Real-time PCR And Immunohistochemistry Detecting The Expression Of Survivin In NSCLC And The Clinical Significance

Objective: We attempted to establish a quantitative Real-time fluorescen- ce quantitative PCR method. We examined survivin protein and gene expression in non-small cell lung cancer and normal lung specimens,then analyzed the relationship between survivin expression and relevant clinical and pathologic data.Methods:Immunohistochemistry staining and Real-time fluorescence quantitative PCR were used for detecting the expression of survivin protein and survivin mRNA in 40 cases NSCLC and 20 cases normal lung specimens,then analyzed the relationship between survivin expression and the clinical pathological characteristics,pathological types. SPSS (13.0) statistical software was applied for data analysis. The correlation between the expression of survivin and the major clinical pathological characteristics of NSCLC were analyzed by Chi-square test or Fisher exact probability, T test or one-way ANVOA. Postoperative survival period analysis were computed by the Kaplan-Meier method and compared by the Log-tank test.Results: 1.Immunohistochemistry results: (1) The positive expression rate of survivin in NSCLC specimens was 67.5% ,it was significantly higher than that in normallung tissue 5%(P=O.000).(2) The positive expression rate of survivin in NSCLC:①male was 62.5% and female 75%(P=0.408),②the patients under the age of 60 was 65.4% and those over the age of 60 was71.4% ( P=0.972 ) ,③the pulmonary squamous carcinoma was 66.7%,the adenocarcinoma was 68.8% and the squamous adenocarcinoma was 66.7%(P=0.991),④high histological grades was 71.4%,middle histological grades was 50% and low histological grades was 69%(P=0.739),⑤tumor size≤3cm was 80% and tumor size>3cm was 63.3%(P=0.599),⑥lymphnode metastasis was 68% and no lymphnode metastasis was 67.7%(P=0.931),⑦TNM stagesⅠwas 75%,Ⅱwas 60% andⅢwas 66.7%(P=0.873),⑧recurrence≤6 month was 83.3% and recurrence>6 month was64.7%(P=0.643).(3) Median survival period of patients with survivin positive was 24 monthes ,but patients of negative expression was 28 monthes(P=0.285). 2. Real-time fluorescence quantitative PCR results:(1) The expression of survivin mRNA in NSCLC 1.639±1.372 was significantly higher than that in benign disease lung tissue 0.580±0.487 (P=0.000).(2) The expression of survivin in NSCLC:①male was 1.605±1.521 and female 1.691±1.156(P=0.848),②the patients under the age of 60 was 1.501±1.154 and those over the age of 60 was 1.897±1.725(P=0.390),③the pulmonary squamous carcinoma was 1.613±1.807,the adenocarcinoma was 1.829±1.132 and the squamous adenocarcinoma was 1.413±1.224(P=0.737),④high histological grades was 1.120±0.730,middle histological grades was 1.450±0.472 and low histological grades was 1.765±1.554(P=0.616),⑤tumor size≤3cm was 1.732±1.559 and tumor size>3cm was 1.609±1.333 ( P=0.809 ) ,⑥lymphnode metastasis was 1.671±1.517and in no lymphnode metastasis was 1.587±1.137(P=0.855),⑦TNM stagesⅠwas 1.609±1.214,Ⅱwas 1.747±1.584 andⅢwas 1.600±1.423(P=0.962),⑧in recurrence≤6 month was 2.056±1.948 and in recurrence>6 month was 1.566±1.270(P=0.427).(3) Median survival period of patients with survivin mRNA hihger expression was 22.5 monthes ,but patients of lower expression was 24 monthes(P=0.325).Conclusion: We had successfully established a Real-time fluorescence quantitative PCR method for detecting of survivin gene in non-small cell lung cancer(NSCLC) . The survivin gene was expressed significantly higher in non-small cell lung cancer specimens than in normal lung specimens. The abnormal expressed survivin may play an important role in the genesis and development of lung cancer.The prognostic value of survivin in NSCLC is unclear,further studies had to be done.