目的研究外源性IL-18基因联合顺铂对胶质瘤C6细胞的凋亡诱导作用,为胶质瘤治疗提供新的途径。方法将IL-18基因及pLXSN病毒载体导入胶质瘤C6细胞,建立C6/IL-18及C6/pLXSN细胞;C6/IL-18、C6/pLX-SN及胶质瘤C6细胞均以RPMI1640培养基培养,经0.3、3、30、60、120μg/ml顺铂作用48h后,用酶标仪在490nm波长处测吸光度值,计算细胞抑制率;以顺铂的10倍血浆峰浓度作用(3μg/ml)细胞24h后,采用吖啶橙/溴乙啶(A//EB)双荧光染色法检测细胞凋亡率。结果 C6/IL-18细胞、C6/pLXSN细胞及亲代C6细胞经120、60、30、3、0.3μg/ml顺铂作用后的抑制率依次降低,C6/IL-18细胞抑制率明显高于C6/pLXSN和C6细胞(P均〈0.05)。以30μg/ml顺铂作用后C6/IL-18细胞的凋亡率明显高于C6/pLXSN和C6细胞(P均〈0.05)。结论外源性IL-18基因联合顺铂可明显增加对胶质瘤C6细胞的抑制作用,其相关机制有待进一步探讨。
Objective To explore the effect of exogenous IL-18 gene combined with cis-platinum on the induction of apoptosis in glioma C6 cell,and to develop new way for the therapy of glioma. Methods The glioma C6 cells were transducted with pLXSN retroviral vector containing( or not) IL-18 gene ,and C6 /IL-18 cells and C6 /pLXSN cells were prepared ; C6 /IL-18 cells,C6 /pLXSN cells and C6 cells were all cultured in RPMI1640 medium. After the cells were treated with cis-platinum at the concentration of 0. 3,3,30,60,120 μg/ml for 48 hs,the extinction values were measured using Microplate Reader at 490 nm wave length,and the inhibitory rates were calculated; the AO/EB double fluorescent staining was performed to detect the apoptotic rate after the cells exposed to the ten times peak serum concentration of cis-platinum for 24 hs. Results The inhibitory rate of C6 /IL-18 cells ,C6 /pLXSN cells and C6 cells after treated with 120,60,30,3 μg, 0. 3 μg/ml of cis-platinum decreased gradually,and which of the C6 /IL-18 cells were significantly higher than that of the C6 /pLXSN cells and C6 cells( P 0. 05) . The apoptotic rate of C6 /IL-18 cells after treated with 30 μg/ml of cis-platinum were significantly higher than that of the C6 /pLXSN cells and C6 cells( P 0. 05) . Conclusions Exogenous IL-18 gene combining with cis-platinum can enhance the inhibition of glioma C6 cell significantly,the mechanism of which maybe studied further.