吲哚胺2，3-双加氧酶对HepG2细胞生物学行为影响的研究
发布时间：2019-07-02 13:48
【摘要】：目的:构建IDO稳定表达细胞,应用体外细胞培养的方法,研究吲哚胺2 ,3-双加氧酶对肝癌细胞的生物学特点的影响。 方法:实验分为转染pcDNA3.1IDO的pcDNA3.1HepG2细胞组、转染空载体的pcDNA3.1HepG2细胞组和HepG2细胞组,各组细胞体外培养,观察细胞的形态学变化;采用流式细胞术检测细胞周期并计算细胞增殖指数;用细胞黏附试验和transwell小室侵袭试验分别检测各组细胞的增殖能力、细胞与细胞外基质黏附性的改变以及侵袭能力的变化。 结果:pcDNA3.1IDO-HepG2细胞与HepG2细胞和pcDNA3.1HepG2细胞相比,形态变为长梭形,伪足明显增多延长,前二者形态相似。流式细胞生长周期实验表明,HepG2、pcDNA3.1 HepG2细胞与pcDNA3.1-IDO HepG2相比,后者细胞增殖指数上升,细胞增殖能力差别有统计学意义(P 0.01)。细胞黏附试验表明,HepG2和pcDNA3.1 HepG2细胞的黏附性分别为0.93±0.18,1.07±0.14,较pcDNA3.1-IDO HepG(1.18±0.36)明显减小,差别有统计学意义(P 0.01)。侵袭试验表明,pcDNA3.1-IDO HepG2细胞侵袭能力增强,破坏matrigel基质蛋白胶层进入下室内的细胞数明显增多,HepG2、pcDNA3.1 HepG2和pcDNA3.1-IDO HepG2细胞分别为185. 60±15.58、198.80±12.60、269.00±21.60,前二者与后者比较差异具有统计学意义( P 0. 01)。 结论:IDO改变了HepG2细胞的生物学特点,明显促进HepG2细胞增殖能力、黏附能力和体外侵袭能力。
[Abstract]:Aim: to construct IDO stably expressed cells and to study the effect of indoleamine 2,3-dioxygenase on the biological characteristics of HCC cells by cell culture in vitro. Methods: the cells were divided into pcDNA3.1IDO pcDNA3.1HepG2 cell group, empty vector pcDNA3.1HepG2 cell group and HepG2 cell group. The cells in each group were cultured in vitro to observe the morphological changes of the cells, and the cell cycle was detected by flow cytometry and the cell proliferation index was calculated. Cell adhesion test and transwell chamber invasion test were used to detect the proliferation, adhesion and invasion of cells to extracellular matrix. Results: compared with HepG2 cells and pcDNA3.1HepG2 cells, the morphology of pcDNA3.1IDO-HepG2 cells became long fusiform, and the pseudopods were significantly increased and prolonged, and the morphology of the former two cells was similar. Flow cytometry showed that the proliferation index of HepG2,pcDNA3.1 HepG2 cells was higher than that of pcDNA3.1-IDO HepG2 cells, and the difference of cell proliferation ability was statistically significant (P 0.01). Cell adhesion test showed that the adhesion of HepG2 and pcDNA3.1 HepG2 cells was 0.93 卤0.18 and 1.07 卤0.14, respectively, which was significantly lower than that of pcDNA3.1-IDO HepG (1.18 卤0.36). The invasion test showed that the invasiveness of pcDNA3.1-IDO HepG2 cells was enhanced, and the number of cells that destroyed the matrix protein layer of matrigel into the lower chamber was significantly increased, and the number of HepG2,pcDNA3.1 HepG2 and pcDNA3.1-IDO HepG2 cells was 185, respectively. 60 卤15.58198.80 卤12.60269.00 卤21.60. there was significant difference between the first two and the latter (P 0. 0). 01) Conclusion: IDO can change the biological characteristics of HepG2 cells and promote the proliferation, adhesion and invasion of HepG2 cells in vitro.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R392
本文编号：2508980
[Abstract]:Aim: to construct IDO stably expressed cells and to study the effect of indoleamine 2,3-dioxygenase on the biological characteristics of HCC cells by cell culture in vitro. Methods: the cells were divided into pcDNA3.1IDO pcDNA3.1HepG2 cell group, empty vector pcDNA3.1HepG2 cell group and HepG2 cell group. The cells in each group were cultured in vitro to observe the morphological changes of the cells, and the cell cycle was detected by flow cytometry and the cell proliferation index was calculated. Cell adhesion test and transwell chamber invasion test were used to detect the proliferation, adhesion and invasion of cells to extracellular matrix. Results: compared with HepG2 cells and pcDNA3.1HepG2 cells, the morphology of pcDNA3.1IDO-HepG2 cells became long fusiform, and the pseudopods were significantly increased and prolonged, and the morphology of the former two cells was similar. Flow cytometry showed that the proliferation index of HepG2,pcDNA3.1 HepG2 cells was higher than that of pcDNA3.1-IDO HepG2 cells, and the difference of cell proliferation ability was statistically significant (P 0.01). Cell adhesion test showed that the adhesion of HepG2 and pcDNA3.1 HepG2 cells was 0.93 卤0.18 and 1.07 卤0.14, respectively, which was significantly lower than that of pcDNA3.1-IDO HepG (1.18 卤0.36). The invasion test showed that the invasiveness of pcDNA3.1-IDO HepG2 cells was enhanced, and the number of cells that destroyed the matrix protein layer of matrigel into the lower chamber was significantly increased, and the number of HepG2,pcDNA3.1 HepG2 and pcDNA3.1-IDO HepG2 cells was 185, respectively. 60 卤15.58198.80 卤12.60269.00 卤21.60. there was significant difference between the first two and the latter (P 0. 0). 01) Conclusion: IDO can change the biological characteristics of HepG2 cells and promote the proliferation, adhesion and invasion of HepG2 cells in vitro.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R392
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