摘要
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 explore the effect of blocking extracellular signal-regulated kinase（ERK1/2） pathway on the duration of 5-aminolevulinic acid（ALA） photodynamic therapy （PDT） killing SCL-1 cells. Methods SCL-1 cells were divided into control group, light group, ALA group, ALA-PDT group and inhibitor of ERK group. Western blotting technique was used to detect the expression of ERK1/2 and p-ERK1/2 after interve-ning 30min, 60min and 90min of each cell groups. MTT was used to calculated the cell survival rate after optical density value of each group were obtained at 24h,48h,72h. Flow cytometry with annexin V-FITC/PI double staining technique was employed to detect apoptosis of each cell after intervening 24h,48h and 72h. Results When compared with control group, ALA group, light group, and inhibitor of ERK group, inhibi-tor of ERK group can obviously decrease the expression of phospho-ERK1/2 （ P 〈 0.05 ） and the cell survival rate （P 〈 0.05 ） ; inhibitor of ERK group can sharply increase the rate of apoptosis （ P 〈 0.05 ） . Conclusion Blocking ERK pathway may become a new therapeutic target to enhance the treatment of ALA-PDT on squamous cell carcinoma.
出处
《中国皮肤性病学杂志》
CAS
北大核心
2013年第5期435-439,共5页
The Chinese Journal of Dermatovenereology
基金
国家自然科学基金资助项目(81060181)
宁夏自然科学基金资助项目(NZ1222)
