Fas ligand downregulation with antisense oligonucleotides in cells and in cultured tissues of normal skin epidermis and basal cell carcinoma.
Fas ligand (FasL), a member of the tumor necrosis factor family, induces apoptosis upon interaction with Fas-receptor-expressing cells. FasL normally plays an important immune regulatory role, but it can also cause severe skin diseases if overexpressed and it may serve some tumors for immune evasion. Thus, in situ inhibition of FasL expression with antisense oligonucleotides in patients may be a novel approach to overcome its pathogenic role. We designed and evaluated 15 phosphorothioate antisense oligonucleotides directed against different regions of the human FasL mRNA. They exhibited different inhibitory activities on FasL expression in HEK293 cells. The most potent antisense oligonucleotide, ASO8, specifically downregulated 90% FasL expression at the protein level and 80% at the mRNA level. FasL downregulation reduced the effector function of HEK293 cells toward Fas receptor positive target cells. Further studies demonstrated that ASO8 efficiently inhibited FasL synthesis in split skin and basal cell carcinoma tissue. Our results show that the modulation of FasL expression by antisense oligonucleotides is possible in cells as well as tissue and indicate that antisense oligonucleotides may provide a promising strategy for the therapy of FasL-mediated disorders.
Ji
Jingmin
J
Institute for Medical Microbiology, University of Basel, and Department of Dermatology, University of Basel, Switzerland.
Wernli
Marion
M
Buechner
Stanislaw
S
Erb
Peter
P
D017209:Apoptosis / Q000187:drug effects / Q000502:physiology; D002280:Basal Cell Carcinoma / Q000378:metabolism*; D002460:Cell Line; D046508:Culture Techniques; D015536:Down-Regulation; D004817:Epidermis / Q000378:metabolism*; D053222:Fas Ligand Protein; D006801:Humans; D008562:Membrane Glycoproteins / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism* / Q000502:physiology; D016376:Oligonucleotides, Antisense / Q000494:pharmacology*; D012867:Skin / Q000378:metabolism; D012878:Skin Neoplasms / Q000378:metabolism*
C505661:FASLG protein, human; D053222:Fas Ligand Protein; D008562:Membrane Glycoproteins; D016376:Oligonucleotides, Antisense