The insulinoma-associated 1: a novel promoter for targeted cancer gene therapy for small-cell lung cancer.
The insulinoma-associated 1 (INSM1) gene is expressed exclusively during early embryonal development, but has been found re-expressed at high levels in neuroendocrine tumors. The regulatory region of the INSM1 gene is therefore a potential candidate for regulating expression of a therapeutic gene in transcriptionally targeted cancer gene therapy against neuroendocrine tumors. We analyzed expression of a reporter gene from a 1.7 kb region of the INSM1 promoter in a large number of small-cell lung cancer (SCLC) cell lines. This INSM1 promoter region showed very high levels of expression in most of the SCLC cell lines and expression was absent in cell lines of non-neuroendocrine origin. Inclusion of the general transcriptional enhancer from SV40 compromised the specificity of the promoter and did not enhance transcription in most of the SCLC cell lines. For comparison, the region of the gastrin releasing peptide (GRP) previously suggested for SCLC gene therapy was analyzed in a similar manner. High expression was observed for a number of cell lines, but unlike for the INSM1 promoter, reporter gene expression from the GRP promoter did not correlate to the relative GRP mRNA levels, demonstrating that this region may not contain all necessary regulatory elements. Expression of the suicide gene herpes simplex virus thymidine kinase (HSV-TK) from the INSM1 promoter in combination with treatment with the prodrug ganciclovir (GCV) caused a significant increase in GCV sensitivity specifically in INSM1-expressing cell lines. The INSM1 promoter is therefore a potential novel tool for transcriptionally targeted gene therapy for neuroendocrine tumors.
Pedersen
N
N
Department of Radiation Biology, Finsen Center, National University Hospital, Copenhagen Ø, Denmark.
Pedersen
M W
MW
Lan
M S
MS
Breslin
M B
MB
Poulsen
H S
HS
D000998:Antiviral Agents / Q000627:therapeutic use; D018288:Carcinoma, Small Cell / Q000235:genetics / Q000473:pathology / Q000628:therapy*; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D015774:Ganciclovir / Q000627:therapeutic use; D019886:Gastrin-Releasing Peptide / Q000235:genetics / Q000378:metabolism; D015972:Gene Expression Regulation, Neoplastic; D017930:Genes, Reporter; D015316:Genetic Therapy; D006801:Humans; D008156:Luciferases; D008175:Lung Neoplasms / Q000235:genetics / Q000473:pathology / Q000628:therapy*; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000378:metabolism; D012097:Repressor Proteins / Q000235:genetics* / Q000378:metabolism; D013539:Simian virus 40 / Q000235:genetics; D018139:Simplexvirus / Q000378:metabolism; D013937:Thymidine Kinase / Q000096:biosynthesis / Q000235:genetics; D014162:Transfection
D000998:Antiviral Agents; D004268:DNA-Binding Proteins; D012333:RNA, Messenger; D012097:Repressor Proteins; C087342:INSM1 protein, human; D019886:Gastrin-Releasing Peptide; D008156:Luciferases; D013937:Thymidine Kinase; D015774:Ganciclovir