Reference - Detail
| RRC ID | 44839 |
|---|---|
| Author | Danda R, Krishnan G, Ganapathy K, Krishnan UM, Vikas K, Elchuri S, Chatterjee N, Krishnakumar S. |
| Title | Targeted expression of suicide gene by tissue-specific promoter and microRNA regulation for cancer gene therapy. |
| Journal | PLoS One |
| Abstract |
In order to realise the full potential of cancer suicide gene therapy that allows the precise expression of suicide gene in cancer cells, we used a tissue specific Epithelial cell adhesion molecule (EpCAM) promoter (EGP-2) that directs transgene Herpes simplex virus-thymidine kinase (HSV-TK) expression preferentially in EpCAM over expressing cancer cells. EpCAM levels are considerably higher in retinoblastoma (RB), a childhood eye cancer with limited expression in normal cells. Use of miRNA regulation, adjacent to the use of the tissue-specific promoter, would provide the second layer of control to the transgene expression only in the tumor cells while sparing the normal cells. To test this hypothesis we cloned let-7b miRNA targets in the 3'UTR region of HSV-TK suicide gene driven by EpCAM promoter because let-7 family miRNAs, including let-7b, were found to be down regulated in the RB tumors and cell lines. We used EpCAM over expressing and let-7 down regulated RB cell lines Y79, WERI-Rb1 (EpCAM (+ve)/let-7b(down-regulated)), EpCAM down regulated, let-7 over expressing normal retinal Müller glial cell line MIO-M1(EpCAM (-ve)/let-7b(up-regulated)), and EpCAM up regulated, let-7b up-regulated normal thyroid cell line N-Thy-Ori-3.1(EpCAM (+ve)/let-7b(up-regulated)) in the study. The cell proliferation was measured by MTT assay, apoptosis was measured by probing cleaved Caspase3, EpCAM and TK expression were quantified by Western blot. Our results showed that the EGP2-promoter HSV-TK (EGP2-TK) construct with 2 or 4 copies of let-7b miRNA targets expressed TK gene only in Y79, WERI-Rb-1, while the TK gene did not express in MIO-M1. In summary, we have developed a tissue-specific, miRNA-regulated dual control vector, which selectively expresses the suicide gene in EpCAM over expressing cells. |
| Volume | 8(12) |
| Pages | e83398 |
| Published | 2013-1-1 |
| DOI | 10.1371/journal.pone.0083398 |
| PII | PONE-D-13-27551 |
| PMID | 24391761 |
| PMC | PMC3877029 |
| MeSH | Antigens, Neoplasm / genetics Breast Neoplasms / genetics Breast Neoplasms / pathology Breast Neoplasms / therapy Cell Adhesion Molecules / genetics Cell Death / drug effects Cell Death / genetics Cell Line Cell Line, Tumor Ependymoglial Cells / metabolism Epithelial Cell Adhesion Molecule Eye Neoplasms / genetics* Eye Neoplasms / pathology Eye Neoplasms / therapy* Female Ganciclovir / pharmacology Gene Expression Gene Targeting / methods Genes, Transgenic, Suicide* Genetic Therapy / methods* Humans MCF-7 Cells MicroRNAs / genetics* Middle Aged Promoter Regions, Genetic RNA, Neoplasm / genetics Retinoblastoma / genetics* Retinoblastoma / pathology Retinoblastoma / therapy* Simplexvirus / enzymology Simplexvirus / genetics Thymidine Kinase / genetics Tissue Distribution |
| IF | 2.74 |
| Times Cited | 16 |
| WOS Category | MEDICINE, RESEARCH & EXPERIMENTAL |
| Altmetric score |
オルトメトリクス指標項目
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| The most frequently cited source | Patent(IFI CLAIMS) |
| Total number of mentions | 2 |
| Altmetric score changes over past 6months | 3.0 |
| Resource | |
| Human and Animal Cells | Y79(RCB1645) WERI-Rb-1(RCB2146) MDA-MB-453(RCB1192) MCF7(RCB1904) |