Transcriptional Regulation of Liver-Type OATP1B3 (Lt-OATP1B3) and Cancer-Type OATP1B3 (Ct-OATP1B3) Studied in Hepatocyte-Derived and Colon Cancer-Derived Cell Lines
Due to alternative splicing, the <i>SLCO1B3</i> gene encodes two protein variants; the hepatic uptake transporter liver-type OATP1B3 (Lt-OATP1B3) and the cancer-type OATP1B3 (Ct-OATP1B3) expressed in several cancerous tissues. There is limited information about the cell type-specific tra...
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MDPI AG
2023-02-01
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| Online Access: | https://www.mdpi.com/1999-4923/15/3/738 |
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| author | Bastian Haberkorn Dennis Löwen Lukas Meier Martin F. Fromm Jörg König |
| author_facet | Bastian Haberkorn Dennis Löwen Lukas Meier Martin F. Fromm Jörg König |
| author_sort | Bastian Haberkorn |
| collection | DOAJ |
| description | Due to alternative splicing, the <i>SLCO1B3</i> gene encodes two protein variants; the hepatic uptake transporter liver-type OATP1B3 (Lt-OATP1B3) and the cancer-type OATP1B3 (Ct-OATP1B3) expressed in several cancerous tissues. There is limited information about the cell type-specific transcriptional regulation of both variants and about transcription factors regulating this differential expression. Therefore, we cloned DNA fragments from the promoter regions of the <i>Lt-SLCO1B3</i> and the <i>Ct-SLCO1B3</i> gene and investigated their luciferase activity in hepatocellular and colorectal cancer cell lines. Both promoters showed differences in their luciferase activity depending on the used cell lines. We identified the first 100 bp upstream of the transcriptional start site as the core promoter region of the <i>Ct-SLCO1B3</i> gene. In silico predicted binding sites for the transcription factors ZKSCAN3, SOX9 and HNF1α localized within these fragments were further analyzed. The mutagenesis of the ZKSCAN3 binding site reduced the luciferase activity of the <i>Ct-SLCO1B3</i> reporter gene construct in the colorectal cancer cell lines DLD1 and T84 to 29.9% and 14.3%, respectively. In contrast, using the liver-derived Hep3B cells, 71.6% residual activity could be measured. This indicates that the transcription factors ZKSCAN3 and SOX9 are important for the cell type-specific transcriptional regulation of the <i>Ct-SLCO1B3</i> gene. |
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| issn | 1999-4923 |
| language | English |
| last_indexed | 2024-03-11T06:02:36Z |
| publishDate | 2023-02-01 |
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| spelling | doaj.art-c05c60f201474b81b3d9b9b6272fb02d2023-11-17T13:13:42ZengMDPI AGPharmaceutics1999-49232023-02-0115373810.3390/pharmaceutics15030738Transcriptional Regulation of Liver-Type OATP1B3 (Lt-OATP1B3) and Cancer-Type OATP1B3 (Ct-OATP1B3) Studied in Hepatocyte-Derived and Colon Cancer-Derived Cell LinesBastian Haberkorn0Dennis Löwen1Lukas Meier2Martin F. Fromm3Jörg König4Institute of Experimental and Clinical Pharmacology and Toxicology, Friedrich-Alexander-Universität Erlangen-Nürnberg (FAU), 91054 Erlangen, GermanyInstitute of Experimental and Clinical Pharmacology and Toxicology, Friedrich-Alexander-Universität Erlangen-Nürnberg (FAU), 91054 Erlangen, GermanyInstitute of Experimental and Clinical Pharmacology and Toxicology, Friedrich-Alexander-Universität Erlangen-Nürnberg (FAU), 91054 Erlangen, GermanyInstitute of Experimental and Clinical