ABSTRACT
<p><b>OBJECTIVE</b>To observe autophagy induced by starvation in non-small cell lung cancer A459 and 95D cells.</p><p><b>METHODS</b>A549 and 95D cells in logarithmic growth in 1640 medium were cultured in Earle's balanced salt solution (EBSS) for 0, 1, 2, 3, 4 or 5 h. Autophagosome formation in the cell culture was observed by MDC fluorescent staining, and the expression of microtubule-associated protein 1 light chain 3 (LC3) and Beclin1 in the cells were detected using Western blotting.</p><p><b>RESULTS</b>Compared with the control cells, the cells with prolonged starvation showed increased MDC-positive cells and autophagosome formation. The expression of Beclin-1 and the LC3-II/LC3-I ratio also increased as the starvation prolonged, reaching the peak levels at 3 h and 4 h, respectively.</p><p><b>CONCLUSION</b>Autophagy can be induced by starvation in A549 and 95D cells in correlation with the expression of autophagy-related proteins LC3 and Beclin-1. These cell models of nutritional deficiency-induced autophagy may allow for a better understanding of the role of autophagy in the development of non-small cell lung cancer.</p>
Subject(s)
Humans , Apoptosis Regulatory Proteins , Metabolism , Autophagy , Beclin-1 , Carcinoma, Non-Small-Cell Lung , Pathology , Cell Line, Tumor , Membrane Proteins , Metabolism , Microtubule-Associated Proteins , MetabolismABSTRACT
Objective To purify and identif y Platanus acerifoli wild pollen. Methods We carried out intracutaneous test with Platanus pollen extract in 30 patients with allergic as thma who visited our hospital from March to May 2003. Seven subjects who had bee n diagnosed as having Platanus pollen-induced asthma were enrolled. Platanus po llen proteins were separated by gel filtration with Sephadex-G-100. To charact erize allergenic components, Platanus pollen extract was analyzed by means of so dium dodecylsulfate-polyacrylamide gel electrophoresis followed by immunoblotti ng. Results To purity the pollen we separated Platanus poll en extract in a first purification step by using gel filtration with Sephadex G -100. Two elution peaks were observed. Twelve percent SDS-PAGE analysis showed more than 10 protein bands whose molecular mass (Mr) ranged from 16 ku to 71 ku. Six bands abundant with protein at 71, 50, 35, 39, 22 and 16 ku were observed. On SDS-PAGE, the proteins of the first peak whose Mr we re 71, 50, 35, 39, and 22 ku and that of the second peak was 16 ku. SDS-PAGE and IgG-immunoblotting analysis with seven sera showed 4 IgG-binding component s whose Mr was 50, 39, 22 and 16 ku. The protein bands whose Mr was 50 ku and 22 ku had the highest binding capacity. Conclusion The strongest activity exists in the first peak which can be the major sensiti zing components and there is mild allergic activity in the second peak which is the minor sensitizing components.