摘要
目的探讨化疗药物5-氟尿嘧啶（5-Fu）在结肠癌细胞SW480和肝癌细胞HepG2对免疫共抑制分子B7-Ⅲ表达的诱导及机制。方法5-Fu处理细胞后24h用免疫荧光和流式细胞仪检测5-Fu对B7-H1蛋白表达的影响，在3～24h用荧光定量聚合酶链反应（RT-qPCR）检测其mRNA水平。结果5-Fu（10μg/L）处理这两种细胞24h后可探测出增强的B7-H1免疫荧光信号，流式细胞仪检测发现5-Fu处理SW480细胞后B7-H1蛋白含量为（14．88±1．43）％，处理HepG2细胞后B7-H1蛋白含量为（18．29±1．11）％，与各自对照组比较差异均有统计学意义（P〈0．05）。然而RT．qPCR检测表明在5．Fu处理后3～24h，SW480和HepG细胞内B7-H1mRNA的相对含量相对恒定。与各自对照组比较差异均无统计学意义（P〉0．05）。结论5-Fu在治疗肿瘤的同时会引起B7-Hl蛋白的表达，但这种增加与mRNA分子的转录无关。
Objective To study the mechanism of 5-fluorouracil （5-Fu） inducing B7-H1 in SW480 and HepG2 cells. Methods The B7-H1 protein expression was detected by using immunofluores- cence and flow cytometry in SW480 and HepG2 ceils at 24 h after 5-Fu treatment. The expression of B7-H1 mRNA was detected by using real-time quantitative polymerase chain reaction （RT-qPCR） 3-24 h after 5- Fu treatment. Results Immunofluorescence revealed a stronger fluorescence signal of B7-H1 in SW480 and HepG2 cells treated with 5-Fu （ 10 ixg/L） for 24 h. Flow cytometry demonstrated B7-H1 protein in SW480 ceils and HepG2 cells treated with 5-Fu treatment was （ 14. 88 -＋ 1.43）% and （ 18. 29 ＋ 1.11 ）% respectively, which was significantly different from that in control group （P 〈 0. 05 ）. Quantitative real-time PCR showed that the mRNA expression of B7-H1 was stable in SW480 and HepG2 cells 3-24 h after 5-Fu treatment. Conclusion 5-Fu could cause increased protein expression of B7-H1, which was not related with the increased mRNA expression.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2012年第12期2498-2499,共2页
Chinese Journal of Experimental Surgery
基金
湖北省自然科学基金重点资助项目（2011CDA065）
