Genetic inactivation of the Fanconi anemia gene <it>FANCC </it>identified in the hepatocellular carcinoma cell line HuH-7 confers sensitivity towards DNA-interstrand crosslinking agents
<p>Abstract</p> <p>Background</p> <p>Inactivation of the Fanconi anemia (FA) pathway through defects in one of 13 FA genes occurs at low frequency in various solid cancer entities among the general population. As FA pathway inactivation confers a distinct hypersensitivi...
| Main Authors: | , , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2010-05-01
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| Series: | Molecular Cancer |
| Online Access: | http://www.molecular-cancer.com/content/9/1/127 |
| _version_ | 1818444061385687040 |
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| author | Bassermann Florian De Toni Enrico N Hanenberg Helmut Thasler Wolfgang E Meier Daniel Rizzani Antonia Ochs Stephanie Denk Gerald U Targosz Bianca-Sabrina Ziesch Andreas Plinninger Ursula Neveling Kornelia Palagyi Andreas Schäfer Claus Göke Burkhard Schindler Detlev Gallmeier Eike |
| author_facet | Bassermann Florian De Toni Enrico N Hanenberg Helmut Thasler Wolfgang E Meier Daniel Rizzani Antonia Ochs Stephanie Denk Gerald U Targosz Bianca-Sabrina Ziesch Andreas Plinninger Ursula Neveling Kornelia Palagyi Andreas Schäfer Claus Göke Burkhard Schindler Detlev Gallmeier Eike |
| author_sort | Bassermann Florian |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>Inactivation of the Fanconi anemia (FA) pathway through defects in one of 13 FA genes occurs at low frequency in various solid cancer entities among the general population. As FA pathway inactivation confers a distinct hypersensitivity towards DNA interstrand-crosslinking (ICL)-agents, FA defects represent rational targets for individualized therapeutic strategies. Except for pancreatic cancer, however, the prevalence of FA defects in gastrointestinal (GI) tumors has not yet been systematically explored.</p> <p>Results</p> <p>A panel of GI cancer cell lines was screened for FA pathway inactivation applying FANCD2 monoubiquitination and FANCD2/RAD51 nuclear focus formation and a newly identified FA pathway-deficient cell line was functionally characterized. The hepatocellular carcinoma (HCC) line HuH-7 was defective in FANCD2 monoubiquitination and FANCD2 nuclear focus formation but proficient in RAD51 focus formation. Gene complementation studies revealed that this proximal FA pathway inactivation was attributable to defective FANCC function in HuH-7 cells. Accordingly, a homozygous inactivating <it>FANCC </it>nonsense mutation (c.553C > T, p.R185X) was identified in HuH-7, resulting in partial transcriptional skipping of exon 6 and leading to the classic cellular FA hypersensitivity phenotype; HuH-7 cells exhibited a strongly reduced proliferation rate and a pronounced G2 cell cycle arrest at distinctly lower concentrations of ICL-agents than a panel of non-isogenic, FA pathway-proficient HCC cell lines. Upon retroviral transduction of HuH-7 cells with <it>FANCC </it>cDNA, FA pathway functions were restored and ICL-hypersensitivity abrogated. Analyses of 18 surgical HCC specimens yielded no further examples for genetic or epigenetic inactivation of <it>FANCC</it>, <it>FANCF</it>, or <it>FANCG </it>in HCC, suggesting a low prevalence of proximal FA pathway inactivation in this tumor type.</p> <p>Conclusions</p> <p>As the majority of HCC are chemoresistant, assessment of FA pathway function in HCC could identify small subpopulations of patients expected to predictably benefit from individualized treatment protocols using ICL-agents.</p> |
| first_indexed | 2024-12-14T19:09:57Z |
| format | Article |
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| institution | Directory Open Access Journal |
| issn | 1476-4598 |
| language | English |
| last_indexed | 2024-12-14T19:09:57Z |
| publishDate | 2010-05-01 |
| publisher | BMC |
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| series | Molecular Cancer |
