Transcriptional targeting of gene expression in breast cancer by the promoters of protein regulator of cytokinesis 1 and ribonuclease reductase 2

Abstract

For cancer gene therapy, cancer-specific overexpression of a therapeutic gene is required to reduce side effects derived from expression of the gene in normal cells. To develop such an expression vector, we searched for genes over-expressed and/or specifically expressed in cancer cells using bioinformatics and have selected genes coding for protein regulator of cytokinesis 1 (PRC1) and ribonuclease reductase 2 (RRM2) as candidates. Their cancer-specific expressions were confirmed in both breast cancer cell lines and patient tissues. We compared each promoter's cancer-specific activity in the breast normal and cancer cell lines using the luciferase gene as a reporter and confirmed cancer-specific expression of both PRC1 and RRM2 promoters. To test activities of these promoters in viral vectors, the promoters were also cloned into an adeno-associated viral (AAV) vector containing green fluorescence protein (GFP) as the reporter. The GFP expression levels by these promoters were various depending on cell lines tested and, in MDA-MB-231 cells, GFP activities derived from the PRC1 and RRM2 promoters were as strong as that from the cytomegalovirus (CMV) promoter. Our result showed that a vector containing the PRC1 or RRM2 promoter could be used for breast cancer specific overexpression in gene therapy.

Expression of ribonucleotide reductase M2 subunit in gastric cancer and effects of RRM2 inhibition in vitro

“…1). There have been few studies that have examined RRM2 protein expression in human cancer tissues [18][19][20]. Kolesar expression in only four cases of "esophageal and gastric cancers" and found that those cancers overexpressed RRM2 protein when compared with prostate cancer [19].…”

Transcription of the protein kinase C-δ gene is activated by JNK through c-Jun and ATF2 in response to the anticancer agent doxorubicin

“…NIH 3T3 cells were seeded into a 12-well plate and transfected with 0.5 μg of PKCδ promoter-reporter construct using Lipofectamine 2000 as described previously (Yun et al, 2008). To monitor the transfection efficiency, the pRL-null plasmid (5 ng) encoding Renilla luciferase was included in all samples.…”

Identification and functional characterization of glioma-specific promoters and their application in suicide gene therapy

“…In addition, the efficacy of hTERT promoter gene therapy limits further application, because its activity is not high [13]. Therefore, identification of novel promoters with tumor-specific and strong activity remains a goal for the development of tumor-specific gene therapy.…”