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-rP 1)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-r P1蛋白的表达情况。将针对IGFBP-r P1基因的特异性si RNA(即IGFBP-r P1 si RNA)及其阴性对照si RNA分别转染鼻咽癌CNE1和CNE-2Z细胞,然后采用实时荧光定量PCR和蛋白质印迹法鉴定IGFBPr P1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)和磷酸化ERK(phosphorylated ERK,p-ERK)的表达。采用CCK-8法、细胞划痕愈合实验和Transwell小室法分别检测干扰IGFBP-r P1基因表达对鼻咽癌CNE1和CNE-2Z细胞增殖、迁移和侵袭能力的影响。结果 :鼻咽癌组织中IGFBP-r P1蛋白的表达水平明显高于慢性鼻咽炎组织(P<0.05)。IGFBP-r P1 si RNA转染后,鼻咽癌CNE1和CNE-2Z细胞中IGFBP-r P1基因的表达均被明显抑制(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2种细胞中p-ERK表达水平明显下调(P值均<0.01),而总ERK的表达水平不变(P值均>0.05)。结论 :下调IGFBP-r P1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-r P1有望成为鼻咽癌基因治疗的一个新靶点。
Objective： To observe the changes of biological behaviors of nasopharyngeal carcinoma cell lines CNE1 and CNE-2Z after the expression of insulin-like growth factor-binding protein related protein 1 （IGFBP-rP1） was inhibited by RNA interference, and to explore the possible molecular mechanism. Methods： The expression of IGFBP-rP1 in 25 cases of nasopharyngeal carcinoma tissues and 22 cases of chronic nasopharyngitis tissues was measured by immunohistochemical method. The specific siRNAs targeting ICFBP-rP1 gene （IGFBP-rP1 siRNAs） and the negative control siRNA were transfected into nasopharyngeal carcinoma cell lines CNE1 and CNE-2Z, respectively. Then the efficiency of IGFBP-rP1 gene-siliencing was verified by real-time fluorescent quantitative PCR and Western blotting. Meanwhile, the expressions of total extracellular regulated kinase （ERK） and phosphorylated ERK （p-ERK） were detected by Western blotting. Then the proliferation, migration and invasion of CNE1 and CNE-2Z cells transfected with IGFBP-rP1 siRNAs were examined by CCK-8 assay, wound healing assay and Transwell invasion assay, respectively. Results： The expression level of IGFBP-rP1 protein in nasopharyngeal carcinoma tissues was higher than that in chronic nasopharyngitis tissues （P 〈 0.05）. After transfection with IGFBP-rP1 siRNAs, the expressions of IGFBP-rP1 mRNA and protein in CNE1 and CNE- 2Z cells were significantly down-regulated （all P 〈 0.05）; the cell proliferation, migration and invasion of CNE1 and CNE-2Z cells were significantly inhibited （all P 〈 0.05）; the expression level of p-ERK was significantly decreased （both P 〈 0.01）, while the expression level of total ERK was unchanged in CNE1 and CNE-2Z cells （both P 〉 0.05）. Conclusion： Down-regulation of IGFBP-rP1 gene expression maybe inhibit the proliferation, migration and invasion of nasopharyngeal carcinoma cells by regulating the ERK pathway. It is suggested that IGFBP-rP1 probably becomes a new target for the gene therapy of nasopharyngeal carcinoma.
出处
《肿瘤》
CAS
CSCD
北大核心
2017年第1期33-40,82,共9页
Tumor
基金
重庆市永川区2013年科技计划项目(编号:Ycstc
2013nc8014)
重庆医科大学附属永川医院2013年度院内科研课题(编号:YJYB201309)~~
