CAJ | 학술논문

目的：构建人谷胱甘肽过氧化物酶2（Glutathione peroxidase 2, GPX2）慢病毒干涉载体，获得稳定下调GPX2的高滴度慢病毒，检测敲低GPX2对细胞凋亡的影响。方法通过筛选具有显著干涉效果的siRNA序列，构建pSicoR-GPX2慢病毒干涉载体，包装病毒并感染人肝癌HepG2细胞，分别用Western-blot和RT-PCR方法检测GPX2表达情况，应用流式细胞术分析敲低GPX2对人肝癌细胞凋亡的影响。结果成功构建pSicoR-GPX2干涉慢病毒载体并获得高滴度慢病毒颗粒，GPX2蛋白表达水平和RNA表达水平均有显著下调，且人肝癌细胞感染GPX2干涉慢病毒后对凋亡更加敏感，其原因可能是促凋亡蛋白Bax的活化和抗凋亡蛋白Bcl-2活性的抑制。结论成功构建人GPX2基因干涉慢病毒，为肝癌靶标治疗提供新的线索，为后续GPX2功能研究奠定基础。
목적：구건인곡광감태과양화물매2（Glutathione peroxidase 2, GPX2）만병독간섭재체，획득은정하조GPX2적고적도만병독，검측고저GPX2대세포조망적영향。방법통과사선구유현저간섭효과적siRNA서렬，구건pSicoR-GPX2만병독간섭재체，포장병독병감염인간암HepG2세포，분별용Western-blot화RT-PCR방법검측GPX2표체정황，응용류식세포술분석고저GPX2대인간암세포조망적영향。결과성공구건pSicoR-GPX2간섭만병독재체병획득고적도만병독과립，GPX2단백표체수평화RNA표체수평균유현저하조，차인간암세포감염GPX2간섭만병독후대조망경가민감，기원인가능시촉조망단백Bax적활화화항조망단백Bcl-2활성적억제。결론성공구건인GPX2기인간섭만병독，위간암파표치료제공신적선색，위후속GPX2공능연구전정기출。
Objective To construct a RNA interference lentiviral vector for human glutathione peroxidase 2 (GPX2) gene and observe the effect of GPX2 knockdown on cell apoptosis. Methods The sequence of the small interfering RNA (siRNA) for GPX2 interference was inserted into the pSicoR vector. HepG2 cells were infected by the packaged si-GPX2 lentivirus and the expression of GPX2 in the infected cells was detected by both RT-PCR and Western blotting. Changes of cell apoptosis following the infection were analyzed by flow cytometry. Results The lentiviral particles pSicoR-GPX2 were successfully packaged. The expression of GPX2 in the infected cells was obviously down-regulated at both RNA and protein levels. GPX2 knockdown caused increased apoptosis rate, increased Bax expression and lowered Bcl-2 expression in HepG2 cells. Conclusion We have successfully constructed the lentiviral vector for RNA interference of human GPX2 gene.