中华皮肤科杂志 ›› 2015, Vol. 48 ›› Issue (10): 683-686.

• 论著 • 上一篇    下一篇

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

缪飞1,王秀丽2,吕婷3,石磊2,张玲琳2,王宏伟4   

  1. 1. 复旦大学附属华东医院
    2. 上海市皮肤病医院
    3. 上海市复旦大学附属华东医院
    4. 复旦大学附属华东医院皮肤科
  • 收稿日期:2015-01-06 修回日期:2015-06-10 发布日期:2015-09-28
  • 通讯作者: 王宏伟 E-mail:hongweiwang2005@aliyun.com
  • 基金资助:

    ALA光动力诱导HPV16E7转染角质形成细胞凋亡机制研究;ALA光动力对HPV16E7转染角质形成细胞作用机制研究;ALA光动力对早期皮肤鳞癌免疫学干预效应研究;上海市级医院临床科研资源共享平台项目

Effects of photodynamic therapy on the proliferation and apoptosis of human papillomavirus type 16 E7 protein-expressing HaCaT cellsa

Fei MIAO 2, 2,lei SHI2, 2,wang hong-wei   

  • Received:2015-01-06 Revised:2015-06-10 Published:2015-09-28
  • Contact: wang hong-wei E-mail:hongweiwang2005@aliyun.com

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(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5 h,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3 h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。

Abstract:

Miao Fei *, Wang Xiuli, Lyu Ting, Shi Lei, Zhang Linglin, Wang Hongwei. *Huadong Hospital Affiliated to Fudan University, Shanghai 200040, China Corresponding author: Wang Hongwei, Email: hongweiwang2005@aliyun.com 【Abstract】 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 cytometry 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.

引用本文

缪飞 王秀丽 吕婷 石磊 张玲琳 王宏伟. 光动力疗法对表达人乳头瘤病毒16 E7蛋白的HaCaT细胞株增殖和凋亡的影响[J]. 中华皮肤科杂志, 2015,48(10):683-686. doi:

Fei MIAO lei SHI wang hong-wei. Effects of photodynamic therapy on the proliferation and apoptosis of human papillomavirus type 16 E7 protein-expressing HaCaT cellsa[J]. Chinese Journal of Dermatology, 2015, 48(10): 683-686.doi: