Sub-lethal effects of exposing the human melanoma cell line SKmel-23 to 532 nm laser light.
The human melanoma cell line SKmel-23 has been used to investigate the sub-lethal damage that can occur as a result of exposing melanin containing cells to light (532 nm) from a frequency doubled Q-switched (Nd:YAG) laser. A dose response curve was obtained, which indicates that at energy levels of 0.6 J/cm2 and below no effect on either the viability or growth rate of the cell line was observed. Above this, cells rapidly died and at an energy level of 2.0 J/cm2, only approximately 15% of cells survived. This contrasts with the effects on the G361 melanoma line, which contains far less melanosomes, as an LD50 for this cell line was approximately 5.5 J/cm2. Exposing SKmel-23 cells to 0.4 J/cm2 of 532 nm light results in a diminution of the number of melanosomes within cells as well as a marked decrease in melanin content, as determined by spectrophotometric assay and electron microscopy. Using the reverse transcriptase polymerase chain reaction technique, the reduction in melanin content of the cells was accompanied by a selective decrease in mRNA coding for tyrosinase, the first enzyme in the biosynthetic pathway for melanin. No decrease in the mRNA coding for the GAPDH protein was observed. Our finding has implications for understanding the control processes that regulate the melanin content of cells and suggests that the model described can be used to further investigate changes that may occur in cells as a result of their exposure to sub-lethal levels of laser light.
Zhu
N W
NW
Department of Plastic Surgery, Frenchay Hospital, Bristol, UK. N.Zhu@bristol.ac.uk
Kenealy
J
J
Burd
A
A
Gradidge
T
T
Warr
R
R
Rigby
H S
HS
Kemshead
J T
JT
D002455:Cell Division / Q000528:radiation effects; D002460:Cell Line; D002470:Cell Survival / Q000528:radiation effects*; D004307:Dose-Response Relationship, Radiation; D006801:Humans; D007700:Kinetics; D007834:Lasers; D008027:Light; D008543:Melanins / Q000096:biosynthesis; D008545:Melanoma / Q000648:ultrastructure; D014442:Monophenol Monooxygenase / Q000096:biosynthesis; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000096:biosynthesis; D012878:Skin Neoplasms / Q000648:ultrastructure; D014158:Transcription, Genetic / Q000528:radiation effects*; D014407:Tumor Cells, Cultured
D008543:Melanins; D012333:RNA, Messenger; D014442:Monophenol Monooxygenase