Xia Qiang, Wang Qi-wen, Zhang Ji-min
Department of Gastrointestinal Surgery, Guangzhou Medical University, Guangzhou, China.
Zhonghua Wei Chang Wai Ke Za Zhi. 2012 Jul;15(7):719-22.
To detect the effect of interferon-α2a(IFN-α2a) on thymidine phosphorylase(TP) mRNA expression levels and the anticancer activity of 5-fluorouracil(5-FU) and 5'-deoxy-fluorouridine(5'-DFUR) in human colon carcinoma cell lines LOVO and SW480.
Two human colon cancer cell lines LOVO and SW480 were cultured and treated with IFN-α2a at a series of dosage, and fluorescence quantitative PCR was carried out to detect the TP mRNA expression levels in these 2 cell lines. Then MTT assay and software Templet were used to determine the change of 50% inhibition concentration of 5-FU or 5'-DFUR combined with IFN-α2a on the two cell lines.
The TP mRNA expressions were up-regulated significantly by IFN-α2a at the doses of 500 U/ml and 5000 U/ml in LOVO(P<0.01). Compared with untreated cells(IFN-α2a 0 U/ml), no significance was found for TP mRNA expression levels in LOVO and SW480 treated by IFN-α2a at the dose of 50 U/ml (P>0.05). There was no significant difference for TP mRNA expression in SW480 between the dose of 0 U/ml and 500 U/ml of IFN-α2a(P>0.05), while a significant increace was detected at the dose of 5000 U/ml (P<0.01). No significant difference was found for the IC50 values after treatment of 5-FU combined with IFN-α2a (20 U/ml) on LOVO and SW480 compared with 5-FU alone, while the IC50 values after treatment of 5'-DFUR combined with IFN-α2a decreased significantly compared with 5'-DFUR alone(P<0.05).
There is no direct inhibition effect of IFN-α2a on LOVO and SW480 in vitro, while it can up-regulate TP mRNA expression levels both in LOVO and SW480, and enhance the anticancer effect of 5'-DFUR on these 2 cell lines.
检测α-2a干扰素(IFN-α2a)对人结肠癌LOVO和SW480细胞系中胸苷磷酸化酶(TP)mRNA表达水平及5-氟尿嘧啶(5-FU)和5'-脱氧氟尿苷(5'-DFUR)抗癌活性的影响。
培养两个人结肠癌细胞系LOVO和SW480,并用一系列剂量的IFN-α2a进行处理,然后进行荧光定量PCR以检测这两种细胞系中TP mRNA的表达水平。接着使用MTT法和软件Templet来确定5-FU或5'-DFUR与IFN-α2a联合作用于这两种细胞系时50%抑制浓度的变化。
在LOVO细胞中,500 U/ml和5000 U/ml剂量的IFN-α2a可显著上调TP mRNA表达(P<0.01)。与未处理细胞(IFN-α2a 0 U/ml)相比,50 U/ml剂量的IFN-α2a处理的LOVO和SW480细胞中TP mRNA表达水平无显著差异(P>0.05)。IFN-α2a 0 U/ml和500 U/ml剂量处理的SW480细胞中TP mRNA表达无显著差异(P>0.05),而5000 U/ml剂量时检测到显著增加(P<0.01)。与单独使用5-FU相比,5-FU与IFN-α2a(20 U/ml)联合处理LOVO和SW480细胞后的IC50值无显著差异,而5'-DFUR与IFN-α2a联合处理后的IC50值与单独使用5'-DFUR相比显著降低(P<0.05)。
IFN-α2a在体外对LOVO和SW480无直接抑制作用,但可上调LOVO和SW480细胞中TP mRNA表达水平,并增强5'-DFUR对这两种细胞系的抗癌作用。