{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T03:16:07Z","timestamp":1790738167830,"version":"4.1.0"},"reference-count":55,"publisher":"Oxford University Press (OUP)","issue":"6","license":[{"start":{"date-parts":[[2017,6,1]],"date-time":"2017-06-01T00:00:00Z","timestamp":1496275200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/pages\/standard-publication-reuse-rights"}],"content-domain":{"domain":["rep.bioscientifica.com"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[2017,6,1]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Anticancer drugs, such as alkylating agents, can affect male fertility by targeting the DNA of proliferative spermatogonial stem cells (SSC). Therefore, to reduce such side effects, other chemotherapeutics are used. However, less is known about their potential genotoxicity on SSC. Moreover, DNA repair mechanisms in SSC are poorly understood. To model treatments deprived of alkylating agents that are commonly used in cancer treatment, we tested the impact of exposure to doxorubicin and vincristine, alone or in combination (MIX), on a rat spermatogonial cell line with SSC characteristics (GC-6spg). Vincristine alone induced a cell cycle arrest and cell death without genotoxic impact. On the other hand, doxorubicin and the MIX induced a dose-dependent cell death. More importantly, doxorubicin and the MIX induced DNA breaks, measured by the COMET assay, at a non-cytotoxic dose. To elucidate which DNA repair pathway is activated in spermatogonia after exposure to doxorubicin, we screened the expression of 75 genes implicated in DNA repair. Interestingly, all were expressed constitutively in GC-6spg, suggesting great potential to respond to genotoxic stress. Doxorubicin treatments affected the expression of 16 genes (&amp;gt;1.5 fold change; P\u2009&amp;lt;\u20090.05) involved in cell cycle, base\/nucleotide excision repair, homologous recombination and non-homologous end joining (NHEJ). The significant increase in CDKN1A and XRCC1 suggest a cell cycle arrest and implies an alternative NHEJ pathway in response to doxorubicin-induced DNA breaks. Together, our results support the idea that undifferentiated spermatogonia have the ability to respond to DNA injury from chemotherapeutic compounds and escape DNA break accumulation.<\/jats:p>","DOI":"10.1530\/rep-17-0005","type":"journal-article","created":{"date-parts":[[2017,3,3]],"date-time":"2017-03-03T20:50:13Z","timestamp":1488574213000},"page":"725-735","update-policy":"https:\/\/doi.org\/10.1530\/crossmarkpolicy-1","source":"Crossref","is-referenced-by-count":31,"title":["Doxorubicin and vincristine affect undifferentiated rat spermatogonia"],"prefix":"10.1093","volume":"153","author":[{"given":"Hermance","family":"Beaud","sequence":"first","affiliation":[{"name":"INRS-Institut Armand Frappier \u00a0 Laval, Quebec, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ans","family":"van Pelt","sequence":"additional","affiliation":[{"name":"Center for Reproductive Medicine \u00a0 Academic Medical Center, University of Amsterdam, Amsterdam, the Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Geraldine","family":"Delbes","sequence":"additional","affiliation":[{"name":"INRS-Institut Armand Frappier \u00a0 Laval, Quebec, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2017,6,1]]},"reference":[{"key":"2026010712132492600_bib1","doi-asserted-by":"crossref","first-page":"1024","DOI":"10.1095\/biolreprod66.4.1024","article-title":"Chronic cyclophosphamide treatment alters the expression of stress response genes in rat male germ cells","volume":"66","author":"Aguilar-Mahecha","year":"2002","journal-title":"Biology of Reproduction"},{"key":"2026010712132492600_bib2","first-page":"572","article-title":"Distribution