TOB1对人类肺癌细胞增殖、转移和放射敏感性的影响及机制的研究临床医学专业论文.docxVIP

PAGE PAGE IV TOB1 对人类肺癌细胞增殖、转移和放射敏感性的影响及机制的研究 英文摘要 Studies on the effcets and mechanisms of TOB1 on the proliferation, metastasis and radiosensitivity of lung cancer cells Abstract Aim: To investigate the effects of the transducer of erbB-2.1 (TOB1) on the proliferation, metastasis and radiosensitivity of human lung cancer cells in vitro. Methods: TOB1 expression in eight human lung cancer cell lines (95-D, A549, NCI-H1299, NCI-H1975, NCI-H661, NCI-H446, NCI-H1395 and Calu-3) and the normal human bronchial epithelial (HBE) cell line was determined by Western blot and RT-PCR analysis. TOB1-overexpressed cell lines 95-D/TOB1, NCI-H1975/TOB1 were obtained using lipofectamine-induced TOB1 recombinant plasmid transfection and selective G418 cell culture. TOB1 knockdown was performed using RNAi in A549 cells. MTT assay, flow cytometry, wound healing and transwell invasive analysis were used to determine the effects of TOB1 on lung cancer cells proliferation, migration and invasion. The activity of MMP2 and MMP9 was measured using gelatin zymography assay. As for radiosensitivity, cell cycle and DNA repair was assessed in regards to radiation by flow cytometry and Immunofluorescence analysis. Western blot and immunopre- cipitation analysis were utilized to determine the related mechanisms. Results: The expression of TOB1 in the 8 human lung cancer cell lines were significantly decreased corresponding to the nomal HBE cells. TOB1 overexpression inhibited the proliferation, migration and invasion of 95-D cells, and upregulated the gelatinase activity of MMP2 and MMP9. TOB1 overespression reduced cell clonogenic survival, abrogated radiation-induced G2/M arrest and enhanced ?-H2AX foci in NCI-H1975 cells after ionizing radiation. Importantly, TOB1 knockdown demonstrated opposite effects in A549 cells. Immunoprecipitation confirmed the protein-protein interaction between TOB1 and PTEN. Western-blot revealed TOB1 regulates the expression of cellular EGFR and the acti