Pharmacology and Toxicology, Friedrich-Alexander-Universität Erlangen-Nürnberg (FAU), 91054 Erlangen, GermanyInstitute of Experimental and Clinical Pharmacology and Toxicology, Friedrich-Alexander-Universität Erlangen-Nürnberg (FAU), 91054 Erlangen, GermanyDue to alternative splicing, the <i>SLCO1B3</i> gene encodes two protein variants; the hepatic uptake transporter liver-type OATP1B3 (Lt-OATP1B3) and the cancer-type OATP1B3 (Ct-OATP1B3) expressed in several cancerous tissues. There is limited information about the cell type-specific transcriptional regulation of both variants and about transcription factors regulating this differential expression. Therefore, we cloned DNA fragments from the promoter regions of the <i>Lt-SLCO1B3</i> and the <i>Ct-SLCO1B3</i> gene and investigated their luciferase activity in hepatocellular and colorectal cancer cell lines. Both promoters showed differences in their luciferase activity depending on the used cell lines. We identified the first 100 bp upstream of the transcriptional start site as the core promoter region of the <i>Ct-SLCO1B3</i> gene. In silico predicted binding sites for the transcription factors ZKSCAN3, SOX9 and HNF1α localized within these fragments were further analyzed. The mutagenesis of the ZKSCAN3 binding site reduced the luciferase activity of the <i>Ct-SLCO1B3</i> reporter gene construct in the colorectal cancer cell lines DLD1 and T84 to 29.9% and 14.3%, respectively. In contrast, using the liver-derived Hep3B cells, 71.6% residual activity could be measured. This indicates that the transcription factors ZKSCAN3 and SOX9 are important for the cell type-specific transcriptional regulation of the <i>Ct-SLCO1B3</i> gene.https://www.mdpi.com/1999-4923/15/3/738Lt-OATP1B3Ct-OATP1B3<i>Lt-SLCO1B3</i><i>Ct-SLCO1B3</i>colorectal carcinomaZKSCAN3 |
| spellingShingle | Bastian Haberkorn Dennis Löwen Lukas Meier Martin F. Fromm Jörg König Transcriptional Regulation of Liver-Type OATP1B3 (Lt-OATP1B3) and Cancer-Type OATP1B3 (Ct-OATP1B3) Studied in Hepatocyte-Derived and Colon Cancer-Derived Cell Lines Pharmaceutics Lt-OATP1B3 Ct-OATP1B3 <i>Lt-SLCO1B3</i> <i>Ct-SLCO1B3</i> colorectal carcinoma ZKSCAN3 |
| title | Transcriptional Regulation of Liver-Type OATP1B3 (Lt-OATP1B3) and Cancer-Type OATP1B3 (Ct-OATP1B3) Studied in Hepatocyte-Derived and Colon Cancer-Derived Cell Lines |
| title_full | Transcriptional Regulation of Liver-Type OATP1B3 (Lt-OATP1B3) and Cancer-Type OATP1B3 (Ct-OATP1B3) Studied in Hepatocyte-Derived and Colon Cancer-Derived Cell Lines |
| title_fullStr | Transcriptional Regulation of Liver-Type OATP1B3 (Lt-OATP1B3) and Cancer-Type OATP1B3 (Ct-OATP1B3) Studied in Hepatocyte-Derived and Colon Cancer-Derived Cell Lines |
| title_full_unstemmed | Transcriptional Regulation of Liver-Type OATP1B3 (Lt-OATP1B3) and Cancer-Type OATP1B3 (Ct-OATP1B3) Studied in Hepatocyte-Derived and Colon Cancer-Derived Cell Lines |
| title_short | Transcriptional Regulation of Liver-Type OATP1B3 (Lt-OATP1B3) and Cancer-Type OATP1B3 (Ct-OATP1B3) Studied in Hepatocyte-Derived and Colon Cancer-Derived Cell Lines |
| title_sort | transcriptional regulation of liver type oatp1b3 lt oatp1b3 and cancer type oatp1b3 ct oatp1b3 studied in hepatocyte derived and colon cancer derived cell lines |
| topic | Lt-OATP1B3 Ct-OATP1B3 <i>Lt-SLCO1B3</i> <i>Ct-SLCO1B3</i> colorectal carcinoma ZKSCAN3 |
| url | https://www.mdpi.com/1999-4923/15/3/738 |
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