and metabolism of doxorubicin in rats undergoing testicular circulatory isolation","volume":"162","author":"Arakawa","year":"1991","journal-title":"American Journal of Surgical Pathology"},{"key":"2026010712132492600_bib3","doi-asserted-by":"crossref","first-page":"55117","DOI":"10.1074\/jbc.M404524200","article-title":"Involvement of poly(ADP-ribose) polymerase-1 and XRCC1\/DNA ligase III in an alternative route for DNA double-strand breaks rejoining","volume":"279","author":"Audebert","year":"2004","journal-title":"Journal of Biological Chemistry"},{"key":"2026010712132492600_bib4","doi-asserted-by":"crossref","first-page":"667.e663","DOI":"10.1016\/j.fertnstert.2014.06.008","article-title":"Impact of lymphoma treatments on spermatogenesis and sperm deoxyribonucleic acid: a multicenter prospective study from the CECOS network","volume":"102","author":"Bujan","year":"2014","journal-title":"Fertility and Sterility"},{"key":"2026010712132492600_bib5","doi-asserted-by":"crossref","first-page":"234","DOI":"10.1093\/toxsci\/kfm160","article-title":"Developmental changes in testicular sensitivity to estrogens throughout fetal and neonatal life","volume":"99","author":"Delbes","year":"2007","journal-title":"Toxicological Sciences"},{"key":"2026010712132492600_bib6","doi-asserted-by":"crossref","first-page":"241","DOI":"10.2164\/jandrol.106.001487","article-title":"Effects of the chemotherapy cocktail used to treat testicular cancer on sperm chromatin integrity.","volume":"28","author":"Delbes","year":"2007","journal-title":"Journal of Andrology"},{"key":"2026010712132492600_bib7","doi-asserted-by":"crossref","first-page":"14","DOI":"10.1093\/molehr\/gap087","article-title":"Toxicants and human sperm chromatin integrity","volume":"16","author":"Delbes","year":"2010","journal-title":"Molecular Human Reproduction"},{"key":"2026010712132492600_bib8","doi-asserted-by":"crossref","first-page":"332","DOI":"10.1016\/j.reprotox.2010.01.007","article-title":"Reversibility of the effects of the chemotherapeutic regimen for non-Hodgkin lymphoma, cyclophosphamide, doxorubicin, vincristine, and prednisone, on the male rat reproductive system and progeny outcome","volume":"29","author":"Delbes","year":"2010","journal-title":"Reproductive Toxicology"},{"key":"2026010712132492600_bib9","doi-asserted-by":"crossref","first-page":"24","DOI":"10.1016\/j.reprotox.2012.10.012","article-title":"Selective induction of glutathione S-transferases in round spermatids from the Brown-Norway rat by the chemotherapeutic regimen for testicular cancer","volume":"36","author":"Delbes","year":"2013","journal-title":"Reproductive Toxicology"},{"key":"2026010712132492600_bib10","doi-asserted-by":"crossref","first-page":"395","DOI":"10.1016\/j.reprotox.2011.09.010","article-title":"Effects of multiple doses of cyclophosphamide on mouse testes: accessing the germ cells lost, and the functional damage of stem cells","volume":"32","author":"Drumond","year":"2011","journal-title":"Reproductive Toxicology"},{"key":"2026010712132492600_bib11","doi-asserted-by":"crossref","first-page":"40","DOI":"10.1128\/MMBR.00045-13","article-title":"DNA repair pathways in trypanosomatids: from DNA repair to drug resistance","volume":"78","author":"Genois","year":"2014","journal-title":"Microbiology and Molecular Biology Reviews"},{"key":"2026010712132492600_bib12","doi-asserted-by":"crossref","first-page":"1215","DOI":"10.1016\/S1470-2045(14)70408-5","article-title":"Cumulative alkylating agent exposure and semen parameters in adult survivors of childhood cancer: a report from the St Jude Lifetime Cohort study","volume":"15","author":"Green","year":"2014","journal-title":"Lancet