Surfactant protein A-directed toxin gene kills lung cancer cells in vitro.
Human surfactant protein A (SPA) expression is considered a marker of respiratory epithelial differentiation. Non-small cell lung cancers (NSCLC) are respiratory epithelial derivatives, and it was previously shown that a minority of these cancers expressed SPA, presumably a consequence of their respiratory epithelial origin. In the studies reported here, SPA-I gene transcriptional regulatory sequences were localized to a 2.75-kb genomic 5'-flanking region fragment obtained by screening a human genomic library. The 2.75-kb fragment was used to direct a luciferase coding sequence transcriptionally within a plasmid construct. In plasmid transduction experiments, the SPA-directed luciferase plasmid produced significant luciferase activity in the SPA-expressing NSCLC cell line, H441, but only background levels in the non-SPA-expressing A549 cells. Because Northern blot analysis of resected NSCLC showed that the majority expressed SPA, an SPA-transcriptional targeting strategy was investigated using chimeric toxin genes comprising the coding sequence for herpes simplex virus thymidine kinase (HSV-TK) under transcriptional control of SPA or SV40 regulatory sequences. As expected, transduction of the constitutive, SV40-directed plasmid followed by ganciclovir treatment reduced numbers of both the A549 and H441 cells. In contrast, the SPA-directed plasmid reduced only the SPA-expressing H441 cells and had no significant effect on the A549 cells. The results of these in vivo experiments suggest the concept of transcriptionally directing toxin genes with SPA can produce targeted toxicity in NSCLC.
Smith
M J
MJ
Division of Pulmonary and Critical Care Medicine, UAB School of Medicine, Birmingham, AL 35294.
Rousculp
M D
MD
Goldsmith
K T
KT
Curiel
D T
DT
Garver
R I
RI
D002289:Carcinoma, Non-Small-Cell Lung / Q000628:therapy*; D015316:Genetic Therapy; D006801:Humans; D008175:Lung Neoplasms / Q000601:surgery / Q000628:therapy*; D010957:Plasmids; D011510:Proteolipids / Q000235:genetics*; D037661:Pulmonary Surfactant-Associated Proteins; D011663:Pulmonary Surfactants / Q000235:genetics*; D012045:Regulatory Sequences, Nucleic Acid; D018139:Simplexvirus / Q000201:enzymology; D013937:Thymidine Kinase / Q000235:genetics / Q000627:therapeutic use; D014118:Toxins, Biological / Q000235:genetics* / Q000627:therapeutic use; D014158:Transcription, Genetic; D014407:Tumor Cells, Cultured
D011510:Proteolipids; D037661:Pulmonary Surfactant-Associated Proteins; D011663:Pulmonary Surfactants; D014118:Toxins, Biological; D013937:Thymidine Kinase