有机性挥发物在肺癌组织和癌细胞株中的检测和分析

Determination of volatile organic compounds in lung cancer cell lines and lung cancer tissue

目的:通过检测肺癌组织与肺癌细胞株代谢过程中产生的挥发性有机化合物(volatileorganic compounds,VOCs),寻找肺癌特征性的气体标记物,为呼吸气体检测在肺癌早期诊断中的应用提供实验依据。方法:利用固相微萃取联合气相色谱质谱联用技术(solid-phase microextractionand gas chromatography-mass spectroscopy,SPME-GCMS)检测18例原发性肺癌患者的手术标本及A549、NCI-H446、SK-MES-1、BEAS-2B 4种细胞株培养液顶空气体中的VOCs,从中筛选与肺癌相关的特征性VOCs组分。结果:2-十五烷酮和十九烷在肺癌细胞株A549、NCI-H446、SK-MES-1中均有表达,2-十五烷酮的表达量分别为(1.382±0.171)×10-5mg/L、(1.681±0.190)×10-4mg/L和(2.835±0.401)×10-6mg/L;十九烷的表达量分别为(8.382±0.606)×10-6mg/L、(1.845±0.130)×10-5mg/L和(6.220±0.362)×10-6mg/L;二十烷在肺癌细胞株A549和NCI-H446中有表达,表达量分别为(8.313±1.130)×10-6mg/L、(1.020±0.141)×10-5mg/L;而此3种VOCs在对照组BEAS-2B中无表达。在18例肺癌组织中,癸烷、2-十五烷酮、十九烷、二十烷等12种VOCs高表达,其中2-十五烷酮的表达量为5.421×10-6mg/L^3.621×10-5mg/L,十九烷为5.805×10-6mg/L^1.830×10-5mg/L,二十烷为2.730×10-6mg/L^2.343×10-5mg/L;其表达水平与肿瘤分化程度无关(P>0.05)。二十烷在非鳞癌中的表达高于鳞癌,此结果与肺癌细胞株的表达情况相符。结论:2-十五烷酮、十九烷、二十烷、癸烷等12种VOCs在肺癌组织中高表达,其中2-十五烷酮、十九烷、二十烷与肺癌细胞株结果相符,提示上述物质可能为肺癌潜在的特征性标志物。

Objective： To identify the volatile organic compounds（VOCs） in lung cancer tissue and lung cancer cell lines.Methods： The lung cancer tissue samples from 18 patients were cultured and 4 lung cell lines（A549,NCI-H446,SK-MES-1,BEAS-2B） were also included in the study.Air samples in the headspace of culture flasks were analyzed for VOCs with solid-phase micro-extraction and gas chromatography-mass spectroscopy technique（SPME-GC/MS）.Results： Two kinds of VOCs 2-pen-tadecanone and nonadecane were detected in lung cancer cell lines A549,NCI-H446 and SK-MES-1.The concentration of 2-pentadecanone were（1.382±0.171）×10^-5 mg/L,（1.681±0.190）×10^-4 mg/L and（2.835±0.401）×10^-6 mg/L,respectively;the concentrations of nonadecane were（8.382±0.606）×10^-6 mg/L,（1.845±0.130）×10^-5 mg/L and（6.220±0.362）×10-6 mg/L）,respectively.The eicosane was detected in A549 and NCI-H446 with the concentration of（8.313±1.130）×10^-6 mg/L and（1.020±0.141）×10^-5 mg/L）,respectively.All the 3 VOCs were not detected in cell line BEAS-2B.The concentrations of 12 VOCs including decane,2-pentadecanone,nonadecane and eicosane were high in 18 lung cancer tissue samples;the concentrations of 2-pentadecanone were 5.421×10^-6 mg/L-3.621×10^-5 mg/L,those of nonadecane were 5.805×10^-6 mg/L-1.830×10^-5 mg/L,those of eicosane were 2.730×10^-6 mg/L-2.343×10^-5 mg/L.There were no differences of VOCs levels among patients with different cancer differentiation（P〉0.05）.The concentration of eicosane in the non-squamous carcinoma was higher than that in squamous carcinoma,the same results were confirmed in the lung cancer cell lines.Conclusion： This study has identified VOCs produced by lung cancer tissue,which may support to use breath test as a complementary noninvasive diagnostic method for lung cancer.