| spelling | doaj.art-4103240894fe4ec7af00b429a1fdf69a2022-12-21T22:50:44ZengBMCMolecular Cancer1476-45982010-05-019112710.1186/1476-4598-9-127Genetic inactivation of the Fanconi anemia gene <it>FANCC </it>identified in the hepatocellular carcinoma cell line HuH-7 confers sensitivity towards DNA-interstrand crosslinking agentsBassermann FlorianDe Toni Enrico NHanenberg HelmutThasler Wolfgang EMeier DanielRizzani AntoniaOchs StephanieDenk Gerald UTargosz Bianca-SabrinaZiesch AndreasPlinninger UrsulaNeveling KorneliaPalagyi AndreasSchäfer ClausGöke BurkhardSchindler DetlevGallmeier Eike<p>Abstract</p> <p>Background</p> <p>Inactivation of the Fanconi anemia (FA) pathway through defects in one of 13 FA genes occurs at low frequency in various solid cancer entities among the general population. As FA pathway inactivation confers a distinct hypersensitivity towards DNA interstrand-crosslinking (ICL)-agents, FA defects represent rational targets for individualized therapeutic strategies. Except for pancreatic cancer, however, the prevalence of FA defects in gastrointestinal (GI) tumors has not yet been systematically explored.</p> <p>Results</p> <p>A panel of GI cancer cell lines was screened for FA pathway inactivation applying FANCD2 monoubiquitination and FANCD2/RAD51 nuclear focus formation and a newly identified FA pathway-deficient cell line was functionally characterized. The hepatocellular carcinoma (HCC) line HuH-7 was defective in FANCD2 monoubiquitination and FANCD2 nuclear focus formation but proficient in RAD51 focus formation. Gene complementation studies revealed that this proximal FA pathway inactivation was attributable to defective FANCC function in HuH-7 cells. Accordingly, a homozygous inactivating <it>FANCC </it>nonsense mutation (c.553C > T, p.R185X) was identified in HuH-7, resulting in partial transcriptional skipping of exon 6 and leading to the classic cellular FA hypersensitivity phenotype; HuH-7 cells exhibited a strongly reduced proliferation rate and a pronounced G2 cell cycle arrest at distinctly lower concentrations of ICL-agents than a panel of non-isogenic, FA pathway-proficient HCC cell lines. Upon retroviral transduction of HuH-7 cells with <it>FANCC </it>cDNA, FA pathway functions were restored and ICL-hypersensitivity abrogated. Analyses of 18 surgical HCC specimens yielded no further examples for genetic or epigenetic inactivation of <it>FANCC</it>, <it>FANCF</it>, or <it>FANCG </it>in HCC, suggesting a low prevalence of proximal FA pathway inactivation in this tumor type.</p> <p>Conclusions</p> <p>As the majority of HCC are chemoresistant, assessment of FA pathway function in HCC could identify small subpopulations of patients expected to predictably benefit from individualized treatment protocols using ICL-agents.</p>http://www.molecular-cancer.com/content/9/1/127 |
| spellingShingle | Bassermann Florian De Toni Enrico N Hanenberg Helmut Thasler Wolfgang E Meier Daniel Rizzani Antonia Ochs Stephanie Denk Gerald U Targosz Bianca-Sabrina Ziesch Andreas Plinninger Ursula Neveling Kornelia Palagyi Andreas Schäfer Claus Göke Burkhard Schindler Detlev Gallmeier Eike Genetic inactivation of the Fanconi anemia gene <it>FANCC </it>identified in the hepatocellular carcinoma cell line HuH-7 confers sensitivity towards DNA-interstrand crosslinking agents Molecular Cancer |
| title | Genetic inactivation of the Fanconi anemia gene <it>FANCC </it>identified in the hepatocellular carcinoma cell line HuH-7 confers sensitivity towards DNA-interstrand crosslinking agents |
| title_full | Genetic inactivation of the Fanconi anemia gene <it>FANCC </it>identified in the hepatocellular carcinoma cell line HuH-7 confers sensitivity towards DNA-interstrand crosslinking agents |
| title_fullStr | Genetic inactivation of the Fanconi anemia gene <it>FANCC </it>identified in the hepatocellular carcinoma cell line HuH-7 confers sensitivity towards DNA-interstrand crosslinking agents |
| title_full_unstemmed | Genetic inactivation of the Fanconi anemia gene <it>FANCC </it>identified in the hepatocellular carcinoma cell line HuH-7 confers sensitivity towards DNA-interstrand crosslinking agents |
| title_short | Genetic inactivation of the Fanconi anemia gene <it>FANCC </it>identified in the hepatocellular carcinoma cell line HuH-7 confers sensitivity towards DNA-interstrand crosslinking agents |
| title_sort | genetic inactivation of the fanconi anemia gene it fancc it identified in the hepatocellular carcinoma cell line huh 7 confers sensitivity towards dna interstrand crosslinking agents |
| url | http://www.molecular-cancer.com/content/9/1/127 |
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