氨基酮戊酸光动力疗法治疗SKH-1小鼠皮肤鳞状细胞癌作用机制研究  

Mechanisms underlying the effect of 5-aminolevulinic acid photodynamic therapy on cutaneous squamous cell carcinoma in SKH-1 mice

作　　者：李晶晶 吕婷 王宏伟 王佩茹 王秀丽 

机构地区：[1]上海市皮肤病医院,200050

出　　处：《中华皮肤科杂志》2014年第3期181-185,共5页Chinese Journal of Dermatology

基　　金：国家自然科学基金（81272990）;上海市自然科学基金（11ZR1432800）

摘　　要：目的探讨氨基酮戊酸光动力疗法（ALA—PDT）对小鼠皮肤鳞状细胞癌（SCC）的作用机制。方法先建立SKH-1无毛小鼠SCC模型，ALA—PDT治疗前即对照组3只，ALA—PDT治疗后1、3、6、12、24h组各3只。以透射电镜及TUNEL法观察ALA—PDT治疗后1～24hSCC组织肿瘤细胞死亡方式（坏死、凋亡、自噬）；以免疫组化技术检测治疗后7dSCC组织细胞自噬标记物LC3B、肿瘤局部微血管CD34、肿瘤局部免疫细胞浸润包括树突细胞CDla、CD4’T淋巴细胞和CD8’T淋巴细胞表达的变化。采用SPSS17．0软件进行统计学分析，两样本均数比较采用t检验。结果透射电镜观察发现，ALA—PDT治疗后24h，肿瘤细胞逐渐发生坏死和凋亡，且以细胞坏死为主，巨噬细胞中可见自噬小体。TUNEL检测显示，肿瘤细胞凋亡显著增加（ALA—PDT治疗前及治疗后24h分别为2．00±0．69和7．30±2．18，P〈0．05）。免疫组化显示，与治疗前比较，ALA—PDT治疗后7d，CD34表达显著下降（分别为19．00±2．66和1．33±0．58，P〈0．01），肿瘤局部CDla表达明显增高（分别为10．33±1．88和23．01±2．04，P〈0．05），CD4＋T细胞数（分别为12．40±2．27和28．67士1．76，P〈0．05）和CD8＋T细胞数（分别为11．67±1．45和25．79±2．37，P〈0．05）明显增多，同时肿瘤间质中LC3B表达明显增多（分别为21．44±4．3和30．6±3．21，P〈0．05）。结论ALA—PDT可通过细胞坏死和凋亡两种方式直接杀伤SCC细胞，未发现肿瘤细胞自噬性死亡；ALA—PDT可损伤SCC组织微血管内皮细胞，诱导局部树突细胞以及CD4＋、CD8＋T细胞聚集。Objective To investigate the mechanisms underlying the effect of 5-aminolevulinic acid photodynamic therapy （ALA-PDT） on cutaneous squamous cell carcinoma （SCC） in mice. Methods A model of cutaneous SCC was established in 21 SKH-1 hairless mice, which were treated with topical ALA 8% cream followed by single irradiation with He-Ne laser at a total dose of 30 J/cm2 （ALA-PDT）. Three mice were sacrificed before and at 1, 3, 6, 12, 24 hours and 7 days after the irradiation, separately, and SCC tissue was taken from the mice. Transmission electron microscopy （TEM） and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling （TUNEL） were performed to determine the pattern of tumor cell death（necrosis, apoptosis and autophagy） during 1 - 24 hours after ALA-PDT, and immunohistochemical techniques were used to estimate the expressions of LC3B and CD34 on SCC cells, as well as the quantity of CDla＋ cells, CD4＋ T and CD8＋ T lymphocytes in SCC tissue 7 days after the irradiation. Statistical analysis was done by two-sample t test using SPSS 17.0 software. Results TEM showed gradual necrosis and apoptosis （especially necrosis） of tumor cells and formation of autophagosomes in macrophages within 24 hours after ALA-PDT. The number of apoptotic ceils per high power field （＋ 400） in SCC tissue significantly increased at 24 hours compared with that before ALA-PDT （7.30 ± 2.18 vs. 2.00 ± 0.69, P 〈 0.05）. As immunohistochemistry revealed, there was a significant decrease in the number of CD34＋ cells （1.33 ± 0.58 vs. 19.00 ± 2.66, P 〈 0.01）, but a marked increase in that of CDla＋ cells （23.01 ± 2.04 vs. 10.33 ± 1.88, P 〈 0.05）, CD4＋ T cells （28.67± 1.76 vs. 12.40 ± 2.27, P 〈 0.05）, CD8＋ T cells （25.79± 2.37 vs. 11.67 ± 1.45, P 〈 0.05） and LC3B＋ interstitial cells （30.6 ± 3.21 vs. 21.44 ± 4.3, P 〈 0.05） per high power field （x 400） in SCC tissue on day 7 compared with that before ALA-PDT. Conclusions ALA-PDT may directly

关 键 词：癌 鳞状细胞 光疗法 细胞死亡 微血管损伤 免疫效应 

分 类 号：R739.5[医药卫生—肿瘤]