MiR-20a-5p suppresses tumor proliferation by targeting autophagy-related gene 7 in neuroblastoma
Abstract Background Neuroblastoma (NB) is the most common malignant tumor originating from the extracranial sympathetic nervous system in children. The molecular mechanisms underlying this disease are complex, and not completely understood. Methods Quantitative real-time PCR (qRT-PCR) was applied to...
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| Format: | Article |
| Language: | English |
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BMC
2018-01-01
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| Series: | Cancer Cell International |
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| Online Access: | http://link.springer.com/article/10.1186/s12935-017-0499-2 |
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doaj-0db5fdd35e2446f6a6688967c33a250d |
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Article |
| collection |
DOAJ |
| language |
English |
| format |
Article |
| sources |
DOAJ |
| author |
Yongbo Yu Jie Zhang Yaqiong Jin Yeran Yang Jin Shi Feng Chen Shujing Han Ping Chu Jie Lu Huanmin Wang Yongli Guo Xin Ni |
| spellingShingle |
Yongbo Yu Jie Zhang Yaqiong Jin Yeran Yang Jin Shi Feng Chen Shujing Han Ping Chu Jie Lu Huanmin Wang Yongli Guo Xin Ni MiR-20a-5p suppresses tumor proliferation by targeting autophagy-related gene 7 in neuroblastoma Cancer Cell International Neuroblastoma miR-20a-5p Autophagy Autophagy-related gene 7 Pediatrics |
| author_facet |
Yongbo Yu Jie Zhang Yaqiong Jin Yeran Yang Jin Shi Feng Chen Shujing Han Ping Chu Jie Lu Huanmin Wang Yongli Guo Xin Ni |
| author_sort |
Yongbo Yu |
| title |
MiR-20a-5p suppresses tumor proliferation by targeting autophagy-related gene 7 in neuroblastoma |
| title_short |
MiR-20a-5p suppresses tumor proliferation by targeting autophagy-related gene 7 in neuroblastoma |
| title_full |
MiR-20a-5p suppresses tumor proliferation by targeting autophagy-related gene 7 in neuroblastoma |
| title_fullStr |
MiR-20a-5p suppresses tumor proliferation by targeting autophagy-related gene 7 in neuroblastoma |
| title_full_unstemmed |
MiR-20a-5p suppresses tumor proliferation by targeting autophagy-related gene 7 in neuroblastoma |
| title_sort |
mir-20a-5p suppresses tumor proliferation by targeting autophagy-related gene 7 in neuroblastoma |
| publisher |
BMC |
| series |
Cancer Cell International |
| issn |
1475-2867 |
| publishDate |
2018-01-01 |
| description |
Abstract Background Neuroblastoma (NB) is the most common malignant tumor originating from the extracranial sympathetic nervous system in children. The molecular mechanisms underlying this disease are complex, and not completely understood. Methods Quantitative real-time PCR (qRT-PCR) was applied to quantify the expression of miR-20a-5p and its target gene ATG7 in clinical NB tissues. The biological function of miR-20a-5p and ATG7 in SH-SY5Y cells was investigated through in vitro studies (Real-Time cell kinetic analyzer, colony formation assay, caspase-Glo 3/7 assay and western blotting). The luciferase reporter assay was conducted to verify the biological relationship between miR-20a-5p and ATG7. Results Here we found that miR-20a-5p expression was significantly downregulated whereas its target autophagy-related gene 7 (ATG7) was increased along with clinical staging of NB progression. Correlation analysis showed that miR-20a-5p had a negative correlation trend with ATG7. In SH-SY5Y cells, forced expression of miR-20a-5p suppressed ATG7 expression, autophagy initiation and cellular proliferation while promoted apoptosis, suggesting a potential association between miR-20a-5p and ATG7. Further bioinformatic target prediction combined with protein expression and luciferase reporter assay verified that miR-20a-5p inhibited ATG7 by directly binding to its 3′-UTR, confirming the involvement of miR-20a-5p in the regulation of ATG7 in NB. Conclusions These results clarified that miR-20a-5p inhibited cell proliferation and promoted apoptosis through negative regulation of ATG7 and thus autophagy suppression in SH-SY5Y cells. Therefore, defining the context-specific roles of autophagy in NB and regulatory mechanisms involved will be critical for developing autophagy-targeted therapeutics against NB. Both miR-20a-5p and ATG7 would be potential therapeutic targets for future NB treatment. |
| topic |
Neuroblastoma miR-20a-5p Autophagy Autophagy-related gene 7 Pediatrics |
| url |
http://link.springer.com/article/10.1186/s12935-017-0499-2 |