Oncology"},{"key":"2026010712132492600_bib13","doi-asserted-by":"crossref","first-page":"70","DOI":"10.1016\/j.toxlet.2016.11.016","article-title":"Germ cell responses to doxorubicin exposure in vitro","volume":"265","author":"Habas","year":"2016","journal-title":"Toxicology Letters"},{"key":"2026010712132492600_bib14","doi-asserted-by":"crossref","first-page":"717","DOI":"10.1095\/biolreprod.102.008920","article-title":"Function of DNA-protein kinase catalytic subunit during the early meiotic prophase without Ku70 and Ku86","volume":"68","author":"Hamer","year":"2003","journal-title":"Biology of Reproduction"},{"key":"2026010712132492600_bib15","doi-asserted-by":"crossref","first-page":"133","DOI":"10.1007\/s00280-001-0392-4","article-title":"Peak plasma concentrations of doxorubicin in children with acute lymphoblastic leukemia or non-Hodgkin lymphoma","volume":"49","author":"Hempel","year":"2002","journal-title":"Cancer Chemotherapy and Pharmacology"},{"key":"2026010712132492600_bib16","doi-asserted-by":"crossref","first-page":"1941","DOI":"10.1200\/JCO.2013.54.0914","article-title":"Prevention and management of chemotherapy-induced peripheral neuropathy in survivors of adult cancers: American Society of Clinical Oncology Clinical Practice Guideline","volume":"32","author":"Hershman","year":"2014","journal-title":"Journal of Clinical Oncology"},{"key":"2026010712132492600_bib17","doi-asserted-by":"crossref","first-page":"9999","DOI":"10.1158\/0008-5472.CAN-05-2004","article-title":"Doxorubicin induces apoptosis in germ line stem cells in the immature rat testis and amifostine cannot protect against this cytotoxicity","volume":"65","author":"Hou","year":"2005","journal-title":"Cancer Research"},{"key":"2026010712132492600_bib18","doi-asserted-by":"crossref","first-page":"12","DOI":"10.1093\/jncimonographs\/lgi003","article-title":"Spermatogenesis after cancer treatment: damage and recovery","volume":"34","author":"Howell","year":"2005","journal-title":"Journal of the National Cancer Institute Monographs"},{"key":"2026010712132492600_bib19","doi-asserted-by":"crossref","first-page":"147","DOI":"10.1016\/j.bcmd.2013.12.005","article-title":"DNA damage response in adult stem cells","volume":"52","author":"Insinga","year":"2014","journal-title":"Blood Cells, Molecules, and Diseases"},{"key":"2026010712132492600_bib20","doi-asserted-by":"crossref","first-page":"2410","DOI":"10.1128\/MCB.22.7.2410-2418.2002","article-title":"Base excision repair is limited by different proteins in male germ cell nuclear extracts prepared from young and old mice","volume":"22","author":"Intano","year":"2002","journal-title":"Molecular and Cellular Biology"},{"key":"2026010712132492600_bib21","doi-asserted-by":"crossref","first-page":"482","DOI":"10.1095\/biolreprod63.2.482","article-title":"Stage-specific inhibition of deoxyribonucleic acid synthesis and induction of apoptosis by antracyclines in cultured rat spermatogenic cells","volume":"63","author":"Jahnukainen","year":"2000","journal-title":"Biology of Reproduction"},{"key":"2026010712132492600_bib22","doi-asserted-by":"crossref","first-page":"253","DOI":"10.1038\/nrc1317","article-title":"Microtubules as a target for anticancer drugs","volume":"4","author":"Jordan","year":"2004","journal-title":"Nature Reviews Cancer"},{"key":"2026010712132492600_bib23","doi-asserted-by":"crossref","first-page":"7749","DOI":"10.1038\/sj.onc.1210880","article-title":"ATM and the Mre11 complex combine to recognize and signal DNA double-strand breaks","volume":"26","author":"Lavin","year":"2007","journal-title":"Oncogene"},{"key":"2026010712132492600_bib24","doi-asserted-by":"crossref","first-page":"72","DOI":"10.1095\/biolreprod.114.117754","article-title":"Effects