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目的探讨阻断细胞外信号调节激酶通路后对5-氨基酮戊酸光动力疗法(ALA-PDT)杀伤人皮肤鳞状细胞癌细胞株(SCL-1)细胞的影响。方法将SCL-1细胞分为空白对照组、激光照射组、ALA组、ALA-PDT组及ERK阻断剂组。Western蛋白印迹法检测各组细胞干预30min,60min,90min后细胞外信号调节激酶1/2(ERK1/2)蛋白产物及磷酸化细胞外信号调节激酶1/2(p-ERK1/2)蛋白产物的表达;用MTT(噻唑蓝)酶联免疫法分别检测各组细胞在干预后24h,48h,72h的光密度值,计算各组细胞存活率;用Annexin V-FITC/PI双染流式细胞术检测各组细胞干预后24h,48h,72h细胞的凋亡率。结果与其余各组相比,ERK阻断剂组p-ERK1/2蛋白表达水平有明显下降;ERK阻断剂组细胞的存活率显著下降;ERK阻断剂组细胞的早期凋亡率明显增高,差异均有统计学意义(P<0.05)。结论阻断ERK通路可能成为增强ALA-PDT杀伤皮肤鳞癌细胞新的治疗靶点。
Objective To investigate the effect of 5-aminolevulinic acid photodynamic therapy (ALA-PDT) on human skin squamous cell carcinoma cell line (SCL-1) after blocking the extracellular signal-regulated kinase pathway. Methods SCL-1 cells were divided into blank control group, laser irradiation group, ALA group, ALA-PDT group and ERK blocker group. Western blotting was used to detect the expression of extracellular signal-regulated kinase 1/2 (ERK1 / 2) protein and phosphorylated extracellular signal-regulated kinase 1/2 (p-ERK1 / 2) protein after 30, 60, . The optical density of each group was detected by MTT (thiazolyl-blue) enzyme-linked immunosorbent assay at 24h, 48h, 72h after intervention. Cell viability was calculated by Annexin V-FITC / PI double staining flow cytometry The apoptosis rate of cells in each group at 24h, 48h, 72h after intervention was detected. Results Compared with the other groups, the expression of p-ERK1 / 2 protein in ERK-inhibitor group was significantly decreased; the survival rate of ERK-inhibitor group was significantly decreased; the early apoptosis rate of ERK-inhibitor group was significantly increased , The differences were statistically significant (P <0.05). Conclusion Blocking the ERK pathway may be a new therapeutic target for enhancing the cytotoxicity of ALA-PDT on skin squamous cell carcinoma.