摘要
目的 建立白介素 8(IL 8)基因表达的绝对定量实时PCR检测技术。方法 应用实时定量PCR和ELISA方法检测 2株人肺癌细胞H4 6 0及A5 4 9中IL 8mRNA表达及蛋白分泌。同时对 9例正常肺及 4 4例非小细胞肺癌 (NSCLC)组织中IL 8mRNA进行分析。结果 应用实时定量PCR方法能够对肺癌细胞中IL 8mRNA拷贝数进行定量分析 ,IL 8表达值与蛋白分泌量呈正相关。正常肺组织中IL 8基因表达值为 4 .87± 1.6 9(单位 :拷贝数 / 10 4GAPDH拷贝 ) ,肺癌组织为 17.0 4± 2 3.96 ,两组间差异有统计学显著意义 (P =0 .0 0 2 )。若以正常肺组织均数的 2倍 (9.74 )和 4倍 (19.4 8)分别作为IL 8阳性表达及过表达的界限 ,5 2 .3% (2 3/ 4 4例 )的肺癌组织IL 8表达增高 ,其中IL 8阳性表达率为 2 9.5 % (13/ 4 4例 ) ,过表达率为 2 2 .7%(10 / 4 4例 )。统计分析显示 ,在女性、鳞癌和晚期肿瘤 (III期及T2 ～T4)中IL 8过表达率明显增高 ,具有统计学显著差异 (P <0 .0 5 )。结论 应用实时定量PCR技术能够对IL 8表达水平进行定量分析 ,IL 8过表达与患者性别、组织类型和病期密切相关。
Objective To develop a method for absolute quantification of interleukin 8 (IL-8) mRNA by using of real-time polymerase chain reaction (PCR). Methods The IL-8 mRNA and protein expression in 2 human lung cancer cell lines H460 and A549 were evaluated by real-time PCR and ELISA, respectively. The IL-8 mRNA expression in 9 cases of normal lung tissue and 44 cases of non-small cell lung cancer (NSCLC) were examined. Results The IL-8 mRNA copy number in a given sample can be measured by real-time PCR. The gene expression of IL-8 is correlated with its protein secretion. The normalized value of IL-8 expression was 4.87±1.69 (copies/10 4 GAPDH copies) in normal lung tissue and 17.04±23.96 in NSCLC, respectively. The difference between these two groups is statistically significant (P = 0.002). Using 9.74 and 19.48 as cut-off points for positive expression and overexpression of IL-8, 52.3% (23/44 cases) of NSCLC were found to express an increased level of IL-8, among which 29.5% (13/44 cases) were defined as positive expression and 22.7% (10/44 cases) as overexpression. Statistical analysis indicated that IL-8 overexpression was significantly increased in females, squamous histology, and late stages of disease ( P<0.05). Conclusion The IL-8 gene expression can be determined by real-time PCR technique. IL-8 overexpression is correlated with gender, histopathology and stages of the disease.
出处
《肿瘤》
CAS
CSCD
北大核心
2004年第1期3-5,共3页
Tumor
基金
上海市医学发展基金重大项目资助 (编号 :99ZDII0 0 1)
