Cellular response to 5-fluorouracil (5-FU) in 5-FU-resistant colon cancer cell lines during treatment and recovery
<p>Abstract</p> <p>Background</p> <p>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 <it>TP53 </it>5-FU-resistant cell lines, ContinB and Contin...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
| Published: |
BMC
2006-05-01
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| Series: | Molecular Cancer |
| Online Access: | http://www.molecular-cancer.com/content/5/1/20 |
| _version_ | 1830214440586313728 |
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| author | Kravik Katherine L Svendsrud Debbie H De Angelis Paula M Stokke Trond |
| author_facet | Kravik Katherine L Svendsrud Debbie H De Angelis Paula M Stokke Trond |
| author_sort | Kravik Katherine L |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>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 <it>TP53 </it>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.</p> <p>Results</p> <p>5-FU treatment for 24 hours resulted in S phase arrests, p53 accumulation, up-regulation of p53-target genes on DNA damage response (<it>ATF3</it>, <it>GADD34</it>, <it>GADD45A</it>, <it>PCNA</it>), cell cycle-regulatory (<it>CDKN1A</it>), and apoptosis-regulatory pathways (<it>FAS</it>), 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 G<sub>2</sub>(/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 (<it>ATAD2</it>, <it>GNL2</it>, <it>GNL3</it>, <it>MATR3</it>), amino acid metabolism (<it>AHCY</it>, <it>GSS</it>, <it>IVD</it>, <it>OAT</it>), cytoskeleton organization (<it>KRT7</it>, <it>KRT8</it>, <it>KRT19</it>, <it>MAST1</it>), transport (<it>MTCH1</it>, <it>NCBP1</it>, <it>SNAPAP</it>, <it>VPS52</it>), and oxygen metabolism (<it>COX5A</it>, <it>COX7C</it>).</p> <p>Conclusion</p> <p>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.</p> |
| first_indexed | 2024-12-18T06:42:48Z |
| format | Article |
| id | doaj.art-80898edf30084169aabb7ff2d63b3704 |
| institution | Directory Open Access Journal |
| issn | 1476-4598 |
| language | English |
| last_indexed | 2024-12-18T06:42:48Z |
| publishDate | 2006-05-01 |
| publisher | BMC |
| record_format | Article |
| series | Molecular Cancer |
| spelling | doaj.art-80898edf30084169aabb7ff2d63b37042022-12-21T21:17:35ZengBMCMolecular Cancer1476-45982006-05-01512010.1186/1476-4598-5-20Cellular response to 5-fluorouracil (5-FU) in 5-FU-resistant colon cancer cell lines during treatment and recoveryKravik Katherine LSvendsrud Debbie HDe Angelis Paula MStokke Trond<p>Abstract</p> <p>Background</p> <p>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 <it>TP53 </it>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.</p> <p>Results</p> <p>5-FU treatment for 24 hours resulted in S phase arrests, p53 accumulation, up-regulation of p53-target genes on DNA damage response (<it>ATF3</it>, <it>GADD34</it>, <it>GADD45A</it>, <it>PCNA</it>), cell cycle-regulatory (<it>CDKN1A</it>), and apoptosis-regulatory pathways (<it>FAS</it>), 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 G<sub>2</sub>(/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 (<it>ATAD2</it>, <it>GNL2</it>, <it>GNL3</it>, <it>MATR3</it>), amino acid metabolism (<it>AHCY</it>, <it>GSS</it>, <it>IVD</it>, <it>OAT</it>), cytoskeleton organization (<it>KRT7</it>, <it>KRT8</it>, <it>KRT19</it>, <it>MAST1</it>), transport (<it>MTCH1</it>, <it>NCBP1</it>, <it>SNAPAP</it>, <it>VPS52</it>), and oxygen metabolism (<it>COX5A</it>, <it>COX7C</it>).</p> <p>Conclusion</p> <p>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.</p>http://www.molecular-cancer.com/content/5/1/20 |
| spellingShingle | Kravik Katherine L Svendsrud Debbie H De Angelis Paula M Stokke Trond Cellular response to 5-fluorouracil (5-FU) in 5-FU-resistant colon cancer cell lines during treatment and recovery Molecular Cancer |
| title | Cellular response to 5-fluorouracil (5-FU) in 5-FU-resistant colon cancer cell lines during treatment and recovery |
| title_full | Cellular response to 5-fluorouracil (5-FU) in 5-FU-resistant colon cancer cell lines during treatment and recovery |
| title_fullStr | Cellular response to 5-fluorouracil (5-FU) in 5-FU-resistant colon cancer cell lines during treatment and recovery |
| title_full_unstemmed | Cellular response to 5-fluorouracil (5-FU) in 5-FU-resistant colon cancer cell lines during treatment and recovery |
| title_short | Cellular response to 5-fluorouracil (5-FU) in 5-FU-resistant colon cancer cell lines during treatment and recovery |
| title_sort | cellular response to 5 fluorouracil 5 fu in 5 fu resistant colon cancer cell lines during treatment and recovery |
| url | http://www.molecular-cancer.com/content/5/1/20 |
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