CAJ | 학술논문

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/HPV 16 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(12 J/cm2)、不同光剂量(4、8、12 J/cm2)ALA-PDT组,细胞与氨基酮戊酸(ALA)共孵育5h,采用波长630 nm、功率30 mW/cm2红光照射,CCK8法检测细胞24 h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(12 J/cm2)、不同光剂量(4、8、12 J/cm2)ALA-PDT照射细胞24 h,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.
목적 탐토안기동무산광동력요법(ALA-PDT)대표체인유두류병독16 E7단백적HaCaT세포주(간칭HaCaT/HPV 16 E7)증식화조망적영향.방법 HaCaT/HPV16 E7세포주분위4조,즉공백조、단순광민제조、단순조사조(12 J/cm2)、불동광제량(4、8、12 J/cm2)ALA-PDT조,세포여안기동무산(ALA)공부육5h,채용파장630 nm、공솔30 mW/cm2홍광조사,CCK8법검측세포24 h후존활솔,류식세포의검측세포3h적조망솔,현미경관찰세포형태.결과 공백조、단순광민제조、단순조사조(12 J/cm2)、불동광제량(4、8、12 J/cm2)ALA-PDT조사세포24 h,세포생장존활솔분별위99.15％±0.64％、98.13％±0.83％、96.85％±1.37％、68.98％±1.03％、46.03％±2.96％、23.57％±3.83％,후3조ALA-PDT조여기타3조비교,차이유통계학의의(균P＜0.05).수착광제량승고,세포조망축점증가,세포형태발생명현개변,세포추축.결론 ALA광동력억제HPV감염세포증식병촉진세포발생조망,재일정광제량범위내,정제량의뢰성.
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/HPVl6 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/cm2,12 J/cm2), ALA-PDT groups pretreated with ALA for 5 hours followed by 630-nm red laser radiation at 4, 8, 12 J/cm2 respectively.CCK8 assay was performed to determine the survival rate of cells at 24 hours after PDT, and flow cytometetry and confocal 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/cm2 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-infected HaCaT cells in a dose-dependent manner within a certain range of radiation dose.