Cellular response to 5-fluorouracil (5-FU) in 5-FU-resistant colon cancer cell lines during treatment and recovery.
Cellular response to 5-fluorouracil (5-FU) in 5-FU-resistant colon cancer cell lines during treatment and recovery.
复制标题
在治疗和恢复过程中，对5-氟抗抗菌结肠癌细胞系中对5-氟尿嘧啶（5-FU）的细胞反应。
DOI：
10.1186/1476-4598-5-20
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发表时间：
2006-05-18
期刊：
影响因子：
37.3
通讯作者：
Stokke, Trond
中科院分区：
文献类型：
--
作者：
De Angelis, Paula M;Svendsrud, Debbie H;Kravik, Katherine L;Stokke, Trond
用抗癌药物5-氟尿嘧啶（5-FU）治疗细胞会导致DNA损伤，这反过来又会影响细胞增殖和存活。从HCT 116结肠癌细胞系产生的两种稳定的野生型TP 53 5-FU耐药细胞系，ContinB和ContinD，分别表现出对5-FU的中度和强烈耐药性，5-FU诱导的细胞凋亡水平显著降低，以及作为耐药性发展的结果的许多关键细胞周期和细胞凋亡调节基因的表达水平的改变。本研究的目的是确定这些耐药细胞系对8小时和24小时5-FU处理的潜在差异反应。我们评估了5-FU摄取到DNA中的水平，每个细胞系在整个治疗和恢复期的细胞周期效应和凋亡诱导，以及DNA损伤反应，细胞周期和凋亡调节基因表达水平的变化，以响应短期药物暴露。5-FU处理24小时导致S期阻滞，p53积累，p53靶基因在DNA损伤反应（ATF 3，GADD 34，GADD 45 A，PCNA），细胞周期调节（CDKN 1A）和凋亡调节途径（FAS）上的上调，以及亲本和耐药细胞系中的凋亡诱导。细胞系的5-FU掺入DNA的水平相似。恢复期细胞周期进程的模式一致地表明，5-FU耐药细胞系具有最小的S期分数和最大的G2（/M）分数。与其他两种细胞系相比，强5-FU耐药ContinD细胞系具有最小的S期停滞、最低的CDKN 1A水平和整个治疗和恢复期的最低水平的5-FU诱导的细胞凋亡，以及最快的指数生长恢复（10天）。中度5-FU耐药的ContinB细胞系在治疗和恢复期间的凋亡水平相对低于亲本细胞，恢复时间为22天。有丝分裂活动停止响应药物治疗的所有细胞系，有丝分裂调节基因的下调一致。证实了参与调节核苷酸结合/代谢（ATAD 2、GNL 2、GNL 3、MATR 3）、氨基酸代谢（AHCY、GSS、IVD、OAT）、细胞骨架组织（KRT 7、KRT 8、KRT 19、MAST 1）、转运（MTCH 1、NCBP 1、SNAPAP、VPS 52）和氧代谢（COX 5A、COX 7 C）的基因对5-FU处理的差异表达。我们的基因表达数据表明，核苷酸代谢，氨基酸代谢，细胞骨架组织，运输和氧代谢的调节改变可能是在这些细胞系中观察到的对5-FU的差异耐药的基础。在未来的研究中，将评估这些途径上一些受影响基因对5-FU耐药性的贡献作用。
Treatment of cells with the anti-cancer drug 5-fluorouracil (5-FU) causes DNA damage, which in turn affects cell proliferation and survival. Two stable wild-type TP53 5-FU-resistant cell lines, ContinB and ContinD, generated from the HCT116 colon cancer cell line, demonstrate moderate and strong resistance to 5-FU, respectively, markedly-reduced levels of 5-FU-induced apoptosis, and alterations in expression levels of a number of key cell cycle- and apoptosis-regulatory genes as a result of resistance development. The aim of the present study was to determine potential differential responses to 8 and 24-hour 5-FU treatment in these resistant cell lines. We assessed levels of 5-FU uptake into DNA, cell cycle effects and apoptosis induction throughout treatment and recovery periods for each cell line, and alterations in expression levels of DNA damage response-, cell cycle- and apoptosis-regulatory genes in response to short-term drug exposure. 5-FU treatment for 24 hours resulted in S phase arrests, p53 accumulation, up-regulation of p53-target genes on DNA damage response (ATF3, GADD34, GADD45A, PCNA), cell cycle-regulatory (CDKN1A), and apoptosis-regulatory pathways (FAS), and apoptosis induction in the parental and resistant cell lines. Levels of 5-FU incorporation into DNA were similar for the cell lines. The pattern of cell cycle progression during recovery demonstrated consistently that the 5-FU-resistant cell lines had the smallest S phase fractions and the largest G2(/M) fractions. The strongly 5-FU-resistant ContinD cell line had the smallest S phase arrests, the lowest CDKN1A levels, and the lowest levels of 5-FU-induced apoptosis throughout the treatment and recovery periods, and the fastest recovery of exponential growth (10 days) compared to the other two cell lines. The moderately 5-FU-resistant ContinB cell line had comparatively lower apoptotic levels than the parental cells during treatment and recovery periods and a recovery time of 22 days. Mitotic activity ceased in response to drug treatment for all cell lines, consistent with down-regulation of mitosis-regulatory genes. Differential expression in response to 5-FU treatment was demonstrated for genes involved in regulation of nucleotide binding/metabolism (ATAD2, GNL2, GNL3, MATR3), amino acid metabolism (AHCY, GSS, IVD, OAT), cytoskeleton organization (KRT7, KRT8, KRT19, MAST1), transport (MTCH1, NCBP1, SNAPAP, VPS52), and oxygen metabolism (COX5A, COX7C). Our gene expression data suggest that altered regulation of nucleotide metabolism, amino acid metabolism, cytoskeleton organization, transport, and oxygen metabolism may underlie the differential resistance to 5-FU seen in these cell lines. The contributory roles to 5-FU resistance of some of the affected genes on these pathways will be assessed in future studies.