BACKGROUND: To investigate mitochondrial (mt) toxicity of antiretroviral drugs further, we developed a novel real-time PCR-based assay for the quantification of mtRNA. We analysed the effects of stavudine (d4T), didanosine (ddl) and zidovudine (AZT) on the production of mtRNAs in different human cell lines and compared the production with the amount of mtDNA present in the same cells. MATERIALS AND METHODS: HUT78, CEM and U937 cells were exposed to different nucleoside reverse transcriptase inhibitors (NRTIs) for 7 days. Thereafter, nucleic acids were isolated and Taqman-based real-time PCR was used to quantify mtDNA and three different mtRNAs (ND1, CYTB and ND6 gene products). RESULTS: Different amounts of mtRNAs exist in different cell lines. When mtRNA was measured in cells exposed to an NRTI, a marked decrease was observed in cells treated with d4T, but not with ddl or AZT. Changes in mtRNA production did not always correspond to modifications in mtDNA content: 1 microM d4T significantly changed mtRNA but not mtDNA content. CONCLUSIONS: d4T, but not ddl or AZT, significantly alters mtRNA quantity and quality. The method we have developed can reveal changes that are not observed by measuring mtDNA content only, and can be used for ex vivo studies on drug toxicity.
BACKGROUND: To investigate mitochondrial (mt) toxicity of antiretroviral drugs further, we developed a novel real-time PCR-based assay for the quantification of mtRNA. We analysed the effects of stavudine (d4T), didanosine (ddl) and zidovudine (AZT) on the production of mtRNAs in different human cell lines and compared the production with the amount of mtDNA present in the same cells. MATERIALS AND METHODS: HUT78, CEM and U937 cells were exposed to different nucleoside reverse transcriptase inhibitors (NRTIs) for 7 days. Thereafter, nucleic acids were isolated and Taqman-based real-time PCR was used to quantify mtDNA and three different mtRNAs (ND1, CYTB and ND6 gene products). RESULTS: Different amounts of mtRNAs exist in different cell lines. When mtRNA was measured in cells exposed to an NRTI, a marked decrease was observed in cells treated with d4T, but not with ddl or AZT. Changes in mtRNA production did not always correspond to modifications in mtDNA content: 1 microM d4T significantly changed mtRNA but not mtDNA content. CONCLUSIONS: d4T, but not ddl or AZT, significantly alters mtRNA quantity and quality. The method we have developed can reveal changes that are not observed by measuring mtDNA content only, and can be used for ex vivo studies on drug toxicity.
Authors: Hélène C F Côté; Mariana Gerschenson; Ulrich A Walker; Oscar Miro; Gloria Garrabou; Emma Hammond; Joan Villarroya; Marta Giralt; Francesc Villarroya; Paola Cinque; Elena Garcia-Arumi; Antonio L Andreu; Marcello Pinti; Andrea Cossarizza Journal: Mitochondrion Date: 2011-02-22 Impact factor: 4.160
Authors: M Nasi; S De Biasi; L Gibellini; E Bianchini; S Pecorini; V Bacca; G Guaraldi; C Mussini; M Pinti; A Cossarizza Journal: Clin Exp Immunol Date: 2016-08-09 Impact factor: 4.330
Authors: Ankit Shah; Mohitkumar R Gangwani; Nitish S Chaudhari; Alexy Glazyrin; Hari K Bhat; Anil Kumar Journal: Neurotox Res Date: 2016-06-30 Impact factor: 3.911
Authors: Akihiko Saitoh; Richard H Haas; Robert K Naviaux; Neurita G Salva; Justine K Wong; Stephen A Spector Journal: Antimicrob Agents Chemother Date: 2008-06-09 Impact factor: 5.191
Authors: Grace A McComsey; Daniel E Libutti; MaryAnn O'Riordan; Jessica M Shelton; Norma Storer; Jason Ganz; John Jasper; Danielle Harrill; Mariana Gerschenson Journal: Antivir Ther Date: 2008
Authors: John R Koethe; Wyatt McDonnell; Arion Kennedy; Chike O Abana; Mark Pilkinton; Ian Setliff; Ivelin Georgiev; Louise Barnett; Cindy C Hager; Rita Smith; Spyros A Kalams; Alyssa Hasty; Simon Mallal Journal: J Acquir Immune Defic Syndr Date: 2018-02-01 Impact factor: 3.731