白的表达。WST-1试剂检测细胞增殖,LDH试剂盒检测细胞毒性,Western blot方法检测增殖蛋白以及凋亡蛋白的表达。结果 48h及72hAvastin组与正常组相比光密度(OD)值差异有统计学意义(P=0.034、P=0.016)。与12h相比,0.25g·L-1 Avastin组及0.50g·L-1 Avastin组作用24h、48h、72h后细胞增殖抑制率均无明显提高(均为P>0.05),1.50g·L-1 Avastin组及2.50g·L-1 Avastin组作用48h、72h后细胞增殖抑制率均明显提高(均为P<0.05)。与0.25g·L-1 Avastin组相比,0.50g·L-1 Avastin组在各个时间点(除12h外)细胞增殖抑制率均无明显提高(均为P>0.05),1.50g·L-1 Avastin组、2.50g·L-1 Avastin组在各个时间点抑制率均明显提高(均为P<0.05)。Avastin各浓度组与LDH最大释放组(加入裂解酶)对翼状胬肉成纤维细胞LDH释放率差异均有统计学意义(均为P<0.05),各浓度组之间差异均无统计学意义(均为P>0.05)。Avastin可抑制p-CyclinD1表达,促进Caspase3的表达,与细胞对照组相比差异均具有统计学意义(均为P<0.05)。结论 Avastin可以通过抑制p-CyclinD1以及促进Caspase3的表达抑制翼状胬肉成纤维细胞的增殖,但无细胞毒性,进一步证实了该药物的安全有效性。Objective To evaluate the anti-proliferation and cytotoxicity of Bev- acizumab （Avastin） on human pterygium fibroblasts （HPF）. Methods HPF were treated with Avastin （0.25 g · L^- 1, 0.50 g · L^- 1,1.00 g·L ^-1 , 1.50 g · L^-1, 2.50 g · L^-1 ） for 12 hours,24 hours,48 hours,72 hours. Cell proliferation was measured with the water soluble tetrazolium salt （ WST-1 ） assay. Cell cytotoxicity assay was measured by LDH release assay. Expression of p-CyclinDl and Caspase3 were detected by Western blot. Results Compared with normal control, there were statistical differences in OD value at 48 hours and 72 hours of Avastin group （P =0.034,0. 016）. Compared with treat- ment for 12 hours,no significant difference was found for the inhibitory effect on proliferation in the groups for 24 hours,48 hours and 72 hours of 0. 25 g · L^-1 and 0. 50 g · L^-1 Avastin （ all P 〉 0.05 ）. However,the inhibitory effect on proliferation was significantly increased when treated with high concentrations （ 1.50 g · L^- 1, 2.50 g · L^- 1 ） at 48 hours and 72 hours （all P 〈0.05 ）. Compared with 0.25 g · L^-1 Avastin,the difference of inhibitory effect on proliferation of 0.50 g · L^- 1 Avastin was not significant at each time （ except for 12 hours） （all P 〉0. 05） ,but there were significant increase in high concentration groups （ 1.50 g · L^- 1,2.50 g · L^- 1 ） （ all P 〈 0.05 ）. Compared with the max LDH release group ,there were statistical differences in LDH release rate of each Avastin concen- tration group （all P 〈 0.05） ,but there was no statistical difference between each concentration groups（ all P 〉 0.05 ）. Avastin could significantly inhibit the expression of p-Cy- clinD1, enhanced the expression of Caspase 3,there were statistical differences compared to the control group （ all P 〈 0.05 ）. Conelusion Avastin can inhibit the proliferation of HPF through decreasing the expression of p-CyclinD1 and enhancing the expression of Caspase 3 with no cytotoxi

关 键 词：贝伐单抗 翼状胬肉 细胞增殖 细胞毒性 CYCLIND1 CASPASE 3 

分 类 号：R733.7[医药卫生—肿瘤]