- Email: [email protected]
MDR1) IN DRUG-INDUCED LIVER INJURY
POSTERS vitro after the cells were treated with Ad.Luc, Ad.IL-24, Ad.Bax or Ad.IL-24-Bax. The mechanism involved was also explored. A model using nude mice with liver tumors was used to evaluate the adenovirus’ effects on tumor volume and cell apoptosis in vivo. Results: Ad.IL-24-Bax selectively suppressed growth of hepatocellular carcinoma cells but had little influence on the growth of normal hepatocytes in vitro. Ad.IL-24-Bax induced apoptosis in the hepatocellular carcinoma cells but not in normal hepatocytes. The Ad.IL-24-Bax had an enhanced ability to induce apoptosis and inhibit hepatocellular carcinoma cell growth when compared to Ad.IL-24 or Ad.Bax alone. The mechanism of this response may include the effect of the 10HRE/VEGF385 promoter and the synergistic effect of IL-24 and Bax. Ad.IL-24-Bax also suppressed tumor growth in nude mice and induced apoptosis. Conclusion: Ad.IL-24-Bax may be a useful tool for gene therapy in the treatment of hepatic cancer in clinical applications. 684 EFFECTS OF CHEMICAL MOIETIES AND POLYMORPHISMS IN THE HUMAN ABC TRANSPORTER ABCB1 (P-GLYCOPROTEIN/MDR1) IN DRUG-INDUCED LIVER INJURY 1
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E. Ulzurrun , E. Crespo , F. Ruiz-Cabello , C. Stephens , Y. Borraz , 5 M. Robles4 , M.R. Cabello1 , M.C. Fernandez ´ , G. Pelaez6 , 7 M. Romero-Gomez ´ , J.M. Navarro8 , R. Planas9 , J.F. Salmeron ´ 10 , 11 12 1 4 1 C. Guarner , A. Castiella , M.I. Lucena , R.J. Andrade . Servicio de Farmacolog´ıa Cl´ınica, Hospital Virgen de la Victoria, Facultad de Medicina, Universidad de M´ alaga. Centro de Investigaci´ on Biom´edica en Red de Enfermedades Hep´ aticas y Digestivas CIBERehd, M´ alaga, 2 Departamento de Farmacolog´ıa, Facultad de Farmacia, Universidad de Granada. Centro de Investigaci´ on Biom´edica en Red de Enfermedades Hep´ aticas y Digestivas CIBERehd, 3 Servicio de An´ alisis Cl´ınicos, Laboratorio de Inmunolog´ıa, Hospital Virgen de las Nieves, Granada, Granada, 4 Unidad de Hepatolog´ıa, Hospital Virgen de la Victoria, Facultad de Medicina. Universidad de M´ alaga. Centro de Investigaci´ on Biom´edica en Red de Enfermedades Hep´ aticas y Digestivas CIBERehd, M´ alaga, 5 Servicio de Farmacolog´ıa Cl´ınica, Hospital Torrec´ ardenas, Almer´ıa, 6 Unidad de Hepatolog´ıa, Hospital Torrec´ ardenas, Almer´ıa, 7 Unidad de Hepatolog´ıa, Hospital Nuestra Se˜ nora de Valme, Sevilla, Sevilla, 8 Unidad de Hepatolog´ıa, Hospital Costa del Sol, Marbella, 9 Unidad de Hepatolog´ıa, Hospital German Trias i Pujol. Centro de Investigaci´ on Biom´edica en Red de Enfermedades Hep´ aticas y Digestivas CIBERehd, Badalona, 10 Unidad de Hepatolog´ıa, Hospital Cl´ınico San Cecilio. Centro de Investigaci´ on Biom´edica en Red de Enfermedades Hep´ aticas y Digestivas CIBERehd, Granada, 11 Unidad de Hepatolog´ıa, Hospital Sant Pau. Centro de Investigaci´ on Biom´edica en Red de Enfermedades Hep´ aticas y Digestivas CIBERehd, Barcelona, 12 Unidad de Hepatolog´ıa, Hospital Mendaro, Guip´ uzcoa, Spain E-mail: [email protected]
Results: The genotype distribution for ABCB1 2677 G>T,A and 3435C>T polymorphisms did not show any significant differences when compared with controls, 2677TT (Pc = 1.000) and 3435TT (Pc = 0.946) respectively, neither for 2677TT-3435TT genotype (Pc = 1.000). The same result was observed for haplotype combinations (2677T-3435T; Pc = 1.000). Presence of ring-linking groups containing one C atom (M132) in the causative drug in patients with the ABCB1 2677T allele was associated with DILI development (Pc = 0.038; OR = 2.1), meanwhile the 2677G allele correlated with a protective effect (Pc = 0.026; OR = 0.5). Conclusions: Carriers of the ABCB1 2677T allele are at increased risk of developing DILI when exposed to drugs with ring-linking groups containing one C atom moieties. This study highlights the need to combine chemical structure information related to functional responses with genetic background, to more accurately predict individual susceptibility to DILI. Funding: Partially supported by Spanish Medicine Agency, grants SAS PI-00082/07 and FIS PS 09/01384. CIBERehd is funded by Instituto de Salud Carlos III. 685 L-CARBOCISTEINE REDUCED LIVER DAMAGE INDUCED BY OXALIPLATIN B. Yu1,2 . 