紫外线诱导SKH-1无毛小鼠皮肤鳞癌模型的构建及DNA-PKcs在小鼠及成人皮肤组织的表达

姜慧; 张娟; 徐丹; 何黎

紫外线诱导SKH-1无毛小鼠皮肤鳞癌模型的构建及DNA-PKcs在小鼠及成人皮肤组织的表达

昆明医科大学第一附属医院皮肤科云南省皮肤病医院

基金项目:

基金：国家自然科学基金资助项目 (81472540, 81660518); 昆明医科大学硕士研究生创新基金资助项目 (2018S092);

计量
- 文章访问数: 2337
- HTML全文浏览量: 821
- PDF下载量: 137
- 被引次数: 0
出版历程
- 收稿日期: 2018-11-06

Construction of UV-induced Skin Squamous Cell Carcinoma Model of SKH-1 Hairless Mice and Expression of DNA-PKcs in Mice and Adult Squamous Cell Carcinoma

Funds:

基金：国家自然科学基金资助项目 (81472540, 81660518); 昆明医科大学硕士研究生创新基金资助项目 (2018S092);

摘要

摘要: 目的紫外线照射SKH-1小鼠构建皮肤鳞状细胞癌动物模型, 探讨紫外线诱发皮肤SCC发生及AK向SCC转化过程中DNA-PKcs-m TORC2/Akt信号转导通路的DNA-PKcs活化水平。方法将43只SKH-l无毛小鼠随机分成实验组 (33只) 和对照组 (10只) 。实验组采用SUV-1000日光模拟器模拟日光照射SKH-1无毛鼠, 对照组不做任何处理。在第12、18、24、28周分别处死小鼠行皮肤组织病理检查。28周取同一只小鼠背部不同皮损行组织病理检查和免疫组化检测。收集成人日光性角化病 (AK) 、皮肤鳞状细胞癌 (SCC) 、正常避光部位 (NNS) 、正常曝光部位 (NES) 皮肤组织各5例, 进行后续Western Blot验证。结果 12周后实验组部分小鼠背部皮肤增厚, 皮棘隆起;17周开始实验组小鼠背部皮肤陆续出现直径≥l mm的丘疹;照射28周后成瘤率达100%;对照组未见肿瘤形成。DNA-PKcs和p-DNA-PKcs (T2609) 均细胞定位于细胞质和细胞核, 小鼠表皮组织中DNA-PKcs表达含量增加, 统计学分析有差异 (P<0.05) , 而p-DNA-PKcs (T2609) 无差异 (P>0.05) 。人表皮组织中DNA-PKcs和p-DNA-PKcs (T2609) 表达含量均增加且有统计学分析差异 (P<0.05) 。结论成功制备小鼠AK模型和SCC模型;明确了紫外线诱导皮肤鳞状细胞癌的过程中, 可诱发DNA损伤及修复等过程。
- 紫外线 /
- 皮肤鳞状细胞癌 /
- 日光性角化病 /
- 动物 /
- 疾病模型 /
- DNA-PKcs
Abstract: Objective To construct an animal model of squamous cell carcinoma of the skin in SKH-1 mice with ultraviolet irradiation, and to investigate the DNA-PKcs activation level of DNA-PKcs-m TORC2/Akt signal transduction pathway during UV-induced skin SCC and AK to SCC transformation. Methods 43 SKH-l hairless mice were randomly divided into experimental group (33) and control group (10) .The experimental group used the SUV-1000 solar simulator to simulate sun-irradiated SKH-1 hairless mice, and the control group did not intervene. Experimental group 1 maintained irradiation for 24 weeks, and experimental group 2 maintained irradiation for 16 weeks. The mice were sacrificed at 12, 18, 24, and 28 weeks for skin histopathological examination. At 28 weeks, the same skin of the same mouse was taken for histopathological examination and immunohistochemistry.Five cases of adult solar keratosis (AK) , cutaneous squamous cell carcinoma (SCC) , normal light-protected site (NNS) , and normal exposed site (NES) were collected for Western Blot. Results After 12 weeks, some mice in the experimental group were thickened in the back skin. At 17 weeks, the papules with diameter ≥ 1 mm appeared in the back skin of experimental group. After 28 weeks of irradiation, the tumor formation rate reached100%. No tumor formation was observed in the control group. Both DNA-PKcs and p-DNA-PKcs (T2609) were localized in the cytoplasm and nucleus. Expression of DNA-PKcs increased in epidermal tissue. There was a difference in the statistical analysis of DNA-PKcs expression (P<0.05) , but there was no difference in the expression of p-DNA-PKcs (T2609) (P>0.05) .The expression levels of DNA-PKcs and p-DNA-PKcs (T2609) in human epidermal tissue increased and there were statistical differences (P<0.05) .Conclusions We successfulyl prepared of mouse AK model and SCC model. It is clear that UV-induced squamous cell carcinoma of the skin can induce DNA damage and repair.
- Ultraviolet radiation /
- Squamous cell carcinoma /
- Actinic keratosis /
- Disease models /
- Animal /
- DNA-PKcs

