Coordinated mRNA and protein expression of human LAMP-1 in induction of melanogenesis after UV-B exposure and co-transfection of human tyrosinase and TRP-1 cDNAs.
In order to better understand the cascade of melanogenic events in melanocytes, this report has introduced our two recent approaches for the expression of melanogenesis/or melanosome-associated genes and encoded proteins in melanocytes (melanoma cells) after repeated exposure to UV-B and after cotransfection of two human genes, i.e., tyrosinase and tyrosinase-related protein-1 (TRP-1). Repeated exposure of UV-B (2.5-5.0 mJ/cm2) caused not only upregulation of tyrosinase and TRP-1 genes but also coordinated increase in the gene and protein synthesis expression of Lamp-1 (lysosome-associated membrane protein-1). When COS-7 kidney cells and amelanotic melanoma (C32 and SK-MEL-24) and melanotic melanoma (G361 and SK-MEL-23) cells were exposed to cotransfection of human tyrosinase and TRP-1 cDNAs, there was also an increased expression of Lamp-1 mRNA and protein along with tyrosinase activation and new melanin synthesis. Importantly, single transfectants of human tyrosinase cDNA revealed marked cellular degeneration, whereas this degeneration was not seen in single transfectants of TRP-1 cDNA or cotransfectants of human tyrosinase and TRP-1 cDNAs, indicating that TRP-1 prevented, along with Lamp-1, programmed death of melanocytes after transfection of tyrosinase gene. The coordinated expression of TRP-1 and Lamp-1 was further confirmed by antisense oligodeoxynucleotide hybridization experiment against Lamp-1 gene, showing the decreased expression of TRP-1 as identified by three different types of anti-TRP-1 monoclonal antibodies.(ABSTRACT TRUNCATED AT 250 WORDS)
Jimbow
K
K
Faculty of Medicine, University of Alberta, Edmonton, Canada.
Luo
D
D
Chen
H
H
Hara
H
H
Lee
M H
MH
D000595:Amino Acid Sequence; D015703:Antigens, CD; D001483:Base Sequence; D002460:Cell Line; D015870:Gene Expression; D006801:Humans; D007668:Kidney; D051907:Lysosomal Membrane Proteins; D008543:Melanins / Q000096:biosynthesis*; D008544:Melanocytes / Q000378:metabolism; D008545:Melanoma; D008562:Membrane Glycoproteins / Q000737:chemistry / Q000235:genetics*; D008969:Molecular Sequence Data; D014442:Monophenol Monooxygenase / Q000737:chemistry / Q000235:genetics*; D010088:Oxidoreductases; D011506:Proteins / Q000737:chemistry / Q000235:genetics*; D012333:RNA, Messenger / Q000378:metabolism*; D014162:Transfection; D014407:Tumor Cells, Cultured; D014466:Ultraviolet Rays
D015703:Antigens, CD; D051907:Lysosomal Membrane Proteins; D008543:Melanins; D008562:Membrane Glycoproteins; D011506:Proteins; D012333:RNA, Messenger; D010088:Oxidoreductases; C489799:TYRP1 protein, human; C064056:tyrosinase-related protein-1; D014442:Monophenol Monooxygenase