α_5、β_1整合素亚基在肺腺癌细胞凋亡中作用的实验研究

Experimental Study of the Function of α_5、β_1 Integrin Subunits on Apoptosis of Pulmonary Adenomatosis Cells

目的 探讨α5、β1整合素亚基对体外培养的肺癌细胞凋亡的作用及相关关系。方法 实验组以特异的抗α5、β1整合素亚基单克隆抗体 (McAb - 195 6Z、Sc- 9970 )分别阻断体外培养的人肺腺癌细胞 (GLC - 82 )膜上α5、β1整合素亚基的功能 ,采用吖啶橙 (acridineorange ,AO)做荧光染色和TUNEL法检测细胞凋亡的情况 ,并计算出凋亡指数 (apoptosisindex ,AI)。 结果 人肺癌细胞GLC - 82培养 2 4、 4 8h的AI值分别为 (1 3±0 6 7) %和 (2 4± 0 84 ) %。用McAb - 195 6Z阻断α5整合素亚基的实验组 (浓度分别为 1 0 μg/ml、 5 0 μg/ml和10 0 μg/ml) 2 4hAI值分别为 (2 0± 0 6 6 ) %、(2 7± 0 6 7) %和 (2 8± 0 6 3) % ;4 8hAI值分别为 (3 4±0 96 ) %、 (4 7± 1 33) %和 (5 1± 1 19) % ;以Sc - 9970阻断 β1整合素亚基的实验组 (浓度分别为 2 0 μg/ml、4 0 μg/ml和 10 0 μg/m) ,2 4hAI值分别为 (2 6± 0 6 9) %、 (3 8± 1 0 3) %、和 (5 1± 0 99) % ;4 8hAI值分别为 (4 8± 0 91) %、 (7 0± 0 81) %、和 (9 4± 1 17) %。与对照组相比差异有显著性意义 (P <0 0 1)。结论 α5、β1整合素亚基可抑制肺腺癌GLC - 82细胞的凋亡 ,表现为AI值增加及一种量。

Objective To study the role and relationship of α 5?β 1-integrin subunit on the pulmonary adenomatosis cellular apoptosis of external culture.Methods When the α 5?β 1-integrin subunit function of GLC-82 cells was blocked respectively by specific monoclonal antibodies in the experiment groups,the cellular apoptosis was determinated by AO fluorescent staining and TUNEL method.Results The GLC-82 cell apoptosis index(AI) was(1.3±0.67)% and(2.4±0.84)% respectively after 24h and 48h. The AI was(2.0±0.66)%,(2.7±0.67)% and (2.8±0.63)% respectively after 24h in the α5-integrin subunit experimental group blocked by McAb-1956Z and the AI was(3.4±0.96)%,(4.7±1.33)% and(5.1±1.19)% after 48h. In the group blocked Sc-9970 (the concentration of anti-α 5-integrin subunit antibodies was 1.0μg/ml?5.0μg/ml and 10.0μg/ml respectively ), the AI was(2.6±0.69)%,(3.8±1.03)% and (5.1 ± 0.99 ) respectively after 24h and (4.8±0.91)%,(7.0±0.81)% and (8.4±1.17)% after 48h. Comparing with control group the difference was significantly(P<0.01).Conclusions α 5,β 1-integrin subunit can inhibit GLC-82 cellular apoptosis, manifested as the increased AI and dependent relation on the dosage and effect between the GLC-82 apoptosis and α 5,β 1-integrin subunits.