of four chemotherapeutic agents, bleomycin, etoposide, cisplatin, and cyclophosphamide, on DNA damage and telomeres in a mouse spermatogonial cell line","volume":"90","author":"Liu","year":"2014","journal-title":"Biology of Reproduction"},{"key":"2026010712132492600_bib25","doi-asserted-by":"crossref","first-page":"228","DOI":"10.1095\/biolreprod.110.083568","article-title":"Development of a short-term fluorescence-based assay to assess the toxicity of anticancer drugs on rat stem\/progenitor spermatogonia in vitro","volume":"83","author":"Marcon","year":"2010","journal-title":"Biology of Reproduction"},{"key":"2026010712132492600_bib26","doi-asserted-by":"crossref","first-page":"432","DOI":"10.2164\/jandrol.110.011601","article-title":"Effects of chemotherapeutic agents for testicular cancer on rat spermatogonial stem\/progenitor cells","volume":"32","author":"Marcon","year":"2011","journal-title":"Journal of Andrology"},{"key":"2026010712132492600_bib27","first-page":"89","article-title":"Relationship between spermatogonial stem cell survival and testis function after cytotoxic therapy","volume":"7","author":"Meistrich","year":"1986","journal-title":"British Journal of Cancer"},{"key":"2026010712132492600_bib28","doi-asserted-by":"crossref","first-page":"261","DOI":"10.1002\/pbc.22004","article-title":"Male gonadal toxicity","volume":"53","author":"Meistrich","year":"2009","journal-title":"Pediatric Blood and Cancer"},{"key":"2026010712132492600_bib29","doi-asserted-by":"crossref","first-page":"1180","DOI":"10.1016\/j.fertnstert.2013.08.010","article-title":"Effects of chemotherapy and radiotherapy on spermatogenesis in humans","volume":"100","author":"Meistrich","year":"2013","journal-title":"Fertility and Sterility"},{"key":"2026010712132492600_bib30","doi-asserted-by":"crossref","first-page":"2703","DOI":"10.1002\/1097-0142(19921201)70:11<2703::AID-CNCR2820701123>3.0.CO;2-X","article-title":"Impact of cyclophosphamide on long-term reduction in sperm count in men treated with combination chemotherapy for ewing and soft-tissue sarcomas","volume":"70","author":"Meistrich","year":"1992","journal-title":"Cancer"},{"key":"2026010712132492600_bib31","first-page":"1","article-title":"Pharmacokinetics of adriamycin after intravenous administration in rat","volume":"6","author":"Nwankwoala","year":"2008","journal-title":"Internet Journal of Oncology"},{"key":"2026010712132492600_bib32","doi-asserted-by":"crossref","first-page":"1374","DOI":"10.1016\/j.fertnstert.2009.05.068","article-title":"Impact of chemotherapeutics and advanced testicular cancer or Hodgkin lymphoma on sperm deoxyribonucleic acid integrity","volume":"94","author":"O\u2019Flaherty","year":"2010","journal-title":"Fertility and Sterility"},{"key":"2026010712132492600_bib33","doi-asserted-by":"crossref","first-page":"629","DOI":"10.2164\/jandrol.111.015388","article-title":"Sperm chromatin structure components are differentially repaired in cancer survivors","volume":"33","author":"O\u2019Flaherty","year":"2012","journal-title":"Journal of Andrology"},{"key":"2026010712132492600_bib34","first-page":"505","article-title":"DNA repair mechanisms in mammalian germ cells","volume":"26","author":"Ozturk","year":"2011","journal-title":"Histology and Histopathology"},{"key":"2026010712132492600_bib35","doi-asserted-by":"crossref","first-page":"789","DOI":"10.1095\/biolreprod62.3.789","article-title":"Expression of deoxyribonucleic acid repair enzymes during spermatogenesis in mice","volume":"62","author":"Richardson","year":"2000","journal-title":"Biology of Reproduction"},{"key":"2026010712132492600_bib36","article-title":"Cell viability assays","volume-title":"Assay