Identification and characterization of microRNA candidates that are upregulated by 5-aza-2’-deoxycytidine treatment in hepatocellular carcinoma cells.
MicroRNAs are small non-coding RNA of 19-20 nucleotides, capable of negatively regulating gene expression by targeting specific mRNA gene sequences, which causes mRNA degradation or inhibits protein translation. This study aims to investigate whether DNA hypermethylation may contribute to microRNA s...
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| Format: | Final Year Project (FYP) |
| Language: | English |
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2009
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| Online Access: | http://hdl.handle.net/10356/16345 |
| _version_ | 1811684066425044992 |
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| author | Leng, Charmaine Yong En. |
| author2 | School of Biological Sciences |
| author_facet | School of Biological Sciences Leng, Charmaine Yong En. |
| author_sort | Leng, Charmaine Yong En. |
| collection | NTU |
| description | MicroRNAs are small non-coding RNA of 19-20 nucleotides, capable of negatively regulating gene expression by targeting specific mRNA gene sequences, which causes mRNA degradation or inhibits protein translation. This study aims to investigate whether DNA hypermethylation may contribute to microRNA suppression, thereby modulating their target mRNA expression during the development of hepatocellular carcinoma (HCC). Preliminary microarray data revealed 19 up-regulated microRNAs in Hep3B cells treated with a DNA demethylation agent, 5-aza-2’-deoxycytidine (5-aza-dC), with hsa-miR-886-5p showing the greatest increase in expression level. Using TaqMan RT-PCR, my finding confirmed that hsa-miR-886-5p expression was consistently up-regulated after 5-aza-dC treatment in 3 HCC cell lines – Hep3B, HepG2 and PLC/PRF/5. More than half of the cancerous liver tissues from 17 HCC patients were observed to have down-regulated hsa-miR-886-5p expression as compared with their surrounding non-cancerous liver tissues. To determine putative mRNA targets of hsa-miR-886-5p, HCC cells were overexpressed ectopically with hsa-miR-886-5p and screened for differentially expressed genes using microarrays. 102 and 53 probe sets were consistently down-regulated and up-regulated respectively, in all 3 HCC cell lines. Real time RT-PCR was performed to validate the microarray results. These findings might provide new insights to the mechanism regulating hsa-miR-886-5p, and the network of genes targeted by hsa-miR-886-5p. |
| first_indexed | 2024-10-01T04:22:43Z |
| format | Final Year Project (FYP) |
| id | ntu-10356/16345 |
| institution | Nanyang Technological University |
| language | English |
| last_indexed | 2024-10-01T04:22:43Z |
| publishDate | 2009 |
| record_format | dspace |
| spelling | ntu-10356/163452023-02-28T18:05:21Z Identification and characterization of microRNA candidates that are upregulated by 5-aza-2’-deoxycytidine treatment in hepatocellular carcinoma cells. Leng, Charmaine Yong En. School of Biological Sciences Department of Clinical Research, Singapore General Hospital Tan, Michelle Guet Khim DRNTU::Science::Biological sciences::Genetics MicroRNAs are small non-coding RNA of 19-20 nucleotides, capable of negatively regulating gene expression by targeting specific mRNA gene sequences, which causes mRNA degradation or inhibits protein translation. This study aims to investigate whether DNA hypermethylation may contribute to microRNA suppression, thereby modulating their target mRNA expression during the development of hepatocellular carcinoma (HCC). Preliminary microarray data revealed 19 up-regulated microRNAs in Hep3B cells treated with a DNA demethylation agent, 5-aza-2’-deoxycytidine (5-aza-dC), with hsa-miR-886-5p showing the greatest increase in expression level. Using TaqMan RT-PCR, my finding confirmed that hsa-miR-886-5p expression was consistently up-regulated after 5-aza-dC treatment in 3 HCC cell lines – Hep3B, HepG2 and PLC/PRF/5. More than half of the cancerous liver tissues from 17 HCC patients were observed to have down-regulated hsa-miR-886-5p expression as compared with their surrounding non-cancerous liver tissues. To determine putative mRNA targets of hsa-miR-886-5p, HCC cells were overexpressed ectopically with hsa-miR-886-5p and screened for differentially expressed genes using microarrays. 102 and 53 probe sets were consistently down-regulated and up-regulated respectively, in all 3 HCC cell lines. Real time RT-PCR was performed to validate the microarray results. These findings might provide new insights to the mechanism regulating hsa-miR-886-5p, and the network of genes targeted by hsa-miR-886-5p. Bachelor of Science in Biological Sciences 2009-05-25T06:53:47Z 2009-05-25T06:53:47Z 2009 2009 Final Year Project (FYP) http://hdl.handle.net/10356/16345 en Nanyang Technological University 31 p. application/pdf |
| spellingShingle | DRNTU::Science::Biological sciences::Genetics Leng, Charmaine Yong En. Identification and characterization of microRNA candidates that are upregulated by 5-aza-2’-deoxycytidine treatment in hepatocellular carcinoma cells. |
| title | Identification and characterization of microRNA candidates that are upregulated by 5-aza-2’-deoxycytidine treatment in hepatocellular carcinoma cells. |
| title_full | Identification and characterization of microRNA candidates that are upregulated by 5-aza-2’-deoxycytidine treatment in hepatocellular carcinoma cells. |
| title_fullStr | Identification and characterization of microRNA candidates that are upregulated by 5-aza-2’-deoxycytidine treatment in hepatocellular carcinoma cells. |
| title_full_unstemmed | Identification and characterization of microRNA candidates that are upregulated by 5-aza-2’-deoxycytidine treatment in hepatocellular carcinoma cells. |
| title_short | Identification and characterization of microRNA candidates that are upregulated by 5-aza-2’-deoxycytidine treatment in hepatocellular carcinoma cells. |
| title_sort | identification and characterization of microrna candidates that are upregulated by 5 aza 2 deoxycytidine treatment in hepatocellular carcinoma cells |
| topic | DRNTU::Science::Biological sciences::Genetics |
| url | http://hdl.handle.net/10356/16345 |
| work_keys_str_mv | AT lengcharmaineyongen identificationandcharacterizationofmicrornacandidatesthatareupregulatedby5aza2deoxycytidinetreatmentinhepatocellularcarcinomacells |