摘要
目的 探讨干扰素-α诱导肾癌细胞胸苷磷酸化酶（thymidine phosphorylase，TP）表达对5-氟尿嘧啶（5-FU）化疗敏感性的影响。方法采用不同浓度的干扰素-α2b处理肾癌细胞株786—0，应用RT—PCR和免疫印迹方法分别检测TP mRNA和TP蛋白水平变化。MTT法检测不同处理组786—0细胞中5-FU的半数抑制浓度（IC50）。建立肾透明细胞癌移植瘤动物模型，观察干扰素-α2b联合5-FU化疗对移植瘤生长的影响，免疫组化检测移植瘤中TP的表达，TUNEL检测肿瘤细胞的凋亡。结果 干扰素-α2b3000和6000IU／ml处理组，TP mRNA表达量（灰度峰值）分别为0．5733±0．0231和0．8233±0．0404，TP蛋白表达量分别为0．6347±0．0719和0．8735±0．0640。干扰素-α对TP的表达有剂量依赖性增强作用（P〈0．01）。在干扰素-α2b作用下，5-FU对786—0半数抑制浓度由（13．9467±3．7140）μmol／L下降至（5．3200±0．1039）μmol／L，化疗敏感性增加（P〈0．01）。联合用药组移植瘤体积为（0．0940±0．0492）cm^3，小于单纯5-FU组的（0．5424±0．1591）cm^3（P〈0．05）。单纯5-FU组和联合用药组移植瘤中肿瘤细胞凋亡指数差异无统计学意义（P〉0．05）。结论 干扰素-α2b诱导的TP增强表达参与了干扰素-α联合5-FU化疗增效作用，TP是干扰素-α2b的靶基因之。
Objective To explore the effect of thymidine phosphorylase expression induced by interferon-α on chemosensitivity of 5-Fluorouracil（5-FU） in renal carcinoma cell line. Methods Renal clear cell carcinoma cell line 786-0 was treated by various concentration of interferon（IFN）-α2b in vitro, and semi-quantitative RT-PCR was used to determine TP mRNA expression. Western blot was used to determine TP protein expression. 50% inhibitory concentration of growth（IC50） of 5-FU in different groups of treated 786-0 cell line was evaluated by MTT assay. The athymic mouse xenograft model of renal clear cell carcinoma were established, effect of chemotherapy of IFN-α2b combined with 5-FU was observed on the growth of xenograft tumor, and apoptosis and TP expression was observed by TUNEL and immunohistochemistry in xenograft tumor among different groups. Results In 3000 and 6000IU/ml IFN-α2b treated group, expression of TP mRNA were （0. 5733 ± 0. 0231） and （0. 8233±0. 0404）, and expression of TP protein were （0. 6347±0. 0719） and （0. 8735±0. 0640）. IFN-α2b has increased TP expression in a dose dependent manner（P〈0. 01 ）. IC50 of 5-FU to 786-0 cells was obviously decreased from （13. 9467±3. 7140）μmol/L to（5. 3200±0. 1039）μmol/L under the function of IFN-α2b,chemosensitivity of 5-FU was increased （P〈0.01）. Effect of chemotherapy of 5-FU was obviously enhanced in vivo after IFN-α2b combined with 5-FU and volume of xenograft tumor decreased from （0. 5424±0. 1591）cm^3 in 5-FU group to （0. 094±0. 0492）cm^3 in combination group （P〉0.05）. The apoptosis index were no different in xenograft of 5-FU and combination group（P〉 0.05）. Conclusions Up-modulating expression of TP induced by IFN-α has involved in enhanced cy totoxicity by combination use of 5-FU and IFN-α, TP is one of target genes of IFN-α.
出处
《中华泌尿外科杂志》
CAS
CSCD
北大核心
2007年第4期228-231,共4页
Chinese Journal of Urology
基金
河南省医学创新人才基金资助项目（200410733）
关键词
肾肿瘤
癌
胸苷磷酸化酶
干扰素
5-氟尿嘧啶
Kidney neoplasms
Carcinoma
Thymidine phosphorylase
Interferon
5-fluorouracil
