碱性成纤维细胞生长因子诱导牛晶状体上皮细胞增殖细胞核抗原的表达及对晶状体上皮细胞增殖作用的研究

A study of the expression of PCNA and the proliferation of bovine lens epithelial cell induced by bFGF

目的 观察碱性成纤维细胞生长因子 (basicfibroblastgrowthfactor,bFGF)诱导牛晶状体上皮细胞 (bovinelensepithelialcell,BLEC)增殖细胞核抗原 (proliferatingcellnuclearantigen ,PCNA)表达及对BLEC增殖的作用。方法 取体外培养的第 2代BLEC ,加入不同浓度的bFGF(0 0 1～ 10 0 0 0μg/L) 共同培养 2 4～ 96h后 ,用3 H TdR掺入法测定BLECDNA合成率 ,用流式细胞仪检测细胞周期和PCNA的表达。结果 不同浓度的bFGF有促进BLEC增殖作用 ,并呈剂量时间依赖性 ,当浓度为10 0 0 μg/L的bFGF作用BLEC 2 4～ 4 8h后 ,晶状体上皮细胞的3 H TdR掺入值分别为 11772 5和10 988 2 ,较对照组的 6 5 5 0 5增加约一倍 ,两组比较差异有显著意义 (P <0 0 5 )。bFGF作用BLEC可使PCNA表达率上升 (77 4 0 % ) ,并使BLEC周期发生变化 ,S期细胞和G2 /M期细胞比例分别提高至2 3 4 1%和 2 0 76 % ,与对照组比较 ,差异有显著意义 (P <0 0 5 )。结论 bFGF通过上调BLEC的PCNA表达 ,改变细胞周期 。

Objective To observe the effect of the expression of proliferating cell nuclear antigen (PCNA) induced by basic fibroblast growth factor (bFGF) on the proliferation of bovine lens epithelial cells (BLECs) Methods The second passage BLECs were cultured with different concentrations of bFGF (0 01 100 00 μg/L)for 24 96 hours, the DNA synthesis of BLEC was measured by 3?H TdR incorporation, and the expression of PCNA and the cell cycle were counted by the flow cytometer Results bFGF promoted the proliferation of BLEC with concentration and time dependent manner The 3?H TdR incorporation of BLEC was increased to 11 772 5 and 10 988 2 respectively much higher than the control group (6 550 5). The addition of bFGF 10 00 μg/L for 24 48 hours up regulated the expression of PCNA (77 40%), and caused the BLEC population to increase in S phase (23 41%) and G 2/M phase (20 76%) Comparing with the control group, there was significant difference ( P <0 05) Conclusion bFGF stimulateds the proliferation of BLEC by up regulating the expression of PCNA and changing the cell cycle