肺癌患者肺泡巨噬细胞一氧化氮合酶mRNA表达及其抗肿瘤活性研究

EXPRESSION OF NITRIC OXIDE SYNTHASE mRNA AND ANTITUMOR ACTIVITY OF ALVEOLAR MACROPHAGES FROM LUNG CANCER PATIENTS

目的了解一氧化氮（ＮＯ）在肺癌患者肺泡巨噬细胞（ＡＭ）抗肿瘤功能的作用以及ＡＭＮＯ活性。方法通过支气管肺泡灌洗（ＢＡＬ）获取人ＡＭ；ＭＴＴ法观察一氧化氮合酶（ｉＮＯＳ）抑制剂Ｌ┐单甲基精氨酸（ＬＮＭＭＡ）对ＡＭ细胞毒功能的影响；镀铜镉还原——Ｇｒｉｅｓｓ法检测ＡＭ培养上清液、支气管肺泡灌洗液（ＢＡＬＦ）中ＮＯ含量；ＲＴ┐ＰＣＲ技术测定ＡＭｉＮＯＳｍＲＮＡ表达。结果（１）在ＬＮＭＭＡ存在条件下，肺癌患者ＡＭ细胞毒作用明显减弱（４３％ｖｓ２１％，Ｐ＜０．００１）。（２）肺癌患者荷瘤侧肺ＢＡＬＦ及ＡＭ培养上清液中ＮＯ含量均明显低于非荷瘤侧肺（Ｐ＜０．０５，Ｐ＜０．０１），更低于对照组（Ｐ＜０．００１，Ｐ＜０．０１）。（３）ＡＭ表达ｉＮＯＳｍＲＮＡ，肺癌组（４１％）明显低于对照组（７８％，Ｐ＜０．０５）。（４）经粒一巨细胞集落刺激因子（ＧＭ┐ＣＳＦ）刺激后肺癌患者ＡＭ细胞毒作用增强，ｉＮＯＡｍＲＮＡ表达增加（４１％ｖｓ６８％，Ｐ＜０．０１）且ＮＯ生成增多（６３ｎｍｏｌ／Ｌｖｓ８５ｎｍｏｌ／Ｌ，Ｐ＜０．００１），但后者仍低于对照组（Ｐ＜０．０５，Ｐ＜０．０１）。结论ＮＯ在肺癌患者ＡＭ介导的抗肿瘤效应中起着重要作用；肺?

Objective:The role of nitric oxide(NO)as an effector of alveolar macrophages(AMs) mediated tumoricidal activity and the mRNA expression of inducible nitric oxide synthase (iNOS) on AMs were investigated.Methods:AMs were obtained from 27 patients with primary lung cancer and 18 control cases with nonmalignant pulmonary diseases by bronchoalveolar lavage (BAL). The tumoricidal effect of AM was compared by using MTT in the presence or absence of N G monomethyl L arginine(LNMMA).Nitrite and nitrate(NO - 2/NO - 3)in original BAL fluid (BALF) and cell free supernatant were measured. AMs were analyzed for mRNA expression of iNOS by reverse transcription polymerase chain reaction (RT PCR).Results:On the presence of LNMMA the cytotoxicity exhibited by AMs significantly decreased( P <0 001).The level of NO - 2/NO - 3 was lower in both BALF and supernatants from the tumor bearing lung than from nontumor bearing lung P <0 01, P <0 05 ,and was much lower than control P <0 001, P <0 01 The iNOS mRNA expressed in AMs from 9/22(41%) patients with lung cancer and 14/18(78%) controls( P <0 005).After treating with GM CSF,lung cancer AMs demonstrated enhanced cytotoxicity ( P <0 001) and increased expression rates in iNOS mRNA.The level of NO - 2/NO - 3 in cell free supernatants was significantly increased ( P <0 001).However the expression rates and levels of NO - 2/NO - 3 were still lower in lung cancer patients than in control.Conclusion:NO plays an important role in AM mediated tumor cell cytotoxicity.There may be existed some defective AMs in tumor region. The expression of iNOS mRNA of AMs can be increased and cytotoxicity of AMs can be enhanced in vitro by GM CSF stimulation.