乙酰胆碱联合长春新碱对人类肺腺癌细胞株A549生长及凋亡的影响

Effects of acetylcholine combined with vincristine on the proliferation and apoptosis of A549 cells

目的探讨乙酰胆碱联合长春新碱（VCR）对肺腺癌细胞生长及化疗的影响，检测其是否通过影响bel-2及p53发挥作用。方法将乙酰胆碱单独或与VCR（浓度为IC650）联合作用于肺腺癌细胞株A549，测定其对A549细胞的抑制率。用免疫组织化学法及Westernblot法测定乙酰胆碱联合VCR对bcl-2及p53蛋白表达的影响。结果0．1、1、10μmol／L的乙酰胆碱对A549细胞的抑制率分别为（0．050±0．032）％、（0．101±0．021）％、（0．169±0．015）％；0．1、1、10μmol／L的乙酰胆碱分别与0．2μg／mlVCR联合，对肿瘤细胞的抑制率分别为（0．529±0．023）％、（0．545±0．011）％、（0．589±0．015）％，与VCR组比较差异均有统计学意义（q’值分别为1．09、1．37、1．83，均P〈0．05），0．1。10μmol／L的乙酰胆碱单独作用于A549细胞表现出浓度依赖性的抑制作用，与VCR联合后显著提高A549对VCR的敏感性（P〈0．05）；此外，联合用药后各组bcl-2及p53蛋白表达均降低（P〈0．05）。结论乙酰胆碱单独及联合VCR均能抑制A549细胞的生长，其机制可能是乙酰胆碱与VCR协同或相加作用影响了bel-2及p53蛋白的表达。

Objective To study the effects of acetylcholine combined with vincristine （VCR） on A549 cells, furthermore to study the influence that is due to affecting the expression of the two proteins bcl-2 and p53. Methods MTT assay was used to analyze growth inhibition effect of aeetylcholine alone and combined with IC50 VCR on A549 cells. The expression of bel-2 and p53 was measured by immunohistological chemistry teehnique（IHC） and Western blot. Results The inhibition rates for A549 cells with 0.1, 1 and 10μmol/L acetylcholine were （0.050±0.032）%, （0.101±0.021）%, （0.169±0.015）%. The inhibition rates for tumor cells with 0.1, 1 and 10 μmol/L acetylcholine joint 0.2 μg/ml VCR were （0.529±0.023）%, （0.545±0.011）%, （0.589± 0.015）%,the difference was statistically significant compared with the VCR group （q＇values were 1.09,1.37, 1.83,P〈0.05）. Acetylcholine alone exerted inhibitory effect on A549 cells in a concentration dependent manner and significanrtly enhanced its sensitivity to VCR （P〈0.05）. acetylcholibne （0.1-10 p.mol/L）combined with IC50VCR decrased the expression of bel-2 and p53 （P〈0.05）. Conclusion Acetyleholine alone and combined with VCR can inhibit the growth of A549 cells. Significant synergistic effect between acetyleholine and VCR is found in inducing cell apoptosis by changing the expression of bel-2 and p53.