Phosphorothioate-Modified AP613-1 Specifically Targets GPC3 when Used for Hepatocellular Carcinoma Cell Imaging
Glypican-3 (GPC3), the cellular membrane proteoglycan, has been established as a tumor biomarker for early diagnosis of hepatocellular carcinoma (HCC). GPC3 is highly expressed in more than 70% HCC tissues detected by antibody-based histopathological systems. Recently, aptamers, a short single-stran...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2018-12-01
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| Series: | Molecular Therapy: Nucleic Acids |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2162253118302579 |
| _version_ | 1828156370919096320 |
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| author | Lili Dong Hongxin Zhou Menglong Zhao Xinghui Gao Yang Liu Dongli Liu Wei Guo Hongwei Hu Qian Xie Jia Fan Jiang Lin Weizhong Wu |
| author_facet | Lili Dong Hongxin Zhou Menglong Zhao Xinghui Gao Yang Liu Dongli Liu Wei Guo Hongwei Hu Qian Xie Jia Fan Jiang Lin Weizhong Wu |
| author_sort | Lili Dong |
| collection | DOAJ |
| description | Glypican-3 (GPC3), the cellular membrane proteoglycan, has been established as a tumor biomarker for early diagnosis of hepatocellular carcinoma (HCC). GPC3 is highly expressed in more than 70% HCC tissues detected by antibody-based histopathological systems. Recently, aptamers, a short single-strand DNA or RNA generated from systematic evolution of ligands by exponential enrichment (SELEX), were reported as potential alternatives in tumor-targeted imaging and diagnosis. In this study, a total of 19 GPC3-bound aptamers were successfully screened by capillary electrophoresis (CE)-SELEX technology. After truncated, AP613-1 was confirmed to specifically target GPC3 with a dissociation constant (KD) of 59.85 nM. When modified with a phosphorothioate linkage, APS613-1 targeted GPC3 with a KD of 15.48 nM and could be used as a specific probe in living Huh7 and PLC/PRF/5 imaging, GPC3-positive cell lines, but not in L02 or A549, two GPC3-negative cell lines. More importantly, Alexa Fluor 750-conjugated APS613-1 could be used as a fluorescent probe to subcutaneous HCC imaging in xenograft nude mice. Our results indicated that modified AP613-1, especially APS613-1, was a potential agent in GPC3-positive tumor imaging for HCC early diagnosis. Keywords: GPC3, CE-SELEX, ssDNA aptamer, flow cytometry, cell imaging |
| first_indexed | 2024-04-11T23:10:49Z |
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| id | doaj.art-57b65506b9114f0996efe70de65c7a68 |
| institution | Directory Open Access Journal |
| issn | 2162-2531 |
| language | English |
| last_indexed | 2024-04-11T23:10:49Z |
| publishDate | 2018-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Therapy: Nucleic Acids |
| spelling | doaj.art-57b65506b9114f0996efe70de65c7a682022-12-22T03:57:51ZengElsevierMolecular Therapy: Nucleic Acids2162-25312018-12-0113376386Phosphorothioate-Modified AP613-1 Specifically Targets GPC3 when Used for Hepatocellular Carcinoma Cell ImagingLili Dong0Hongxin Zhou1Menglong Zhao2Xinghui Gao3Yang Liu4Dongli Liu5Wei Guo6Hongwei Hu7Qian Xie8Jia Fan9Jiang Lin10Weizhong Wu11Liver Cancer Institute, Zhongshan Hospital, Fudan University, Key Laboratory of Carcinogenesis and Cancer Invasion, Ministry of Education, Shanghai 200032, ChinaLiver Cancer Institute, Zhongshan Hospital, Fudan University, Key Laboratory of Carcinogenesis and Cancer Invasion, Ministry of Education, Shanghai 200032, ChinaDepartment of Radiology and Shanghai Institute of Medical Imaging, Zhongshan Hospital, Fudan University, Shanghai 200032, ChinaDepartment of Laboratory Medicine, Zhongshan Hospital, Fudan University, Shanghai 200032, ChinaLiver Cancer Institute, Zhongshan Hospital, Fudan University, Key Laboratory of Carcinogenesis and Cancer Invasion, Ministry of Education, Shanghai 200032, ChinaLiver Cancer Institute, Zhongshan Hospital, Fudan University, Key Laboratory of Carcinogenesis and Cancer Invasion, Ministry of Education, Shanghai 200032, China; Department of Radiation