HTML全文

参考文献(14)

[1]	[1]ROGERS H W, WEINSTOCK M A, HARRIS A R, et al.Incidence estimate of nonmelanoma skin cancer in the United States, 2006[J].Archives of Dermatology, 2010, 146 (3) :283-287.
[2]	[2]DONALDSON M R, COLDIRON B M. No end in sight:the skin cancer epidemic continues[J].Seminars in Cutaneous Medicine and Surgery, 2011, 30 (1) :3-5.
[3]	[3]DIKA E, FANTI P A, MISCIALI C, et al. Risk of skin cancer development in 672 patients affected by actinic keratosis.Giornaleitaliano di dermatologia e venereologia:organoufficiale[J].Societaitaliana di Dermatologia E Sifilografia, 2016, 151 (6) :628-633.
[4]	[4]RASTOGI R P, RICHA, KUMAR A, et al. Molecular mechanisms of ultraviolet radiation-induced DNA damage and repair[J].Journal of Nucleic Acids, 2010, 16 (2010) :1-32.
[5]	[5]WANG Y N, FANG H, WANG H M, et al. Effect of chronic exposure to ultraviolet on skin barrier function[J].Medical Sciences, 2010, 39 (5) :517-522.
[6]	[6]SALASCHE S J. Epidemiology of actinic keratoses and squamous cell carcinoma[J].Journal of the American Academy of Dermatology, 2000, 42 ( (1 Pt 2) :4-7.
[7]	[7]KATIYAR S K. Dietary proanthocyanidins inhibit UV radiation-induced skin tumor development through functional activation of the immune system[J].Molecular Nutrition&Food Research, 2016, 60 (6) :1374-1382.
[8]	[8]D'ORAZIO J, JARRETT S, AMARO-ORTIZ A, et al. UV radiation and the skin[J].International Journal of Molecular Sciences, 2013, 14 (6) :12222-12248.
[9]	[9]JETTE N, LEES-MILLER S P. The DNA-dependent protein kinase:A multifunctional protein kinase with roles in DNA double strand break repair and mitosis[J].Progress in Biophysics and Molecular Biology, 2015, 117 (2-3) :194-205.
[10]	[10]DOUGLAS P, YE R, TRINKLE-MULCAHY L, et al.Polo-like kinase 1 (PLK1) and protein phosphatase 6 (PP6) regulate DNA-dependent protein kinase catalytic subunit (DNA-PKcs) phosphorylation in mitosis[J].Bioscience Reports, 2014, 34 (3) :257-271.
[11]	[11]GOODWIN J F, KOTHARI V, DRAKE J M, et al.DNA-PKcs-Mediated Transcriptional Regulation Drives Prostate Cancer Progression and Metastasis[J].Cancer cell, 2015, 28 (1) :97-113.
[12]	[12]SONE K, PIAO L, NAKAKIDO M, et al. Critical role of lysine 134 methylation on histone H2AX for gamma-H2AX production and DNA repair[J].Nature Communications, 2014, 5 (1) :1-12.
[13]	[13]TU Y, JI C, YANG B, et al. DNA-dependent protein kinase catalytic subunit (DNA-PKcs) -SIN1 association mediates ultraviolet B (UVB) -induced Akt Ser-473 phosphorylation and skin cell survival[J].Molecular Cancer, 2013, 12 (1) :172.
[14]	[14]FLORENCE M E, MASSUDA J Y, SOARES T C, et al.p53immunoexpression in stepwise progression of cutaneous squamous cell carcinoma and correlation with angiogenesis and cellular proliferation[J].Pathology, Research and Practice, 2015, 211 (10) :782-788.