1 Department of Pharmacy, Fudan University Shanghai Cancer Center, 2 Department of Oncology, Shanghai Medical College, Fudan University, Shanghai, China E-mail: [email protected] Background and Aims: Oxaliplatin (OX), a third-generation platinum drug used for the treatment of colorectal, lung, breast and ovarian cancers, could cause hepatic injury featured by oxidative damage and apoptosis. On the other hand, L-carbocisteine (CC) may inhibit cell damage induced by H2 O2 through the activation of Akt phosphorylation. The purpose of this investigation was to confirm whether the evaluation of the liver damage caused by OX using CC was practicable. Methods: Levels of ALT, ROS, MDA, CAT and GSH-PX were measured to identify liver function. HE and TUNEL assay was used to evaluate cell apoptosis in liver. Western blot was used to analyze signal cascade involved. Results: CC could inhibit OX induced significant apoptosis of hepatocytes by blockade of caspase 3 activation as well as upregulation of pAkt. In vivo tests showed anti-oxidative effects of CC against OX induced up-regulation of ROS and MDA and downregulation of CAT and GSH-PX. At the same time, the anti-tumor effect of OX remained unaltered. Conclusions: L-carbocisteine could reduce oxaliplatin induced liver damage without affecting anti-tumor effects of oxaliplatin.
Background and Aims: P-glycoprotein (P-gp) is a member of the adenosine triphosphate (ATP)-binding cassette family and is encoded by the human ABCB1 gene, also called MDR1 (multidrug resistance). P-gp is found in the canalicular surface of hepatocytes excreting drugs directly into the bile. We aimed to analyze whether characterised DILI patients had a higher prevalence of a variant genotype at positions 3435 in exon 26 and 2677 in exon 21 of ABCB1 and possible interactions with certain chemical moieties in the culprit drug structure that might enhance the risk of DILI. Methods: Genotyping of ABCB1 3435C>T (synonymous) and 2677G>T,A (A893S, T) was carried out in 157 Spanish DILI patients, assessed with the CIOMS scale, and 175 Spanish bone-marrow donors serving as the control group. Genotyping of 3435C>T and 2677G>T, A was analysed using the TaqMan 5 allelic discrimination assay and direct sequencing after, respectively. The presence of Chemical Fragmentation Codes (CFC) contributing to ATPase activity of ABCB1 as described by Sakurai et al (Biochemistry 2007), were analyzed for each individual drug causing DILI. Journal of Hepatology 2011 vol. 54 | S209–S361
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E. Ulzurrun , E. Crespo , F. Ruiz-Cabello , C. Stephens , Y. Borraz , 5 M. Robles4 , M.R. Cabello1 , M.C. Fernandez ´ , G. Pelaez6 , 7 M. Romero-Gomez ´ , J.M. Navarro8 , R. Planas9 , J.F. Salmeron ´ 10 , 11 12 1 4 1 C. Guarner , A. Castiella , M.I. Lucena , R.J. Andrade . Servicio de Farmacolog´ıa Cl´ınica, Hospital Virgen de la Victoria, Facultad de Medicina, Universidad de M´ alaga. Centro de Investigaci´ on Biom´edica en Red de Enfermedades Hep´ aticas y Digestivas CIBERehd, M´ alaga, 2 Departamento de Farmacolog´ıa, Facultad de Farmacia, Universidad de Granada. Centro de Investigaci´ on Biom´edica en Red de Enfermedades Hep´ aticas y Digestivas CIBERehd, 3 Servicio de An´ alisis Cl´ınicos, Laboratorio de Inmunolog´ıa, Hospital Virgen de las Nieves, Granada, Granada, 4 Unidad de Hepatolog´ıa, Hospital Virgen de la Victoria, Facultad de Medicina. Universidad de M´ alaga. Centro de Investigaci´ on Biom´edica en Red de Enfermedades Hep´ aticas y Digestivas CIBERehd, M´ alaga, 5 Servicio de Farmacolog´ıa Cl´ınica, Hospital Torrec´ ardenas, Almer´ıa, 6 Unidad de Hepatolog´ıa, Hospital Torrec´ ardenas, Almer´ıa, 7 Unidad de Hepatolog´ıa, Hospital Nuestra Se˜ nora de Valme, Sevilla, Sevilla, 8 Unidad de Hepatolog´ıa, Hospital Costa del Sol, Marbella, 9 Unidad de Hepatolog´ıa, Hospital German Trias i Pujol. Centro de Investigaci´ on Biom´edica en Red de Enfermedades Hep´ aticas y Digestivas CIBERehd, Badalona, 10 Unidad de Hepatolog´ıa, Hospital Cl´ınico San Cecilio. Centro de Investigaci´ on Biom´edica en Red de Enfermedades Hep´ aticas y Digestivas CIBERehd, Granada, 11 Unidad de Hepatolog´ıa, Hospital Sant Pau. Centro de Investigaci´ on Biom´edica en Red de Enfermedades Hep´ aticas y Digestivas CIBERehd, Barcelona, 12 Unidad de Hepatolog´ıa, Hospital Mendaro, Guip´ uzcoa, Spain E-mail: [email protected]