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doaj-0db5fdd35e2446f6a6688967c33a250d2020-11-24T23:05:58ZengBMCCancer Cell International1475-28672018-01-0118111010.1186/s12935-017-0499-2MiR-20a-5p suppresses tumor proliferation by targeting autophagy-related gene 7 in neuroblastomaYongbo Yu0Jie Zhang1Yaqiong Jin2Yeran Yang3Jin Shi4Feng Chen5Shujing Han6Ping Chu7Jie Lu8Huanmin Wang9Yongli Guo10Xin Ni11Beijing Key Laboratory for Pediatric Diseases of Otolaryngology, Head and Neck Surgery, MOE Key Laboratory of Major Diseases in Children, Beijing Pediatric Research Institute, Beijing Children’s Hospital, Capital Medical University, National Center for Children’s Health (NCCH)Beijing Key Laboratory for Pediatric Diseases of Otolaryngology, Head and Neck Surgery, MOE Key Laboratory of Major Diseases in Children, Beijing Pediatric Research Institute, Beijing Children’s Hospital, Capital Medical University, National Center for Children’s Health (NCCH)Beijing Key Laboratory for Pediatric Diseases of Otolaryngology, Head and Neck Surgery, MOE Key Laboratory of Major Diseases in Children, Beijing Pediatric Research Institute, Beijing Children’s Hospital, Capital Medical University, National Center for Children’s Health (NCCH)Beijing Key Laboratory for Pediatric Diseases of Otolaryngology, Head and Neck Surgery, MOE Key Laboratory of Major Diseases in Children, Beijing Pediatric Research Institute, Beijing Children’s Hospital, Capital Medical University, National Center for Children’s Health (NCCH)Beijing Key Laboratory for Pediatric Diseases of Otolaryngology, Head and Neck Surgery, MOE Key Laboratory of Major Diseases in Children, Beijing Pediatric Research Institute, Beijing Children’s Hospital, Capital Medical University, National Center for Children’s Health (NCCH)Beijing Key Laboratory for Pediatric Diseases of Otolaryngology, Head and Neck Surgery, MOE Key Laboratory of Major Diseases in Children, Beijing Pediatric Research Institute, Beijing Children’s Hospital, Capital Medical University, National Center for Children’s Health (NCCH)Beijing Key Laboratory for Pediatric Diseases of Otolaryngology, Head and Neck Surgery, MOE Key Laboratory of Major Diseases in Children, Beijing Pediatric Research Institute, Beijing Children’s Hospital, Capital Medical University, National Center for Children’s Health (NCCH)Beijing Key Laboratory for Pediatric Diseases of Otolaryngology, Head and Neck Surgery, MOE Key Laboratory of Major Diseases in Children, Beijing Pediatric Research Institute, Beijing Children’s Hospital, Capital Medical University, National Center for Children’s Health (NCCH)Beijing Key Laboratory for Pediatric Diseases of Otolaryngology, Head and Neck Surgery, MOE Key Laboratory of Major Diseases in Children, Beijing Pediatric Research Institute, Beijing Children’s Hospital, Capital Medical University, National Center for Children’s Health (NCCH)Department of Surgical Oncology, Beijing Children’s Hospital, Capital Medical University, National Center for Children’s Health (NCCH)Beijing Key Laboratory for Pediatric Diseases of Otolaryngology, Head and Neck Surgery, MOE Key Laboratory of Major Diseases in Children, Beijing Pediatric Research Institute, Beijing Children’s Hospital, Capital Medical University, National Center for Children’s Health (NCCH)Beijing Key Laboratory for Pediatric Diseases of Otolaryngology, Head and Neck Surgery, MOE Key Laboratory of Major Diseases in Children, Beijing Pediatric Research Institute, Beijing Children’s Hospital, Capital Medical University, National Center for Children’s Health (NCCH)Abstract Background Neuroblastoma (NB) is the most common malignant tumor originating from the extracranial sympathetic nervous system in children. The molecular mechanisms underlying this disease are complex, and not completely understood. Methods Quantitative real-time PCR (qRT-PCR) was applied to quantify the expression of miR-20a-5p and its target gene ATG7 in clinical NB tissues. The biological function of miR-20a-5p and ATG7 in SH-SY5Y cells was investigated through in vitro studies (Real-Time cell kinetic analyzer, colony formation assay, caspase-Glo 3/7 assay and western blotting). The luciferase reporter assay was conducted to verify the biological relationship between miR-20a-5p and ATG7. Results Here we found that miR-20a-5p expression was significantly downregulated whereas its target autophagy-related gene 7 (ATG7) was increased along with clinical staging of NB progression. Correlation analysis showed that miR-20a-5p had a negative correlation trend with ATG7. In SH-SY5Y cells, forced expression of miR-20a-5p suppressed ATG7 expression, autophagy initiation and cellular proliferation while promoted apoptosis, suggesting a potential association between miR-20a-5p and ATG7. Further bioinformatic target prediction combined with protein expression and luciferase reporter assay verified that miR-20a-5p inhibited ATG7 by directly binding to its 3′-UTR, confirming the involvement of miR-20a-5p in the regulation of ATG7 in NB. Conclusions These results clarified that miR-20a-5p inhibited cell proliferation and promoted apoptosis through negative regulation of ATG7 and thus autophagy suppression in SH-SY5Y cells. Therefore, defining the context-specific roles of autophagy in NB and regulatory mechanisms involved will be critical for developing autophagy-targeted therapeutics against NB. Both miR-20a-5p and ATG7 would be potential therapeutic targets for future NB treatment.http://link.springer.com/article/10.1186/s12935-017-0499-2NeuroblastomamiR-20a-5pAutophagyAutophagy-related gene 7Pediatrics |