In vitro study examining the effect of sub-lethal QS 755 nm lasers on the expression of p16INK4a on melanoma cell lines

Abstract

Background and Objectives: Q-switched lasers had been used in the treatment of lentigo maligna but their role remains controversial. While previous studies have addressed the change in adhesion molecule expression after sub-lethal laser damage, no study has addressed the impact of sub-lethal laser damage at a molecular level. The p16 gene has been proposed as the candidate gene for melanoma. Our objective is to examine the effect of sub-lethal laser damage on p16 expression in melanoma cell lines. Study Design/Materials and Methods: Three human melanoma cell lines - HTB 66, Sk-mel-24 (HTB 71), and G361 - were irradiated by a Q-switched 755 nm Alexandrite laser at fluencies that ranged from 0.85 to 2.0 J/cm2. HTB 66 was the only cell line with significant expression of p16INK4a while the other two cells lines were p16INK4a negative and served as negative control. Protein and mRNA expression for p16 were assessed by flow cytometry and RT-PCR, respectively. Results: The level of p16INK4a protein in cell line HTB 66 increased significantly after laser irradiation as compared with non-irradiated cells. The level of p16INK4a protein did not change in p16INK4a-negative cell lines (Sk-mel-24 and G361). However, there was only a slight increase in the percentage of G0/G1 phase cells. Conclusions: Sub-lethal laser damage could increase DNA damage leading to an increase in p16 expression, and such effect would be particularly undesirable for patients with p16 mutation. Further studies are warranted to examine the role of sub-lethal laser damage in inducing p16 mutation. © 2003 Wiley-Liss, Inc.