Guidance Manual","author":"Riss","year":"2004"},{"key":"2026010712132492600_bib37","doi-asserted-by":"crossref","first-page":"326","DOI":"10.1016\/j.tips.2015.03.005","article-title":"Anthracyclines\/trastuzumab: new aspects of cardiotoxicity and molecular mechanisms","volume":"36","author":"Rochette","year":"2015","journal-title":"Trends in Pharmacological Sciences"},{"key":"2026010712132492600_bib38","doi-asserted-by":"crossref","first-page":"5706","DOI":"10.1128\/MCB.23.16.5706-5715.2003","article-title":"Pathways of DNA double-strand break repair during the mammalian cell cycle","volume":"23","author":"Rothkamm","year":"2003","journal-title":"Molecular and Cellular Biology"},{"key":"2026010712132492600_bib39","doi-asserted-by":"crossref","first-page":"159","DOI":"10.1016\/j.dnarep.2010.10.007","article-title":"Protecting the heritable genome: DNA damage response mechanisms in spermatogonial stem cells","volume":"10","author":"Rube","year":"2011","journal-title":"DNA Repair"},{"key":"2026010712132492600_bib40","doi-asserted-by":"crossref","first-page":"111","DOI":"10.1007\/BF00262326","article-title":"Pharmacokinetics of vincristine sulfate in children","volume":"6","author":"Sethi","year":"1981","journal-title":"Cancer Chemotherapy and Pharmacology"},{"key":"2026010712132492600_bib41","doi-asserted-by":"crossref","first-page":"1276","DOI":"10.1021\/jm960835d","article-title":"Redox pathway leading to the alkylation of DNA by the anthracycline, antitumor drugs adriamycin and daunomycin","volume":"40","author":"Taatjes","year":"1997","journal-title":"Journal of Medicinal Chemistry"},{"key":"2026010712132492600_bib42","doi-asserted-by":"crossref","first-page":"170","DOI":"10.1016\/j.stem.2007.10.023","article-title":"Stem cell defects in ATM-deficient undifferentiated spermatogonia through DNA damage-induced cell-cycle arrest","volume":"2","author":"Takubo","year":"2008","journal-title":"Cell Stem Cell"},{"key":"2026010712132492600_bib43","doi-asserted-by":"crossref","first-page":"251","DOI":"10.1093\/humrep\/dem389","article-title":"Sperm aneuploidy frequencies analysed before and after chemotherapy in testicular cancer and Hodgkin\u2019s lymphoma patients","volume":"23","author":"Tempest","year":"2008","journal-title":"Human Reproduction"},{"key":"2026010712132492600_bib44","doi-asserted-by":"crossref","first-page":"611","DOI":"10.1093\/humrep\/deh127","article-title":"Late, but not early, paternal effect on human embryo development is related to sperm DNA fragmentation","volume":"19","author":"Tesarik","year":"2004","journal-title":"Human Reproduction"},{"key":"2026010712132492600_bib45","first-page":"339","article-title":"Role of O6-alkylguanine-DNA alkyltransferase in the resistance of mouse spermatogenic cells to O6-alkylating agents","volume":"119","author":"Thompson","year":"2000","journal-title":"Journal of Reproduction and Infertility"},{"key":"2026010712132492600_bib46","doi-asserted-by":"crossref","first-page":"1100","DOI":"10.1016\/j.dnarep.2007.03.011","article-title":"DNA binding, nucleotide flipping, and the helix-turn-helix motif in base repair by O6-alkylguanine-DNA alkyltransferase and its implications for cancer chemotherapy","volume":"6","author":"Tubbs","year":"2007","journal-title":"DNA Repair"},{"key":"2026010712132492600_bib47","doi-asserted-by":"crossref","first-page":"578","DOI":"10.2164\/jandrol.106.002428","article-title":"Effects of the chemotherapeutic agents for non-Hodgkin lymphoma, cyclophosphamide, doxorubicin, vincristine, and prednisone (CHOP), on the male rat reproductive system and progeny outcome","volume":"28","author":"Vaisheva","year":"2007","journal-title":"Journal of