摘要
目的将IL-18基因转染膀胱癌细胞(T24),检测该细胞体内外的凋亡变化,探讨IL-18的抗肿瘤作用机制。方法用含IL-18的基因IL-18/PA317转染T24细胞,经G418筛选后获得IL-18/T24阳性克隆。采用RT-PCR鉴定转染细胞mRNA的表达;用ELISA法检测IL-23/MA-891细胞培养上清中IL-18的分泌;用MTT比色法检测细胞体外增殖能力;观察IL-18/T24细胞生长情况、移植瘤的生长情况;用流式细胞技术检测体外和体内的细胞凋亡率。结果成功将IL-18基因转染T24细胞获得高表达IL-18细胞IL-18/T24。转染IL-18基因不影响T24细胞的体外生长和细胞凋亡,但体内IL-18基因转染组肿瘤生长明显受到抑制,肿瘤组织细胞凋亡率增高(P<0.01)。结论转染IL-18基因后对膀胱癌细胞(IL-18/T24)的体外凋亡无影响,但在体内IL-18能够诱导细胞凋亡,产生抗明显肿瘤作用。
Objective To set up bladder carcinoma cells (T24) transducted by IL-18 , and study its antitumor effect and related mechanism. Methods Transducted 124 cell with the culture supematant of IL-18/PA317 cells and gained IL-18/T24 cells selected with G418. IL-18 mRNA expression in IL-18/T24 cells was detected with RT-PCR. The expression of IL-18 protein was detected by ELISA. The proliferation of T24, LXSN/T24 and IL-18/T24 cells were detected by MTT colorimetry in vitro. Observe the growth of transplanted tumor in nude mouse. Flow cytometry was used to analyze the cell apoptosis.Results IL-18/T24 cells expressing IL-18 stably was successfully set up. The proliferation rate of IL-18/T24 cells showed no difference with parental cells, and the cell apoptosis of IL-18/T24 cells was similar in vitro( P 〉 0.05). Compared with the control group, growth rate of tumor in mice model was inhibited and cell apoptosis rate was increased significantly ( P 〈 0.01) in IL-18 gene therapy group. Conclusion The cellular clone IL-18/T24 with high level expression of IL-18 was produced and IL-18 has no effect on cell apoptosis in vitro, while it played a role in antitumor immunity by inducing cell apoptosis in vivo.
出处
《河北医药》
CAS
2008年第7期929-931,共3页
Hebei Medical Journal
基金
河北省科技厅科技攻关课题(编号:062761326)
关键词
IL-18
基因转染
膀胱癌细胞
抗肿瘤
凋亡
IL-18
gene transfection
bladder carcinoma cells
antitumor
apoptosis