Establishing protocol for detecting methylation of Ubiquitin carboxyl terminal hydrolase 1 gene’s promoter in nasopharyngeal carcinoma
Background: The methylation of Ubiquitin Carboxyl Terminal Hydrolase 1 (UCHL1) gene’s promoter has been reported as the etiological factor of nasopharyngeal tumorigenesis. Purpose: This study is designed to establish a protocol for detecting methylation of UCHL1 gene’s promoter in nasopharyngeal car...
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| Format: | Article |
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HO CHI MINH CITY OPEN UNIVERSITY JOURNAL OF SCIENCE
2022-11-01
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| Series: | Ho Chi Minh City Open University Journal of Science - Engineering and Technology |
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| Online Access: | https://journalofscience.ou.edu.vn/index.php/tech-en/article/view/2542 |
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| author | Thieu Hong Hue Huynh Thi Ngoc Tuyen Nguyen Van Truong Dao Duy Tin Tran Minh Dang Khoa Le Huyen Ai Thuy Lao Duc Thuan |
| author_facet | Thieu Hong Hue Huynh Thi Ngoc Tuyen Nguyen Van Truong Dao Duy Tin Tran Minh Dang Khoa Le Huyen Ai Thuy Lao Duc Thuan |
| author_sort | Thieu Hong Hue |
| collection | DOAJ |
| description | Background: The methylation of Ubiquitin Carboxyl Terminal Hydrolase 1 (UCHL1) gene’s promoter has been reported as the etiological factor of nasopharyngeal tumorigenesis. Purpose: This study is designed to establish a protocol for detecting methylation of UCHL1 gene’s promoter in nasopharyngeal carcinoma (NPC) in a Vietnamese population. Materials and methods: 10 samples of NPC biopsy tissues and 10 samples of non-cancerous swabs were collected from the local hospital. Chloroform/Phenol method and Nested-MSP assays were established to detect methylation of a target gene. Results: The isolated DNA reached purity and high concentration which were confirmed by the method of absorbance measurement at 260nm and 280nm. Additionally, the Nested-MSP products of methylation or unmethylation were analyzed and visualized in the agarose gel with the band of 169bps and 210bps, respectively. By sequencing, it was confirmed that the two sets of primer could distinguish the status of methylation and unmethylation of UCHL1 gene’s promoter. Conclusion: Our data suggested that the current protocol could successfully identify the status of methylation and/or unmethylation of UCHL1 gene’s promoter. |
| first_indexed | 2024-04-11T06:31:43Z |
| format | Article |
| id | doaj.art-63f6360f4fd24d91b2935e61b1e94f37 |
| institution | Directory Open Access Journal |
| issn | 2734-9330 2734-9608 |
| language | English |
| last_indexed | 2024-04-11T06:31:43Z |
| publishDate | 2022-11-01 |
| publisher | HO CHI MINH CITY OPEN UNIVERSITY JOURNAL OF SCIENCE |
| record_format | Article |
| series | Ho Chi Minh City Open University Journal of Science - Engineering and Technology |
| spelling | doaj.art-63f6360f4fd24d91b2935e61b1e94f372022-12-22T04:40:01ZengHO CHI MINH CITY OPEN UNIVERSITY JOURNAL OF SCIENCEHo Chi Minh City Open University Journal of Science - Engineering and Technology2734-93302734-96082022-11-01122344110.46223/HCMCOUJS.tech.en.12.2.2542.20221782Establishing protocol for detecting methylation of Ubiquitin carboxyl terminal hydrolase 1 gene’s promoter in nasopharyngeal carcinomaThieu Hong Hue0Huynh Thi Ngoc Tuyen1Nguyen Van Truong2Dao Duy Tin3Tran Minh Dang Khoa4Le Huyen Ai Thuy5Lao Duc Thuan6Ho Chi Minh City Open University, Ho Chi Minh CityHo Chi Minh City Open University, Ho Chi Minh CityHo Chi Minh City Open University, Ho Chi Minh CityHo Chi Minh City Open University, Ho Chi Minh CityHo Chi Minh City Open University, Ho Chi Minh CityHo Chi Minh City Open University, Ho Chi Minh CityHo Chi Minh City Open University, Ho Chi Minh CityBackground: The methylation of Ubiquitin Carboxyl Terminal Hydrolase 1 (UCHL1) gene’s promoter has been reported as the etiological factor of nasopharyngeal tumorigenesis. Purpose: This study is designed to establish a protocol for detecting methylation of UCHL1 gene’s promoter in nasopharyngeal carcinoma (NPC) in a Vietnamese population. Materials and methods: 10 samples of NPC biopsy tissues and 10 samples of non-cancerous swabs were collected from the local hospital. Chloroform/Phenol method and Nested-MSP assays were established to detect methylation of a target gene. Results: The isolated DNA reached purity and high concentration which were confirmed by the method of absorbance measurement at 260nm and 280nm. Additionally, the Nested-MSP products of methylation or unmethylation were analyzed and visualized in the agarose gel with the band of 169bps and 210bps, respectively. By sequencing, it was confirmed that the two sets of primer could distinguish the status of methylation and unmethylation of UCHL1 gene’s promoter. Conclusion: Our data suggested that the current protocol could successfully identify the status of methylation and/or unmethylation of UCHL1 gene’s promoter.https://journalofscience.ou.edu.vn/index.php/tech-en/article/view/2542epigeneticsmethylationnested-mspubiquitin carboxyl terminal hydrolase 1unmethylation |
| spellingShingle | Thieu Hong Hue Huynh Thi Ngoc Tuyen Nguyen Van Truong Dao Duy Tin Tran Minh Dang Khoa Le Huyen Ai Thuy Lao Duc Thuan Establishing protocol for detecting methylation of Ubiquitin carboxyl terminal hydrolase 1 gene’s promoter in nasopharyngeal carcinoma Ho Chi Minh City Open University Journal of Science - Engineering and Technology epigenetics methylation nested-msp ubiquitin carboxyl terminal hydrolase 1 unmethylation |
| title | Establishing protocol for detecting methylation of Ubiquitin carboxyl terminal hydrolase 1 gene’s promoter in nasopharyngeal carcinoma |
| title_full | Establishing protocol for detecting methylation of Ubiquitin carboxyl terminal hydrolase 1 gene’s promoter in nasopharyngeal carcinoma |
| title_fullStr | Establishing protocol for detecting methylation of Ubiquitin carboxyl terminal hydrolase 1 gene’s promoter in nasopharyngeal carcinoma |
| title_full_unstemmed | Establishing protocol for detecting methylation of Ubiquitin carboxyl terminal hydrolase 1 gene’s promoter in nasopharyngeal carcinoma |
| title_short | Establishing protocol for detecting methylation of Ubiquitin carboxyl terminal hydrolase 1 gene’s promoter in nasopharyngeal carcinoma |
| title_sort | establishing protocol for detecting methylation of ubiquitin carboxyl terminal hydrolase 1 gene s promoter in nasopharyngeal carcinoma |
| topic | epigenetics methylation nested-msp ubiquitin carboxyl terminal hydrolase 1 unmethylation |
| url | https://journalofscience.ou.edu.vn/index.php/tech-en/article/view/2542 |
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