Differential in vitro sensitivity of oral precancerous and squamous cell carcinoma cell lines to 5-aminolevulinic acid-mediated photodynamic therapy.

Abstract

OBJECTIVES\nThe clinical effect of 5-aminolevulinic acid-mediated photodynamic therapy (ALA-PDT) may be correlated with the degree of dysplasia of cancer tissues, but much is still unknown regarding the differences in its effectiveness, especially in oral cancer and precancerous lesions. The aim of this study is to compare the effects of ALA-PDT on a human oral precancerous cell line (DOK) and an oral squamous cell carcinoma cell line (CAL-27).\n\n\nMETHODS\nFirst, we explored the dose- and time-dependent responses of DOK and CAL-27 cells to ALA-PDT. DOK and CAL-27 cells were incubated with various concentrations of ALA (from 0.25 to 2\u2009mM), followed by PDT using laser irradiation at 635\u2009nm. The resulting photocytotoxicity was assessed in both cell lines using MTT assays. Further, apoptosis was assessed using flow cytometry, reactive oxygen species (ROS) generation was evaluated with 2,7-dichlorofluorescein diacetate (DCFH2-DA), and the response to treatment was examined via RT-qPCR and Western blotting to measure the mRNA and protein expression levels of matrix metallopeptidase 2 (MMP-2) and MMP-9.\n\n\nRESULTS\nALA-PDT inhibited the proliferation of DOK and CAL-27 cells in a dose- and time-dependent manner. Dose-effect and inhibition-time relationships were also found. The rates of DOK and CAL-27 cell apoptosis when the ALA dose was 1\u2009mM were 30.66\u2009±\u20093.10 % and 75.40\u2009±\u20091.29 %, respectively (P\u2009<\u20090.01). Following PDT, compared with DOK cells, the ROS level in CAL-27 cells was significantly increased and was correlated with an increase in the ALA concentration. Mechanistically, both the mRNA and protein expression levels of MMP-2 and MMP-9 were found to be regulated in both cell types after ALA-PDT.\n\n\nCONCLUSION\nALA-PDT effectively killed DOK and CAL-27 cells in a dose- and time-dependent manner in vitro. However, under the same conditions, the susceptibilities of these cell lines to ALA-PDT were different. Further studies are necessary to confirm whether this difference is present in clinical oral cancer and precancerous lesions.