Andrology"},{"key":"2026010712132492600_bib48","doi-asserted-by":"crossref","first-page":"1845","DOI":"10.1210\/endo.143.5.8806","article-title":"Establishment of cell lines with rat spermatogonial stem cell characteristics","volume":"143","author":"Van Pelt","year":"2002","journal-title":"Endocrinology"},{"key":"2026010712132492600_bib49","doi-asserted-by":"crossref","first-page":"3204","DOI":"10.3390\/biom5043204","article-title":"DNA Damage signalling and repair inhibitors: the long-sought-after achilles\u2019 heel of cancer","volume":"5","author":"Velic","year":"2015","journal-title":"Biomolecules"},{"key":"2026010712132492600_bib50","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1186\/1477-7827-8-3","article-title":"Amifostine reduces the seminiferous epithelium damage in doxorubicin-treated prepubertal rats without improving the fertility status","volume":"8","author":"Vendramini","year":"2010","journal-title":"Reproductive Biology and Endocrinology"},{"key":"2026010712132492600_bib51","doi-asserted-by":"crossref","first-page":"2457","DOI":"10.1093\/humrep\/des159","article-title":"Amifostine-doxorubicin association causes long-term prepubertal spermatogonia DNA damage and early developmental arrest","volume":"27","author":"Vendramini","year":"2012","journal-title":"Human Reproduction"},{"key":"2026010712132492600_bib52","doi-asserted-by":"crossref","first-page":"209","DOI":"10.1016\/S1470-2045(05)70092-9","article-title":"Fertility preservation for young patients with cancer: who is at risk and what can be offered?","volume":"6","author":"Wallace","year":"2005","journal-title":"Lancet Oncology"},{"key":"2026010712132492600_bib53","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1095\/biolreprod.104.039123","article-title":"Nucleotide excision repair activity varies among murine spermatogenic cell types","volume":"73","author":"Xu","year":"2005","journal-title":"Biology of Reproduction"},{"key":"2026010712132492600_bib54","doi-asserted-by":"crossref","first-page":"109","DOI":"10.1586\/14737159.9.2.109","article-title":"Spermatogonial stem cell biomarkers: improved outcomes of spermatogonial transplantation in male fertility restoration?","volume":"9","author":"Yeh","year":"2009","journal-title":"Expert Review of Molecular Diagnostics"},{"key":"2026010712132492600_bib55","doi-asserted-by":"crossref","first-page":"44","DOI":"10.1093\/humrep\/der357","article-title":"The efficiency of male fertility restoration is dependent on the recovery kinetics of spermatogonial stem cells after cytotoxic treatment with busulfan in mice","volume":"27","author":"Zohni","year":"2012","journal-title":"Human Reproduction"}],"container-title":["Reproduction"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/rep.bioscientifica.com\/view\/journals\/rep\/153\/6\/725.xml","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/academic.oup.com\/reproduction\/article-pdf\/153\/6\/725\/64120852\/725.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/academic.oup.com\/reproduction\/article-pdf\/153\/6\/725\/64120852\/725.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,1,7]],"date-time":"2026-01-07T17:14:20Z","timestamp":1767806060000},"score":1,"resource":{"primary":{"URL":"https:\/\/academic.oup.com\/reproduction\/article\/153\/6\/725\/8234980"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2017,6,1]]},"references-count":55,"journal-issue":{"issue":"6","published-print":{"date-parts":[[2017,6,1]]}},"URL":"https:\/\/doi.org\/10.1530\/rep-17-0005","relation":{},"ISSN":["1470-1626","1741-7899"],"issn-type":[{"value":"1470-1626","type":"print"},{"value":"1741-7899","type":"electronic"}],"subject":[],"published-other":{"date-parts":[[2017,6,1]]},"published":{"date-parts":[[2017,6,1]]}}}