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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 study the effect of Cyclophilin A (CypA) gene on tumor growth and tumor metastasis in nude mice with laryngeal squamous cell carcinoma by RNAi technique. Methods The lentiviral vector containing CypA gene was 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, with 5 in each group. Transplantation model of xenografts in nude mice was observed. The growth of tumor and lymph node metastasis were observed. The expression of CypA, extracellular matrix metalloproteinase inducer (EMMPRIN, CD147) and extracellular matrix metalloproteinase-9 (MMP-9) in the tumor tissue were detected by PV two-step immunohistochemistry. -9 m RNA expression levels, Western blot was detected CypA, CD147, MMP-9 protein expression levels. Results The tumor volume of gene silencing group was smaller than that of blank control group and negative control group (P <0.05). Immunohistochemistry showed that the tumor was moderately differentiated squamous cell carcinoma, CypA mainly expressed in cytoplasm, and CD147 mainly expressed in The expression of CypA, CD147 and MMP-9 in the cell membrane and MMP-9 was mainly in the stroma of tumor. The levels of CypA, CD147 and MMP-9 were significantly decreased in the m RNA level and protein level gene silencing group (P <0.05). Conclusion The down-regulation of CypA can inhibit the growth of laryngeal carcinoma by regulating the expression of CD147 and MMP-9 and delay the metastasis.