干扰胰岛素样生长因子结合蛋白相关蛋白1的表达对鼻咽癌细胞生物学行为的影响

发布时间：2018-08-29 16:37

【摘要】：目的:采用RNA干扰技术抑制胰岛素样生长因子结合蛋白相关蛋白1(insulin-like growth factor-binding protein related protein 1,IGFBP-rP1)基因表达后,观察鼻咽癌细胞系CNE1和CNE-2Z生物学行为的变化,并探讨其可能的分子机制。方法:通过免疫组织化学法检测25例鼻咽癌组织和22例慢性鼻咽炎组织中IGFBP-rP1蛋白的表达情况。将针对IGFBP-rP1基因的特异性siRNA(同时设置阴性对照)转染鼻咽癌细胞系CNE1和CNE-2Z,然后采用实时荧光定量PCR和蛋白质印迹法鉴定IGFBP-rP1基因的沉默效果,并采用蛋白印迹法检测各组细胞中总的细胞外调节激酶(extracellular regulated kinase,ERK)1/2和磷酸化ERK(phosphorylated ERK,p-ERK)的表达。采用细胞划痕愈合实验、Transwell侵袭实验和CCK-8法分别检测干扰IGFBP-rP1基因表达对鼻咽癌CNE1和CNE-2Z细胞迁移、侵袭和增殖能力的影响。结果:鼻咽癌组织中IGFBP-rP1蛋白的表达明显高于慢性鼻咽炎组织(P0.05)。特异性siRNA转染后,鼻咽癌CNE1和CNE-2Z细胞中IGFBP-rP1基因的表达被明显抑制(P值均0.05),细胞的增殖、迁移和侵袭能力均明显降低(P值均0.05);同时两种细胞中p-ERK表达水平明显下调(P值均0.05),而总ERK1/2的表达水平不变(P值均0.05)。结论:下调IGFBP-rP1基因表达可能通过调控ERK通路,抑制鼻咽癌细胞的增殖、迁移及侵袭。提示IGFBP-rP1有望成为鼻咽癌基因治疗的一个新靶点。
[Abstract]:Aim: to investigate the biological behavior of nasopharyngeal carcinoma (NPC) cell line CNE1 and CNE-2Z after inhibiting the expression of insulin-like growth factor-binding protein-associated protein 1 (insulin-like growth factor-binding protein related protein _ 1IGFBP-rP1) by RNA interference technique, and to explore its possible molecular mechanism. Methods: immunohistochemical method was used to detect the expression of IGFBP-rP1 protein in 25 cases of nasopharyngeal carcinoma and 22 cases of chronic nasopharyngitis. The specific siRNA targeting IGFBP-rP1 gene (with negative control) was transfected into nasopharyngeal carcinoma (NPC) cell lines CNE1 and CNE-2Z,. The silencing effect of IGFBP-rP1 gene was evaluated by real-time fluorescent quantitative PCR and Western blotting. The expression of total extracellular regulated kinase (extracellular regulated kinase,ERK) 1 / 2 and phosphorylated ERK (phosphorylated ERK,p-ERK were detected by Western blot. The effects of interfering IGFBP-rP1 gene expression on the migration, invasion and proliferation of nasopharyngeal carcinoma (NPC) CNE1 and CNE-2Z cells were detected by transwell invasion assay and CCK-8 assay respectively. Results: the expression of IGFBP-rP1 protein in nasopharyngeal carcinoma was significantly higher than that in chronic nasopharyngitis (P0.05). After transfection of specific siRNA, the expression of IGFBP-rP1 gene was significantly inhibited in CNE1 and CNE-2Z cells of nasopharyngeal carcinoma (P < 0. 05). At the same time, the expression of p-ERK was significantly decreased (P < 0. 05), while the expression of total ERK1/2 was not changed (P = 0. 05). Conclusion: down-regulation of IGFBP-rP1 gene expression may inhibit the proliferation, migration and invasion of nasopharyngeal carcinoma cells by regulating ERK pathway. The results suggest that IGFBP-rP1 may be a new target for gene therapy of nasopharyngeal carcinoma.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R739.63

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