【摘要】：目的:構(gòu)建IDO穩(wěn)定表達(dá)細(xì)胞,應(yīng)用體外細(xì)胞培養(yǎng)的方法,研究吲哚胺2 ,3-雙加氧酶對(duì)肝癌細(xì)胞的生物學(xué)特點(diǎn)的影響。 方法:實(shí)驗(yàn)分為轉(zhuǎn)染pcDNA3.1IDO的pcDNA3.1HepG2細(xì)胞組、轉(zhuǎn)染空載體的pcDNA3.1HepG2細(xì)胞組和HepG2細(xì)胞組,各組細(xì)胞體外培養(yǎng),觀察細(xì)胞的形態(tài)學(xué)變化;采用流式細(xì)胞術(shù)檢測(cè)細(xì)胞周期并計(jì)算細(xì)胞增殖指數(shù);用細(xì)胞黏附試驗(yàn)和transwell小室侵襲試驗(yàn)分別檢測(cè)各組細(xì)胞的增殖能力、細(xì)胞與細(xì)胞外基質(zhì)黏附性的改變以及侵襲能力的變化。 結(jié)果:pcDNA3.1IDO-HepG2細(xì)胞與HepG2細(xì)胞和pcDNA3.1HepG2細(xì)胞相比,形態(tài)變?yōu)殚L(zhǎng)梭形,偽足明顯增多延長(zhǎng),前二者形態(tài)相似。流式細(xì)胞生長(zhǎng)周期實(shí)驗(yàn)表明,HepG2、pcDNA3.1 HepG2細(xì)胞與pcDNA3.1-IDO HepG2相比,后者細(xì)胞增殖指數(shù)上升,細(xì)胞增殖能力差別有統(tǒng)計(jì)學(xué)意義(P 0.01)。細(xì)胞黏附試驗(yàn)表明,HepG2和pcDNA3.1 HepG2細(xì)胞的黏附性分別為0.93±0.18,1.07±0.14,較pcDNA3.1-IDO HepG(1.18±0.36)明顯減小,差別有統(tǒng)計(jì)學(xué)意義(P 0.01)。侵襲試驗(yàn)表明,pcDNA3.1-IDO HepG2細(xì)胞侵襲能力增強(qiáng),破壞matrigel基質(zhì)蛋白膠層進(jìn)入下室內(nèi)的細(xì)胞數(shù)明顯增多,HepG2、pcDNA3.1 HepG2和pcDNA3.1-IDO HepG2細(xì)胞分別為185. 60±15.58、198.80±12.60、269.00±21.60,前二者與后者比較差異具有統(tǒng)計(jì)學(xué)意義( P 0. 01)。 結(jié)論:IDO改變了HepG2細(xì)胞的生物學(xué)特點(diǎn),明顯促進(jìn)HepG2細(xì)胞增殖能力、黏附能力和體外侵襲能力。
[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 鹵0.18 and 1.07 鹵0.14, respectively, which was significantly lower than that of pcDNA3.1-IDO HepG (1.18 鹵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 鹵15.58198.80 鹵12.60269.00 鹵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.
【學(xué)位授予單位】：山西醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2011
【分類號(hào)】：R392

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