CAJ | 학술논문

目的探讨卵巢特异启动子OSP-1调控下的白细胞介素(IL)-24基因对卵巢癌细胞系SKOV3的体外杀伤作用.方法应用基因重组技术构建OSP-1调控下的IL-24基因真核表达载体pcDNA3.0-OSP-1-IL-24,应用脂质体将该基因载体转染入人卵巢癌细胞SKOV3、人肝癌细胞HepG2及人成纤维细胞BJ.应用RT-PCR及ELISA方法检测IL-24基因在各组细胞内和培养上清中的表达情况,应用流式细胞术及M TT法检测IL-24基因对各组细胞杀伤作用.结果 pcDNA3.0-OSP-1-IL-24转染的3组细胞中,只有SKOV3细胞可检测到IL-24基因的表达,且只对SKOV3细胞有明显杀伤作用,而对其他两组细胞无杀伤作用.但OSP-1启动子系统杀伤SKOV3细胞的效能明显低于CMV启动子.MTT检测结果分析显示,转染pcDNA3.0-OSP-1-IL-24的SKOV3细胞培养上清对SKOV3细胞的生长有明显抑制作用.结论卵巢特异性启动子调控下的IL-24基因能够特异杀伤卵巢癌细胞,是一种新的卵巢癌靶向治疗方法.
목적 탐토란소특이계동자OSP-1조공하적백세포개소(IL)-24기인대란소암세포계SKOV3적체외살상작용.방법 응용기인중조기술구건OSP-1조공하적IL-24기인진핵표체재체pcDNA3.0-OSP-1-IL-24,응용지질체장해기인재체전염입인란소암세포SKOV3、인간암세포HepG2급인성섬유세포BJ.응용RT-PCR급ELISA방법검측IL-24기인재각조세포내화배양상청중적표체정황,응용류식세포술급M TT법검측IL-24기인대각조세포살상작용.결과 pcDNA3.0-OSP-1-IL-24전염적3조세포중,지유SKOV3세포가검측도IL-24기인적표체,차지대SKOV3세포유명현살상작용,이대기타량조세포무살상작용.단OSP-1계동자계통살상SKOV3세포적효능명현저우CMV계동자.MTT검측결과분석현시,전염pcDNA3.0-OSP-1-IL-24적SKOV3세포배양상청대SKOV3세포적생장유명현억제작용.결론 란소특이성계동자조공하적IL-24기인능구특이살상란소암세포,시일충신적란소암파향치료방법.
Objective To explore the in vitro antitumor effect of interleukin-24 (IL-24) gene regulated by ovarian-specific promoter-1 (OSP-1) on human ovarian cancer cell line SKOV3.Methods An expression vector (pcDNA3.0-OSP-1-IL-24) containing IL-24 gene under ovarian-specific promoter-1 was constructed by molecular biological methods and then transfected into human ovarian cancer cell SKOV3,human hepatoma cell HepG2 and human fibroblast BJ by cationic liposome.Reverse transcription-polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA) were used to detect the IL-24 gene in cells and supernatants.The antitumor effects of IL-24 gene were investigated by direct cell count,flow cytometry and methyl thiazolyl tetrazolium (MTT) assay.Results After transfections with pcDNA3.0-OSP-1-IL-24,the expression of IL-24 gene was detected only in SKOV3 cells.And only the growth of SKOV3 cells was inhibited significantly while those of HepG2 and BJ cells were not affected.The culture supernatant of SKOV3 cells transfected with pcDNA3.0-OSP-1-IL-24 could significantly inhibit the growth of SKOV3 cells.But the antitumor efficiency in SKOV3 cells of OSP-1 system was significantly lower than that of CMV promoter system.Conclusion With a high specificity,IL-24 gene therapy under the control of ovarian-specific promoter-1 is a novel target for the treatment of ovarian cancer.