相关文章(20)

[1]	皇甫文丽, 黄瑶, 刘波, 吕长海, 刘娟, 代自超. 建立大鼠颞下颌关节骨关节炎动物模型的2种方法比较, 昆明医科大学学报. doi: 10.12259/j.issn.2095-610X.S20230308
[2]	杨镕羽, 宋飞, 黄浩, 段开文, 钱石兵, 向盈盈. SD大鼠慢性根尖周炎动物模型的研究进展, 昆明医科大学学报. doi: 10.12259/j.issn.2095-610X.S20221005
[3]	闵杰青, 田锦润, 李霁伟, 魏骊铮, 王鹏, 曹彦洁, 何文姬, 樊茂, 张倩, 杨云娟. 二维斑点追踪技术对柔红霉素药物急性期心肌损伤的相关性, 昆明医科大学学报. doi: 10.12259/j.issn.2095-610X.S20220211
[4]	李霁伟, 谢余澄. 内置式双向电极观察先天性巨结肠动物模型肠电图, 昆明医科大学学报.
[5]	赵维佳, 李红宾, 陈宗翰. 窄谱中波紫外线联合卡泊三醇治疗银屑病症状转归及患者治疗前后外周血细胞因子水平评价, 昆明医科大学学报. doi: 10.12259/j.issn.2095-610X.S20201227
[6]	张楠, 胡欣瑜, 董鲜祥, 段为钢, 云宇. 高尿酸血症动物模型的研究进展, 昆明医科大学学报.
[7]	侯云鹤, 张皓鑫, 王依雨, 苏艳梅, 顾丹珊, 林少芳, 杨世昆, 李树德. 人参皂苷Rg1对NAFLD动物模型中同型半胱氨酸的调控机制, 昆明医科大学学报.
[8]	杨灿, 巩宇航, 王海峰, 李海皓, 刘靖宇, 王伟, 王剑松, 左毅刚, 陈戬, 詹辉, 丁明霞. 单抗KMP1的体内抑制膀胱癌EJ细胞株成瘤的动物实验, 昆明医科大学学报.
[9]	任旭, 王沂峰, 戴丽冰. 子宫内膜异位症SCID小鼠动物模型的构建, 昆明医科大学学报.
[10]	秦雨琪, 张丽娟, 任伟东, 董坚. 骨髓干细胞输注修复放疗免疫抑制的动物实验, 昆明医科大学学报.
[11]	张丽娟, 秦雨琪, 任伟东, 董坚. 直接用于输注的临床级细胞冻存保护剂的动物体内外安全性评价, 昆明医科大学学报.
[12]	姜福琼. 重楼皂苷Ⅱ对人皮肤鳞状细胞癌细胞A431增殖的影响, 昆明医科大学学报.
[13]	张浒. 二尖瓣狭窄动物模型的建立, 昆明医科大学学报.
[14]	光雪峰. 左向右分流型肺动脉高压动物模型的制作, 昆明医科大学学报.
[15]	陈刚. 猪胆道缺损修补实验动物模型的建立, 昆明医科大学学报.
[16]	李来邦. 小体积肝移植动物模型的建立, 昆明医科大学学报.
[17]	朱洪. 改进背驮式肝移植动物模型的建立和评价, 昆明医科大学学报.
[18]	. 改进背驮式肝移植动物模型的建立和评价, 昆明医科大学学报.
[19]	. 两种空气消毒方法对心脏导管室接台手术空气消毒效果比较, 昆明医科大学学报.
[20]	王晓莉. DNA-PKcs在乳腺浸润性导管癌中的表达与病理分级的关系, 昆明医科大学学报.