下调Cyclophilin A对喉鳞癌裸鼠移植瘤模型肿瘤生长和转移的影响

发布时间：2018-03-26 13:43

本文选题：CypA　切入点：慢病毒　出处：《第三军医大学学报》2017年04期

【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(P0.05);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(P0.05)。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。
[Abstract]:Objective to study the effect of cyclin A(Cyclophilin Agna) gene on tumor growth and metastasis in nude mice with laryngeal squamous cell carcinoma (SCC) by RNA interference technique. Methods Lentivirus particles of silencing CypA gene were constructed and transfected into Hep2 laryngeal carcinoma cells. Fifteen nude mice were randomly divided into three groups: blank control group, negative control group and gene silencing group. The expression of CypA, EMMPRIN-CD147, and MMP-9 in tumor tissues was detected by RT-PCR. The expression of CypA, EMMPRIN-CD147 and MMP-9 was detected by RT-PCR. The expression of CypA1CD147 and MMP-9 m in tumor tissues was detected by reverse transcription-polymerase chain reaction (RT-PCR). The expression of MMP-9 m in tumor tissues was detected by reverse transcription-polymerase chain reaction (RT-PCR). The expression of MMP-9 protein was detected by Western blot. Results the tumor volume in the gene silencing group was higher than that in the blank control group. In the negative control group, the difference was statistically significant (P 0.05), and the expression of CypA was mainly expressed in the cytoplasm, CD147, and the expression of MMP-9 was mainly expressed in the stroma. The expression of MMP-9 in CypAfir CD147m9 was significantly decreased in the group of gene silencing of m RNA level and protein level. Conclusion down-regulation of CypA can inhibit the growth of laryngeal carcinoma and delay the metastasis of laryngeal carcinoma by regulating the expression level of CD147 MMP 9.
【作者单位】：重庆医科大学附属第一医院耳鼻咽喉科;
【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)~~
【分类号】：R739.65;R-332

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