塞来昔布联合携带白细胞介素24基因的溶瘤腺病毒对人类膀胱尿路上皮癌T24细胞的作用

Effects of celecoxib combined with oncolytic adenovirus armed with IL-24 gene on the human urothelial carcinoma T24 cells

目的评估塞来昔布联合携带白细胞介素24（IL-24）基因的溶瘤腺病毒（ZD55-IL-24）对人类膀胱尿路上皮癌T24细胞的选择性杀伤效能。方法荧光显微镜观察ZD55．增强型绿色荧光蛋白（EGFP）对细胞的转染效率。以人类脐静脉内皮细胞（HUVEC）作为对照。RT-PCR法测IL-24 mRNA在细胞中的表达。MTT法测塞来昔布联合ZD55-IL-24对肿瘤细胞选择性抑制作用。流式细胞术检测细胞凋亡情况。结果ZD55-EGFP感染细胞后，在相同时间下T24细胞中的荧光强度明显高于HUVEC细胞。ZD55-IL-24转染后在相同时间内T24细胞组IL-24 mRNA表达量均高于HUVEC组（t=-12．54，P〈0．01）。ZD55-IL-24与塞来昔布联合对T24细胞的抑制率最高（P〈0．01）。T24细胞各组的细胞增殖抑制率也明显高于HUVEC组（F=251．35，P〈0．01）。联合用药组的T24细胞凋亡率也明显高于单独用药组；T24细胞组凋亡率明显高于HUVEC组（P〈0．001）。结论塞来昔布与ZD55-IL-24联合用药能最大程度地抑制T24细胞的增殖，而对正常细胞影响轻微。这种抑制作用可能与细胞凋亡有关。

Objective To evaluate the ability to kill human urothelial carcinoma T24 cells selectively in vitro by celecoxib combined with ZD55-IL-24 and explore the effectiveness for this combination use. Methods The EGFP expression of cells was observed by fluorescence microscope. The human umbilical vein endothelial cells （HUVEC） were used as crotrols. The expression of IL-24 mRNA was detected by RT-PCR when the cells were transfected by ZD55-EGFP or ZD55-IL-24. After transfected by ZD55-IL-24 and treated by celecoxib, the inhibition effect on cells was measured by MTT assay, and the apoptosis rate was examined by flow cytometry. Results The fluorescence in T24 and HUVEC can be observed 24h after ZD55-EGFP transfection and the fluorescence intensity was increased corresponding with the times. Fluorescence intensity in T24 ceils showed higher than that in HUVEC group at the same times. The result of RT-PCR showed that the T24 cells expressed higher level of IL-24 mRNA than HUVEC group at the same time when the cells were transfect by ZD55-IL-24 （P 〈 0.01）. The inhibition rate of ZD55-IL-24 combined with celecoxib group was significantly higher than other groups （P 〈 0.001）. The inhibition rate of T24 cells in each group was significantly higher than HUVEC group （P 〈 0.01）. The flow cytometry results indicated that celecoxib combined with ZD55-IL-24 had the highest apoptosis rate on T24 cells than other single use group. Apoptosis rate of T24 ceils showed a higher than HUVEC cells （P 〈 0.01）. Conclusion Celecoxib combined with ZD55-IL-24 can inhibit T24 cells proliferation at a greatest degree and this effect may be contributed to apoptosis.