The Relationship Of Thymosin Beta With Metastasis And Prognosis Of Lung Cancer

INTRODUCTIONThe most cause of sufferers death is development to metastasis phase to tumor. Tumor cell movement and angiogenesis are essencial to realize invasion and metastasis of tumor cell. In the present study, thymosin beta can not only stimulate cell movement,but also influence angiogenesis and resist apoptosis. It can acculate invasion and metastasis straightly or indirectly by many ways. Therefore, it's becoming an hotspot of tumor metastasis mechanism and counter-measure.Expression of thymosin beta isnt much in normal tissues, while its expression in tumor clearly higher than expression of original tissue. But this expression altering universalism and early diagnosis and prognostic evaluation are still needed to research. At present, investigation is very few about theirs expression in tumors and theirs relationship . So we assessed the presence of thymosin beta in lung squama and adenocarcinoma and relationship them with clinic pathology factors and their roles in the invasion, metastasis and prognosis of lung cancer.MATERIALS AND METHODS1. Tissue samplesA total of seventy - six cases lung cancer with neighboring thirty cases ofnoncancerous tissue samples were obtained from patients underwent surgical resection in the Anshan Tumor Hospital between 1980 and 2001, fixed in 4% paraformaldehyde, embedded in paraffin. Twenty - five freshly taken lung cancer tissue samples with corresponding non - cancerous tissues were obtained from the First Affiliated hospital of China Medical University between 2003 - 2004.2. Clinic and patholic dataIn 76 cases, I II III IV stage were 18 16 35 7 cases respectively ;squa-ma cancer was 41 cases and adenocarcinoma was 35 cases; high differentiation was 16 cases, middle differentiation was 28 cases and low differentiation was 32 cases; lymophnode metastasis was 50 cases; blood metastasis was 45 cases ( brain metastasis:23 cases;bone metastasis: 10 cases derenal gland metastasis: 12 cases ) ; average survival time was 22. 84 months. In 25 fresh tissues, squma cancer was 9 cases and adenocarcinoma was 16 cases;high differentiation was 5 cases, middle differentiation was 13 cases and low differentiation was 7 cases.3. ReagentsThe primary antibody anti - human thymosin beta IgY antibody were provided by E. Lvaniov in Greece Immunopeptide Chemistry Lab" NCSR DEMOKRI-TOS". The HRP marked rabbit anti - chicken second antibody bought from Shanghai Casarray Company. The Ultro - sensitive S - P kit and DAB agent kit were bought from Fujian Maxin Biological Company. The probe of thymosin beta 15 mRNA and reagent kit in situ hybridization were bought from Wuhan Boston Biological Company.4. Methods4.1 Immunohistochemistry for Thymosin beta Immunohistochemical staining was performed using a Ultro - sensitive S - Pkit according to the instructions provided by the manufacturer. Color was developed using DAB/H2O solution. After the areas of tumor tissue had been chosen by random under low power, one hundred cells within tumors were counted in each of 10 fields at 200 magnification. Cases defined as positive were regarded by both staining intensity and the percentage of immunoreactive tumor cells.4.2 Situ hybridization of thymosin betal5 mRNAParaffin - embedded specimens (4μm) were deparaffinized and rehydra-ted. The steps of hybridization were performed using a kit according to the recommendations of the manufacturer.5. Statistical analysisStatistical Product and Services Solutions (SPSS) statistical software (version 10.0) was applied for data analysis. Chi - square test was used to compare the clinicopathological characteristics with the genes expression. Kappa - value estimated correlation of expression of thymosin betal5 protein and mRNA Postoperative survival periods were computed by the Kaplan - Meier method and compared by the Log rank test. A P - value less than 0.05 was considered significant. The Cox stepwise regression analysis of which the level of significance was set at P≤0. 1 was used to evaluate the statistical strength of independent association between selected covariates and patient survival.RESULTS1. Expressions of Tβ₄ Tβ₁0 Tβ₁5 protein in lung cancer and non -cancer tissuesIn lung cancer, Staining in immunoreactive tumor cells were observed for cytoplasmic showing diffusion. Tβ₄ and Tβ₁0 were particularly expressed in nu-cleolus. There were also some levels of expression in macrophages vascular en-dothelioids, lymphocytes,fibroblasts. the high expression rates of them were 76. 3% 72.4% 63.4% respectively In non -cancer tissues, Tβ₄ was low expression in alveolus epithelium and none of bronchus epithelium. Tβ₁0 and Tβ₁5 were no expression in both alveolus and bronchus epithelium.2. The relationship high expressions of Tβ₄ T_β10 Tβ₁5 protein with clinic patholic factorsThere was a significant negative correlation between the expression of both their protein and differentiation of lung cancer. Expressions oftheir were positive associated with TNM stage and metastasis. However, none of the three proteins was correlated with the age, sex, and tumor type.3. The relationship high expressions of Tβ₄ Tβ₁0 Tβ₁5 protein with prognosis...