ABSTRACT
OBJECTIVE@#To investigate the effect of basic fibroblast growth factor (FGF-2)on survivin and subcellular location of Smac in human small cell lung cancer (SCLC) cell NCI-H446.@*METHODS@#Western blot was used to detect the expression of survivin protein induced by FGF-2. The release of Smac from mitochondria to cytoplasm affected by FGF-2 was observed by Western blot and immunofluorescence. Apoptosis of NCI-H446 cells was detected with flow cytometry and Hoechst 33258 staining.@*RESULTS@#The expression of survivin could be up-regulated in response to FGF-2 treatment in NCI-H446 cells, and the level of survivin expression is related to the concentration and time of FGF-2 treatment. FGF-2 could inhibit the release of Smac from the mitochondria to cytoplasm induced by serum starving. FGF-2 could inhibit the apoptosis induced by serum starving.@*CONCLUSION@#FGF-2 up-regulates the expression of survivin protein in NCI-H446 cells, and blocks the release of Smac from mitochondria cytoplasm. Survivin and Smac might play important roles in the apoptosis inhibited by FGF-2.
Subject(s)
Humans , Apoptosis , Apoptosis Regulatory Proteins , Cytoplasm , Metabolism , Fibroblast Growth Factor 2 , Pharmacology , Inhibitor of Apoptosis Proteins , Intracellular Signaling Peptides and Proteins , Metabolism , Lung Neoplasms , Metabolism , Pathology , Microtubule-Associated Proteins , Mitochondria , Metabolism , Mitochondrial Proteins , Metabolism , Small Cell Lung Carcinoma , Metabolism , Pathology , Survivin , Tumor Cells, CulturedABSTRACT
OBJECTIVE@#To determine the expressions of survivin and proliferating cell nuclear antigen (PCNA)in non-small cell lung cancer (NSCLC) and to explore its clinical pathological significance.@*METHODS@#Immunohistochemical SP method was used to detect the expressions of survivin and PCNA in 43 patients with NSCLC and 15 normal epithelial tissues of the lung. PCNA labeling proliferative index was assessed. Forty-three patients with NSCLC were followed up for more than 5 years.@*RESULTS@#The positive expression of survivin in NSCLC (79.1%) was significantly higher than that in normal epithelial tissues of the lung (P 0.05). The mean proliferative index of PCNA in NSCLC was much higher than that in normal epithelial tissues of the lung (P < 0.01). A positive correlation was present between the proliferative index and the tumor size, lymph node metastase, and clinical stage (P <0.01), while a negative correlation between the proliferative index and survival time (P <0.01). There was no correlation between proliferative index and age, sex, site, histological type and grade. The proliferative index was larger in patients with moderate or strong positive survivin expression than that in patients with negative or weak survivin expression (P < 0.05).@*CONCLUSION@#Over expression of survivin and PCNA is closely correlated to the progression and prognosis of patients with NSCLC, which is helpful to evaluate the progression of cancer and to predict the prognosis of NSCLC. The up-regulation of survivin expression and its close relationship with the cell proliferation in NSCLC suggest that survivin may play an important role in the carcinogenesis and development of lung cancer.
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Biomarkers, Tumor , Carcinoma, Non-Small-Cell Lung , Metabolism , Inhibitor of Apoptosis Proteins , Lung Neoplasms , Metabolism , Microtubule-Associated Proteins , Genetics , Neoplasm Proteins , Genetics , Neoplasm Staging , Prognosis , Proliferating Cell Nuclear Antigen , Genetics , SurvivinABSTRACT
OBJECTIVE@#To determine the effects of exogenous RASSF1A gene on the proliferation and expression of P65 and subunit of NF-kappaB, in lung adenocarcinoma cell line A549.@*METHODS@#pcDNA3.0-RASSF1A and pcDNA3. 0 were introduced into A549 cell line by lipofectin transfection, and the A549 cells stably expressing RASSF1A gene were established by G418 selection. The expression of RASSF1A was detected by Western blotting. The cytobiologic characterizations of the positive clone were analyzed by methythiazoletertraolium (MTT) assay and cytometry. The expressing of P65 was analyzed by RT-PCR and Western blotting.@*RESULTS@#A549 cells stably expressing RASSF1A protein were established by lipofection mediated transfection and selected for further study. Compared with the nontransfected and vector transfected cells, the positive clone cells grew more slowly. Flow cytometric data showed that more positive clone cells went into phase G0/G1 and fewer cells went into phase S. The expression of P65 in nuclear protein in positive clone cells was lower than that of the control group while there was no obvious difference between the expression of p65 mRNA and P65 protein in total protein among the 3 groups.@*CONCLUSION@#RASSF1A gene might suppress the proliferation of A549 cells through blocking the activity of P65 protein.