光动力疗法对表达人乳头瘤病毒16E7蛋白的HaCaT细胞株增殖和凋亡的影响  

Effects of photodynamic therapy on the proliferation and apoptosis of human papillomavirus type 16 E7 proteinexpressing HaCaT cells

作　　者：缪飞[1] 王秀丽 吕婷[1] 石磊 张玲琳 王宏伟[1] 

机构地区：[1]复旦大学附属华东医院,上海200040 [2]上海市皮肤病医院

出　　处：《中华皮肤科杂志》2015年第10期683-686,共4页Chinese Journal of Dermatology

基　　金：国家自然科学基金（30972665）;上海市自然科学基金（08ZR1417400）;上海市卫生系统先进适宜技术推广项目（2013SYU07）;上海市卫生局科研课题计划（20124034）;上海市级医院临床科研资源共享平台项目（SHDC12007703）

摘　　要：目的探讨氨基酮戊酸光动力疗法（ALA—PDT）对表达人乳头瘤病毒16 E7蛋白的HaCaT细胞株（简称HaCaT／HPV16 E7）增殖和凋亡的影响。方法HaCaT／HPV16 E7细胞株分为4组，即空白组、单纯光敏剂组、单纯照射组（12J／cm^2）、不同光剂量（4、8、12J／cm^2）ALA—PDT组，细胞与氨基酮戊酸（ALA）共孵育5h，采用波长630nm、功率30mW／cm^2红光照射，CCK8法检测细胞24h后存活率，流式细胞仪检测细胞3h的凋亡率，显微镜观察细胞形态。结果空白组、单纯光敏剂组、单纯照射组（12J／cm^2）、不同光剂量（4、8、12J／cm^2）AIA—PDT照射细胞24h，细胞生长存活率分别为99．15％±0．64％、98．13％±0．83％、96．85％±1．37％、68．98％±1．03％、46．03％±2．96％、23．57％±3．83％，后3组ALA—PDT组与其他3组比较，差异有统计学意义（均P〈0．05）。随着光剂量升高，细胞凋亡逐渐增加，细胞形态发生明显改变，细胞皱缩。结论ALA光动力抑制HPV感染细胞增殖并促进细胞发生凋亡，在一定光剂量范围内，呈剂量依赖性。Objective To explore the effects of aminolevulinic acid-based photodynamic therapy （ALA-PDT） on the proliferation and apoptosis of HaCaT cells stably expressing human papillomavirus type 16 E7 protein （HaCaT/ HPV16 E7 cells）. Methods Cultured HaCaT/HPV16 E7 cells were divided into several groups： blank control group receiving no treatment, ALA group treated with ALA alone, irradiation group irradiated with 630-nm red laser （30 mW/cm^2, 12 J/cm^2）, ALA-PDT groups pretreated with ALA for 5 hours followed by 630-nm red laser radiation at 4, 8, 12 J/cm^2 respectively. CCK8 assay was performed to determine the survival rate of cells at 24 hours after PDT, and flow cytometry and eonfoeal microscopy were conducted to detect cell apoptosis and observe cell morphology respectively at 3 hours. Results At 24 hours, the survival rate of cells was 68.98% ± 1.03%, 46.03% ±2.96% and 23.57% ±3.83% in the 4-, 8- and 12-J/cm^2 ALA-PDT groups respectively, significantly lower than that in the blank control group, ALA group and irradiation group （99.15% ±0.64%, 98.13% ±0.83% and 96.85% ±1.37% respectively, all P 〈 0. 05）. With the increase in radiation dose, cell apoptosis was accelerated with obvious morphological changes and shrinkage of cells in the ALA-PDT groups. Conclusion ALA-PDT can inhibit the proliferation, and promote the apoptosis of HPV-infeeted HaCaT cells in a dose-dependent manner within a certain range of radiation dose.

关 键 词：氨基酮戊酸 光化学疗法 细胞增殖 细胞凋亡 α乳头状瘤病毒属 人乳头瘤病毒16 E7蛋白 

分 类 号：R751[医药卫生—皮肤病学与性病学]