Results: The genotype distribution for ABCB1 2677 G>T,A and 3435C>T polymorphisms did not show any significant differences when compared with controls, 2677TT (Pc = 1.000) and 3435TT (Pc = 0.946) respectively, neither for 2677TT-3435TT genotype (Pc = 1.000). The same result was observed for haplotype combinations (2677T-3435T; Pc = 1.000). Presence of ring-linking groups containing one C atom (M132) in the causative drug in patients with the ABCB1 2677T allele was associated with DILI development (Pc = 0.038; OR = 2.1), meanwhile the 2677G allele correlated with a protective effect (Pc = 0.026; OR = 0.5). Conclusions: Carriers of the ABCB1 2677T allele are at increased risk of developing DILI when exposed to drugs with ring-linking groups containing one C atom moieties. This study highlights the need to combine chemical structure information related to functional responses with genetic background, to more accurately predict individual susceptibility to DILI. Funding: Partially supported by Spanish Medicine Agency, grants SAS PI-00082/07 and FIS PS 09/01384. CIBERehd is funded by Instituto de Salud Carlos III. 685 L-CARBOCISTEINE REDUCED LIVER DAMAGE INDUCED BY OXALIPLATIN B. Yu1,2 . 1 Department of Pharmacy, Fudan University Shanghai Cancer Center, 2 Department of Oncology, Shanghai Medical College, Fudan University, Shanghai, China E-mail: [email protected] Background and Aims: Oxaliplatin (OX), a third-generation platinum drug used for the treatment of colorectal, lung, breast and ovarian cancers, could cause hepatic injury featured by oxidative damage and apoptosis. On the other hand, L-carbocisteine (CC) may inhibit cell damage induced by H2 O2 through the activation of Akt phosphorylation. The purpose of this investigation was to confirm whether the evaluation of the liver damage caused by OX using CC was practicable. Methods: Levels of ALT, ROS, MDA, CAT and GSH-PX were measured to identify liver function. HE and TUNEL assay was used to evaluate cell apoptosis in liver. Western blot was used to analyze signal cascade involved. Results: CC could inhibit OX induced significant apoptosis of hepatocytes by blockade of caspase 3 activation as well as upregulation of pAkt. In vivo tests showed anti-oxidative effects of CC against OX induced up-regulation of ROS and MDA and downregulation of CAT and GSH-PX. At the same time, the anti-tumor effect of OX remained unaltered. Conclusions: L-carbocisteine could reduce oxaliplatin induced liver damage without affecting anti-tumor effects of oxaliplatin.
Background and Aims: P-glycoprotein (P-gp) is a member of the adenosine triphosphate (ATP)-binding cassette family and is encoded by the human ABCB1 gene, also called MDR1 (multidrug resistance). P-gp is found in the canalicular surface of hepatocytes excreting drugs directly into the bile. We aimed to analyze whether characterised DILI patients had a higher prevalence of a variant genotype at positions 3435 in exon 26 and 2677 in exon 21 of ABCB1 and possible interactions with certain chemical moieties in the culprit drug structure that might enhance the risk of DILI. Methods: Genotyping of ABCB1 3435C>T (synonymous) and 2677G>T,A (A893S, T) was carried out in 157 Spanish DILI patients, assessed with the CIOMS scale, and 175 Spanish bone-marrow donors serving as the control group. Genotyping of 3435C>T and 2677G>T, A was analysed using the TaqMan 5 allelic discrimination assay and direct sequencing after, respectively. The presence of Chemical Fragmentation Codes (CFC) contributing to ATPase activity of ABCB1 as described by Sakurai et al (Biochemistry 2007), were analyzed for each individual drug causing DILI. Journal of Hepatology 2011 vol. 54 | S209–S361
S275