Caspase-Independent Pathway is Related to Nilotinib Cytotoxicity in Cultured Cardiomyocytes
Background/Aims: Cardiotoxicity is a predominant side-effect of nilotinib during chronic myeloid leukemia treatment. The underlying molecular mechanism remains unclear. The role of autophagy and mitochondrial signaling was investigated in nilotinib-treated cardiac H9C2 cells. Methods: Cytotoxicity w...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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Cell Physiol Biochem Press GmbH & Co KG
2017-08-01
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| Series: | Cellular Physiology and Biochemistry |
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| Online Access: | http://www.karger.com/Article/FullText/479993 |
| _version_ | 1818172841244229632 |
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| author | Qinghui Yang Chunhui Zhang Hong Wei Zenghui Meng Guangnan Li Yuanyuan Xu Yanjun Chen |
| author_facet | Qinghui Yang Chunhui Zhang Hong Wei Zenghui Meng Guangnan Li Yuanyuan Xu Yanjun Chen |
| author_sort | Qinghui Yang |
| collection | DOAJ |
| description | Background/Aims: Cardiotoxicity is a predominant side-effect of nilotinib during chronic myeloid leukemia treatment. The underlying molecular mechanism remains unclear. The role of autophagy and mitochondrial signaling was investigated in nilotinib-treated cardiac H9C2 cells. Methods: Cytotoxicity was assessed using Cell Death Detection kit. Immunoblot and immunofluorescence staining was performed, and cathepsin B and caspase3 activity was assessed in nilotinib-treated H9C2 cells with or without distinct pathway inhibitor or specific siRNA. Results: Nilotinib time- and dose-dependently induced H9C2 apoptosis, which was not completely prevented by the pan caspase inhibitor z-VAD-fmk. Following nilotinib treatment, mitochondrial membrane potential decreased significantly accompanied with remarkable morphological changes. Nuclear translocation of mitochondrial apoptosis inducing factor (AIF) and increased p53 was detected in nilotinib-treated cells. AIF knockdown prevented nilotinib-induced increase of p53 and apoptosis. Additionally, increased cathepsin B activity was detected, and inhibition of cathepsin B by CA-074Me prevented nilotinib-induced apoptosis and nuclear translocation of AIF. Moreover, increased Atg5 and transition of LC3-I to LC3-II was revealed following nilotinib treatment. Increased cathepsin B activity and apoptosis by nilotinib was significantly prohibited by specific autophagy inhibitor bafilomycin A and Atg5 knockdown. Conclusion: Our findings demonstrate that nilotinib increases autophagy and cathepsin B activity, leading to mitochondrial AIF release and nuclear translocation, which is responsible for p53 and apoptosis induction in H9C2 cells. |
| first_indexed | 2024-12-11T19:19:01Z |
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| institution | Directory Open Access Journal |
| issn | 1015-8987 1421-9778 |
| language | English |
| last_indexed | 2024-12-11T19:19:01Z |
| publishDate | 2017-08-01 |
| publisher | Cell Physiol Biochem Press GmbH & Co KG |
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| series | Cellular Physiology and Biochemistry |
| spelling | doaj.art-92d7e2c9a96b461f853d182fd50673ed2022-12-22T00:53:35ZengCell Physiol Biochem Press GmbH & Co KGCellular Physiology and Biochemistry1015-89871421-97782017-08-014262182219310.1159/000479993479993Caspase-Independent Pathway is Related to Nilotinib Cytotoxicity in Cultured CardiomyocytesQinghui YangChunhui ZhangHong WeiZenghui MengGuangnan LiYuanyuan XuYanjun ChenBackground/Aims: Cardiotoxicity is a predominant side-effect of nilotinib during chronic myeloid leukemia treatment. The underlying molecular mechanism remains unclear. The role of autophagy and mitochondrial signaling was investigated in nilotinib-treated cardiac H9C2 cells. Methods: Cytotoxicity was assessed using Cell Death Detection kit. Immunoblot and immunofluorescence staining was performed, and cathepsin B and caspase3 activity was assessed in nilotinib-treated H9C2 cells with or without distinct pathway inhibitor or specific siRNA. Results: Nilotinib time- and dose-dependently induced H9C2 apoptosis, which was not completely prevented by the pan caspase inhibitor z-VAD-fmk. Following nilotinib treatment, mitochondrial membrane potential decreased significantly accompanied with remarkable morphological changes. Nuclear translocation of mitochondrial apoptosis inducing factor (AIF) and increased p53 was detected in nilotinib-treated cells. AIF knockdown prevented nilotinib-induced increase of p53 and apoptosis. Additionally, increased cathepsin B activity was detected, and inhibition of cathepsin B by CA-074Me prevented nilotinib-induced apoptosis and nuclear translocation of AIF. Moreover, increased Atg5 and transition of LC3-I to LC3-II was revealed following nilotinib treatment. Increased cathepsin B activity and apoptosis by nilotinib was significantly prohibited by specific autophagy inhibitor bafilomycin A and Atg5 knockdown. Conclusion: Our findings demonstrate that nilotinib increases autophagy and cathepsin B activity, leading to mitochondrial AIF release and nuclear translocation, which is responsible for p53 and apoptosis induction in H9C2 cells.http://www.karger.com/Article/FullText/479993H9c2ApoptosisAtg5LC3AIFAutophagyCathepsin BMitochondria |
| spellingShingle | Qinghui Yang Chunhui Zhang Hong Wei Zenghui Meng Guangnan Li Yuanyuan Xu Yanjun Chen Caspase-Independent Pathway is Related to Nilotinib Cytotoxicity in Cultured Cardiomyocytes Cellular Physiology and Biochemistry H9c2 Apoptosis Atg5 LC3 AIF Autophagy Cathepsin B Mitochondria |
| title | Caspase-Independent Pathway is Related to Nilotinib Cytotoxicity in Cultured Cardiomyocytes |
| title_full | Caspase-Independent Pathway is Related to Nilotinib Cytotoxicity in Cultured Cardiomyocytes |
| title_fullStr | Caspase-Independent Pathway is Related to Nilotinib Cytotoxicity in Cultured Cardiomyocytes |
| title_full_unstemmed | Caspase-Independent Pathway is Related to Nilotinib Cytotoxicity in Cultured Cardiomyocytes |
| title_short | Caspase-Independent Pathway is Related to Nilotinib Cytotoxicity in Cultured Cardiomyocytes |
| title_sort | caspase independent pathway is related to nilotinib cytotoxicity in cultured cardiomyocytes |
| topic | H9c2 Apoptosis Atg5 LC3 AIF Autophagy Cathepsin B Mitochondria |
| url | http://www.karger.com/Article/FullText/479993 |
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