摘要
目的通过RNA干扰技术,研究亲环蛋白A(Cyclophilin A,CypA)基因对喉鳞癌裸鼠移植瘤模型在肿瘤生长和肿瘤转移方面的影响。方法构建沉默CypA基因慢病毒颗粒,转染Hep2喉癌细胞,将15只裸鼠完全随机分为3组,分别为空白对照组、阴性对照组、基因沉默组,每组5只,建立喉鳞癌裸鼠移植瘤转移模型,观察肿瘤生长情况及淋巴结转移情况。PV二步法免疫组化检测CypA、细胞外基质金属蛋白酶诱导因子(EMMPRIN、CD147)、细胞外基质金属蛋白酶-9(MMP-9)在肿瘤组织的表达,RT-PCR检测CypA、CD147、MMP-9 m RNA的表达水平,Western blot法检测CypA、CD147、MMP-9蛋白的表达水平。结果基因沉默组肿瘤体积较空白对照组、阴性对照组小,差异有统计学意义(P<0.05);免疫组化观察肿瘤为中分化鳞状细胞癌,CypA主要表达在细胞质,CD147主要表达在细胞膜,MMP-9主要表达在肿瘤间质;在m RNA水平和蛋白水平基因沉默组CypA、CD147、MMP-9的表达都明显降低(P<0.05)。结论下调CypA可以通过调节CD147、MMP-9的表达水平抑制喉癌的生长,延缓肿瘤转移。
Objective To explore the effect of RNA interference-mediated cyclophilin A（ CypA） downregulation on the growth and metastasis of laryngeal squamous cell carcinoma in nude mice. Methods Lentiviral vector containing CypA sh RNA was constructed and trans infected into Hep2 cells. Fifteen nude mice were randomly divided into 3 groups（ n = 5） ： control group,NC sh RNA group,and CypA sh RNA group. The growth and lymph node metastasis of tumors were observed. The expression of CypA,CD147 and matrix metalloproteinase（ MMP）-9 in tumor tissues was detected by immunohistochemistry（ IHC）. The m RNA and protein expression levels of CypA,CD147 and MMP-9 were detected by real-time polymerase chain reaction（ RT-PCR） assay and Western blotting,respectively. Results The gross tumor volume of CypA sh RNA group was significantly smaller than those of both control and NC sh RNA groups（ P〈0. 05）. IHC staining showed cytoplasmic localization of CypA expression,whereas expression of CD147 and MMP-9 was detected in the cytomembrane and tumor stroma,respectively. The m RNA and protein expression levels of CypA,CD147 and MMP-9 were significantly decreased in CypA sh RNA group as compared to control and NC sh RNA groups（ P〈0. 05）. Conclusion Down-regulation of cyclophilin A can inhibit the growth and metastasis of laryngeal squamous cell carcinoma by mediating the expression of CD147 and MMP-9.
作者
柏露
周建荣
王致娟
刘舟
Bai Lu Zhou Jianrong Wang Zhijuan Liu Zhou(Department of Otorhinolaryngology, the First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, China)
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2017年第4期355-360,共6页
Journal of Third Military Medical University
基金
国家临床重点专科建设项目(卫办医政函[2012]649号)
重庆市卫生局医学科研重点项目(20121024)~~
关键词
CYPA
慢病毒
RNA干扰
喉癌
裸鼠
cyclophilin A
lentiviral vector
RNA interference
laryngeal squamous cell carcinoma
nude mice
