Effect of morpholino antisense oligonucleotide against lumican mRNA in human embryonic kidney (HEK) 293 cells.
Lumican is a member of the small-leucine-rich proteoglycan (SLRP) family and is overexpressed during wound healing of the cornea, in ischemic and reperfused heart, and in several cancer tissues. Lumican is considered to regulate the collagen fibril diameter and interfibrillar spacing. However, the effect of lumican on cell growth has not been adequately examined. In the present study, we attempted to clarify whether lumican contributes to human embryonic kidney (HEK) 293 cell growth, using the morpholino antisense oligonucleotide (m-anti oligo) against lumican mRNA. M-anti oligo is a novel oligonucleotide and exhibits a higher antisense activity, higher water solubility, and greater resistance to nucleases in target cells than phosphorothioate types of oligonucleotide. After delivery of m-anti oligo against lumican mRNA, the fluorescein 5-isothiocyanate (FITC) conjugated oligonucleotides were observed in the cytoplasm and nucleus of HEK 293 cells at 24 h by confocal laser microscopy. M-anti oligo for lumican mRNA strongly inhibited the synthesis of lumican protein in the HEK 293 cells, and the HEK cell growth rate was higher than those in the control groups. These findings may indicate that lumican protein has an inhibitory effect on HEK 293 cell growth in vitro.
Ishiwata
Toshiyuki
T
Department of Pathology II, Nippon Medical School, Tokyo, Japan. ishiwata@nms.ac.jp
Fujii
Takenori
T
Ishiwata
Shunji
S
Ikegawa
Shigeo
S
Naito
Zenya
Z
D015153:Blotting, Western; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D011508:Chondroitin Sulfate Proteoglycans / Q000502:physiology*; D016650:Fluorescein-5-isothiocyanate / Q000737:chemistry; D005786:Gene Expression Regulation / Q000187:drug effects*; D006801:Humans; D007632:Keratan Sulfate / Q000502:physiology*; D007668:Kidney / Q000196:embryology; D000071234:Lumican; D018613:Microscopy, Confocal; D009025:Morpholines / Q000737:chemistry; D016376:Oligonucleotides, Antisense / Q000737:chemistry / Q000494:pharmacology*; D012333:RNA, Messenger / Q000235:genetics*
D011508:Chondroitin Sulfate Proteoglycans; C000604306:LUM protein, human; D000071234:Lumican; D009025:Morpholines; D016376:Oligonucleotides, Antisense; D012333:RNA, Messenger; D007632:Keratan Sulfate; D016650:Fluorescein-5-isothiocyanate