Decreased proliferation of human melanoma cell lines caused by antisense RNA against translation factor eIF-4A1.
Control of translation initiation was recognised as a critical checkpoint for cell proliferation and tumorigenesis. In human melanoma cells, we have previously reported consistent overexpression of translation initiation factor eIF-4A1. Here, we investigated by transfection of antisense constructs its significance for the control of melanoma cell growth. The tetracycline-inducible expression system was established in melanoma cells, and three fragments of the 5'-, central-, and 3'-portion of the eIF-4A1 cDNA were subcloned in antisense and in sense orientation after a tetracycline inducible promoter. Significant proliferation decrease was obtained after transient transfection and induction of antisense RNA directed against the 5'- and the central portion (up to 10%), whereas, no effects were seen after induction of the 3'-fragment and the sense controls. Cell clones stably transfected with the central antisense fragment revealed after doxycycline induction reduced expression of endogeneous eIF-4A1 mRNA correlated with decreased proliferation rates (up to 6%). These data demonstrate the applicability of antisense strategies against translation factors in melanoma cells. Translation initiation factor eIF-4A1 contributes to the control of melanoma cell proliferation and may be taken into consideration when scheduling new therapeutic approaches targeting the translational control.
Eberle
J
J
Department of Dermatology, University Medical Center Benjamin Franklin, The Free University of Berlin, Fabeckstrasse 60-62, 14195 Berlin, Germany. eberle@ukbf.fu-berlin.de
Fecker
L F
LF
Bittner
J-U
JU
Orfanos
C E
CE
Geilen
C C
CC
D000900:Anti-Bacterial Agents / Q000494:pharmacology; D002455:Cell Division; D015536:Down-Regulation; D039601:Eukaryotic Initiation Factor-4A; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D008545:Melanoma / Q000378:metabolism / Q000473:pathology*; D010448:Peptide Initiation Factors / Q000235:genetics*; D011132:Polyribosomes / Q000378:metabolism; D016372:RNA, Antisense / Q000502:physiology*; D012333:RNA, Messenger / Q000378:metabolism; D012878:Skin Neoplasms / Q000378:metabolism / Q000473:pathology*; D013754:Tetracyclines; D014162:Transfection; D014407:Tumor Cells, Cultured
D000900:Anti-Bacterial Agents; D010448:Peptide Initiation Factors; D016372:RNA, Antisense; D012333:RNA, Messenger; D013754:Tetracyclines; D039601:Eukaryotic Initiation Factor-4A