Oncology, Shanghai General Hospital, Shanghai Jiaotong University, Shanghai 200080, ChinaDepartment of Laboratory Medicine, Zhongshan Hospital, Fudan University, Shanghai 200032, ChinaShanghai Aijin Biochemical Science & Technology Co. Ltd., Shanghai 200336, ChinaCenter of Excellence in Inflammation, Infectious Disease and Immunity, James H. Quillen College of Medicine, East Tennessee State University, Johnson City, TN 37614, USALiver Cancer Institute, Zhongshan Hospital, Fudan University, Key Laboratory of Carcinogenesis and Cancer Invasion, Ministry of Education, Shanghai 200032, China; Institute of Biomedical Sciences, Fudan University, Shanghai 200032, ChinaDepartment of Radiology and Shanghai Institute of Medical Imaging, Zhongshan Hospital, Fudan University, Shanghai 200032, China; Institute of Functional and Molecular Medical Imaging, Fudan University, Shanghai 200040, China; Corresponding author: Jiang Lin, Liver Cancer Institute, Zhongshan Hospital, Fudan University, Key Laboratory of Carcinogenesis and Cancer Invasion, Ministry of Education, Fenglin Road 180, Shanghai 200032, China.Liver Cancer Institute, Zhongshan Hospital, Fudan University, Key Laboratory of Carcinogenesis and Cancer Invasion, Ministry of Education, Shanghai 200032, China; Corresponding author: Weizhong Wu, Liver Cancer Institute, Zhongshan Hospital, Fudan University, Key Laboratory of Carcinogenesis and Cancer Invasion, Ministry of Education, Fenglin Road 180, Shanghai 200032, China.Glypican-3 (GPC3), the cellular membrane proteoglycan, has been established as a tumor biomarker for early diagnosis of hepatocellular carcinoma (HCC). GPC3 is highly expressed in more than 70% HCC tissues detected by antibody-based histopathological systems. Recently, aptamers, a short single-strand DNA or RNA generated from systematic evolution of ligands by exponential enrichment (SELEX), were reported as potential alternatives in tumor-targeted imaging and diagnosis. In this study, a total of 19 GPC3-bound aptamers were successfully screened by capillary electrophoresis (CE)-SELEX technology. After truncated, AP613-1 was confirmed to specifically target GPC3 with a dissociation constant (KD) of 59.85 nM. When modified with a phosphorothioate linkage, APS613-1 targeted GPC3 with a KD of 15.48 nM and could be used as a specific probe in living Huh7 and PLC/PRF/5 imaging, GPC3-positive cell lines, but not in L02 or A549, two GPC3-negative cell lines. More importantly, Alexa Fluor 750-conjugated APS613-1 could be used as a fluorescent probe to subcutaneous HCC imaging in xenograft nude mice. Our results indicated that modified AP613-1, especially APS613-1, was a potential agent in GPC3-positive tumor imaging for HCC early diagnosis. Keywords: GPC3, CE-SELEX, ssDNA aptamer, flow cytometry, cell imaginghttp://www.sciencedirect.com/science/article/pii/S2162253118302579 |
| spellingShingle | Lili Dong Hongxin Zhou Menglong Zhao Xinghui Gao Yang Liu Dongli Liu Wei Guo Hongwei Hu Qian Xie Jia Fan Jiang Lin Weizhong Wu Phosphorothioate-Modified AP613-1 Specifically Targets GPC3 when Used for Hepatocellular Carcinoma Cell Imaging Molecular Therapy: Nucleic Acids |
| title | Phosphorothioate-Modified AP613-1 Specifically Targets GPC3 when Used for Hepatocellular Carcinoma Cell Imaging |
| title_full | Phosphorothioate-Modified AP613-1 Specifically Targets GPC3 when Used for Hepatocellular Carcinoma Cell Imaging |
| title_fullStr | Phosphorothioate-Modified AP613-1 Specifically Targets GPC3 when Used for Hepatocellular Carcinoma Cell Imaging |
| title_full_unstemmed | Phosphorothioate-Modified AP613-1 Specifically Targets GPC3 when Used for Hepatocellular Carcinoma Cell Imaging |
| title_short | Phosphorothioate-Modified AP613-1 Specifically Targets GPC3 when Used for Hepatocellular Carcinoma Cell Imaging |
| title_sort | phosphorothioate modified ap613 1 specifically targets gpc3 when used for hepatocellular carcinoma cell imaging |
| url | http://www.sciencedirect.com/science/article/pii/S2162253118302579 |
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