GM-CSF对肺癌细胞凋亡及肺泡巨噬细胞抗肿瘤功能的影响

The study on the effect of GM－CSF on apoptosis in lung cancer cells and on antitumor activity of alveolar macrophage in human

目的：了解GM－CSF对肺癌细胞的杀伤作用以及时其凋亡和对AM抗肿瘤功能的影响。方法：MTT观察GM－CSf对人肺腺癌细胞株（A549）细胞的杀伤作用以及对肺癌患者AM杀伤A549细胞的影响。透射电镜、凝胶电泳及片段裂解率观察经CM－CSF作用的A549细胞凋亡情况．镀钢镉还原法、放免法、比色法分别检测AM培养上清中NO、TNFα、SOD活性，并用RT．PCR检测AMiNOSmRNA表达。结果：GM－CSF对A549细胞有一定杀伤作用，并呈浓度依赖性。A549细胞核扭曲断裂，凋亡小体形成，DNA凝胶电泳可见梯状条带。AM经GM－CSF刺激后对A549细胞的杀伤率由14.35%增加至4352%（P＜0．001），其培养上清波中NO、TNFα的水平显著升高，AMiNOSmRNA表达明显增强。结论：GM－CSF可诱导A549细胞凋亡，刺激肺癌患者AM抗肿瘤功能增强。

Purpose: The effect of GM－CSF on apoptosis of lung cancer cells and on the antiumor activity of AMs was investigated. Methods: me tumoricidal ability of GM－CSF and AMs stimulated stimulated by GM－CSF from patients with lung cancer for human lung adenocarcinoma cell line (A549) cells was examined by MTT method. The morphological changes Of A549 cells were observed by transmission electron microscope (TEM ). DNA agarose gel electroplloresis was adopted to investigate A549 cells death from induced by GM－CSF DNA fragmention of apoptotic A549 cells was quantitatively measured. In the AM－free supernatants, the activity of NO. TNF－α and SOD was examined by coppercoated cadmium reduction and Griess, radioimmunoassay, xanthine－oxidase assay. respectively. RT－PCR was applied ic detect mRNA expression of iNOS in AMs from lung cancer patients. Results: The various concelltlution of GM－CSF all exhibited tumor cell cytotoxicity. The effect has close correlation with concentratioll (r = 0. 7858. P < 0. 01 ). TEM obselvation revealed that the nucleus were fragmented into pieces and apoptolic bodies were developed. Thers was specific DNA ladder pattern upon agarose gel electrophoresis. When AMs stimulated by GM－CSF the cytotoxicity of AMs was significantly' enhanced (P < 0. 001 ), and the activity of NO and TNF α was increased and the degree of mRNA expression of iNOS in AMs was enhanced. Conclusion: GM－CSF may induce apoptosis of A549 cells and enhance the tumoricidal ability of AMs lung cancer patients.