神经生长因子对腺样囊性癌细胞增殖的影响

目的:检测神经生长因子(nerve growth factor,NGF)在体外培养腺样囊性癌(adenoid cystic carcinoma,ACC)细胞中的表达,探讨NGF对ACC细胞增殖的作用。方法:运用蛋白质印迹分析技术(Western blotting)检测体外培养增殖期ACC细胞中NGF的表达。体外培养ACC细胞分为9组:对照组(细胞浓度分别为2×104个/mL、1×104个/mL、5×103个/mL、2.5×103个/mL,为第1～4组)、实验组(细胞浓度为2×104/mL,加入NGF抗体的浓度分别为100μg/mL、20μg/mL、4μg/mL和0.8μg/mL,为第5～8组)、空白对照组(纯培养液,为第9组),采用MTT法检测孵育72 h后,计数ACC细胞增殖的数量;方差分析比较各组细胞增殖数量的差异。结果:ACC细胞中表达NGF;100μg/mL、20μg/mL和4μg/mL浓度的NGF抗体可明显抑制ACC细胞增殖的数量,P<0.01;随着NGF抗体浓度的递减,代表细胞增殖数量的OD值逐渐增大;当NGF抗体的浓度减至0.8μg/mL时,与对照组相比,ACC细胞增殖数量有所减少,但差别不显著,P>0.05;4种NGF抗体浓度的实验组之间细胞增殖的差异不显著,P=0.187。结论:NGF参与ACC细胞的增殖;NGF抗体通过中和NGF以剂量依赖的方式抑制ACC细胞增殖,反证了NGF能以剂量依赖的方式促进ACC增殖。这可能是NGF促进ACC神经侵袭的机制之一。

Effect of Nerve Growth Factor on Adenoid Cystic Carcinoma Cell Proliferation

Objective: To detect the expression of nerve growth factor(NGF) in adenoid cystic carcinoma(ACC) cells in vitro,and its role in ACC cell proliferation.Methods: Expression of NGF was detected by Western blotting in multiplication period cells of ACC in vitro.ACC Cells cultured in vitro were divided into nine groups,four experiment groups at cell concentration of 2×104/mL were treated with four different NGF antibody concentration(100 μg/mL?20 μg/mL?4 μg/mL or 0.8 μg/mL) including four control groups at four cell concentrations(2×104/mL?1×104/mL?5×103/mL and 2.5×103/mL),and one blank control group.Amount of cell proliferation was measured by MTT,and the difference among groups was analyzed by variance analysis.Results: NGF were expressed in ACC Cells.In vitro,anti-NGF antibody at concentrations of 100 μg/mL 20 μg/mL and 4 μg/mL markedly suppressed ACC proliferation(P<0.01).As the NGF antibody concentrations decreased,the amount of ACC increased concomittantly.At the concentration of 0.8 μg/mL,NGF antibody significantly reduced the cells number of ACC(P>0.05).There were no significant difference of ACC proliferation among four experiment groups(P=0.187).Conclusion: Anti-NGF antibody showed supressive effect on ACC proliferation in vitro in a dose-dependent manner.This means,NGF may promote ACC proliferation in a dose-dependent manner.