宫颈癌细胞GM-CSF与COX-2/iNOS表达的相关性研究

Correlation between GM-CSF and COX-2/iNOS expression in cervical cancer

目的研究宫颈癌细胞中人粒细胞巨噬细胞集落刺激因子(GM-CSF)的表达对环氧合酶-2(COX-2)和诱导性一氧化氮合酶(iNOS)表达的影响,探讨GM-CSF在宫颈癌发生发展中的作用。方法收集临床宫颈癌标本52例及正常宫颈15例和宫颈上皮内瘤变(CIN)组织标本21例作为对照进行免疫组织化学染色,分析宫颈癌组织GM-CSF的表达与COX-2和iNOS表达的相关性;为了从细胞水平进一步论证其相关性,本实验使用宫颈癌细胞系Caski细胞,Caski细胞经GM-CSF转染后过表达GM-CSF,并通过RT-qPCR和Western blot检测COX-2和iNOS mRNA及蛋白的表达情况,以空质粒转染的Caski细胞为对照,在细胞模型中探讨GM-CSF强制表达对COX-2和iNOS表达的影响。同时,通过GM-CSF siRNA干扰Caski细胞GM-CSF表达后,用RTqPCR及Western blot方法检测iNOS/COX-2 mRNA及蛋白表达情况。结果免疫组化结果显示,宫颈癌组织GM-CSF表达明显低于正常组和CIN组(P均<0.05),而COX-2及iNOS的表达显著高于正常组(P<0.01),但与CIN组无明显差异;统计学分析发现GM-CSF与COX-2/iNOS的表达呈负相关(r=-0.542/-0.499)。细胞水平结果显示与空质粒转染的细胞相比,转染GM-CSF的Caski细胞COX-2/iNOS在mRNA及蛋白表达水平都有显著下降(P均<0.05);GM-CSF在被干扰后,干扰组与对照组相比COX-2/iNOS在mRNA及蛋白表达水平有明显差异(P均<0.05)。结论在组织和细胞表达水平GM-CSF与COX-2/iNOS的表达均呈负相关,GM-CSF可能通过下调COX-2/iNOS表达而具有抗宫颈癌作用。

This study performed to explore the roles of GM-CSF in the occurrence and development of cervical cancer by evaluating the effect of GM-CSF expression on COX-2 and iNOS in cervical cancer cells. Firstly, Immunohistochemieal （IHC） assay was used to detect GM-CSF, iNOS and COX-2 in 52 cervical cancer, 21 cervical intraepithelial neoplasia （CIN） and 15 normal cervical samples, for analyzing the correlation between GM- CSF and COX-2/iNOS. To explore whether GM-CSF over-expression had an effect on COX-2/iNOS in cellular models, Caski cells were introduced and transfected with GM-CSF to improve the expression of GM-CSF, furthermore, the mRAN and protein expression of COX-2 and iNOS were detected with RT-qPCR and Western blot, respectively. In control trial, the expression of GM-CSF was silenced in Caski cells to compare the mRNA and protein expression of COX-2/iNOS. IHC results showed GM-CSF in carcinoma group was decreased （P 〈 0.05） as compared with normal tissues and CIN groups, but the COX-2 and iNOS expression were opposite. And the statistic analysis found that the expression of GM-CSF had a negative correlation with COX-2/iNOS（r=-0.542/-0.499）; mRAN expression and protein expression of COX-2 and iNOS were decreased in the cells transfected with GM- CSF（P〈 0.05）, and they were increased when GM-CSF was silenced. The results indicated that GM-CSF may play an anti-cervical carcinoma role by decreasing the expression of COX-2/iNOS.