极光激酶A在宫颈鳞状细胞癌患者血清中的表达及其临床意义

目的:应用生物信息学技术分析和筛选影响宫颈癌(CC)预后的关键基因并探讨极光激酶A(AURKA)在宫颈鳞状细胞癌(CSCC)中的表达及其临床意义。方法:从基因表达综合(GEO)数据库下载并分析CC患者的mRNA表达谱及临床数据,利用生物信息学技术筛选出CC组织与正常宫颈组织间的差异表达基因(DEGs)。PPI网络和生存分析结果显示AURKA基因的表达和CC分期及患者预后相关。利用GEPIA2、HPA数据库分别验证AURKA mRNA及蛋白在正常宫颈组织和不同病理分期CC组织中的表达情况。采用Kaplan-Meier法分析CC患者的AURKA基因高、低表达(以中位数为截断值)与生存期是否相关。随后进一步通过实时荧光定量PCR(qPCR)和酶联免疫吸附试验(ELISA)技术分别检测在CSCC、宫颈上皮内瘤变(CIN)和女性健康体检者血清中AURKA mRNA及蛋白的表达。结果:GEPIA2数据库分析结果显示AURKA mRNA在CC组织中高表达(P<0.05),且AURKA mRNA在CC不同阶段的表达水平存在显著差异(P<0.05),随着肿瘤分期的进展AURKA mRNA的表达升高。HPA数据库分析结果显示,与正常宫颈组织相比,AURKA蛋白表达水平在CC组织中亦升高,差异具有统计学意义(P<0.05)。Kaplan-Meier生存分析结果显示,AURKA基因高表达的CC患者生存时间更短。qPCR和ELISA实验显示,CSCC和CIN患者血清AURKA mRNA的相对表达水平明显高于健康体检者(P<0.05)。CSCC患者血清AURKA蛋白的表达水平明显高于CIN及健康体检者(P<0.05)。AURKA mRNA及蛋白在CC血清中均高表达。ROC曲线及χ^(2)检验分析结果显示AURKA mRNA的诊断价值较高,血清AURKA mRNA高表达组患者FIGO分期、淋巴结转移、肿瘤体积及肌层浸润深度均分别高于AURKA mRNA低表达组(P<0.05)。结论:CSCC患者血清中AURKA mRNA及蛋白的表达水平升高,两者联合检测对CSCC的诊断及预后监测具有潜在应用价值。

Expression and clinical significance of AURKA in cervical cancer

OBJECTIVE:To screen for key genes affecting prognosis of cervical cancer (CC),and to explore expression and prognostic values of AURKA in cervical squamous cell carcinomas (CSCC). METHODS:mRNA expression profiles and clinical data from GEO database of CC patients were downloaded,and the differentially expressed genes (DEGs) between cervical and normal cervical tissues were screened by bioinformatics technology. PPI network and survival analyses showed that AURKA was a potential biomarker associated with CC staging and prognosis. GEPIA2 and HPA databases were used to verify expressions of AURKA mRNA and its encoded protein in both cervical tissues,and in different stages of cancer. Kaplan-meier method was used to analyze whether the high and low expression of AURKA (median truncated value) was significantly correlated with survival in patients with cervical cancer. Subsequently,quantitative real-time PCR (qPCR) and Enzyme-linked immunosorbent assay (ELISA) were used to detect the mRNA and protein levels of AURKA in serum of CSCC patients,cervical intraepithelial neoplasia patients and healthy female subjects. RESULTS:GEPIA2 database analyses showed high expression of AURKA mRNA in cervical cancer tissues (P<0.05). Further analysis of the relationship with cervical cancer stages showed that the expression levels in different cancer stages were significantly different (P<0.05),and the levels increased with the progression of tumor staging. Compared with normal cervical tissues,expression of the AURKA protein was also increased using the HPA database analyses,and the difference was statistically significant (P<0.05). Kaplan-Meier survival analyses showed that patients with higher AURKA gene expressions had shorter survival times. qPCR and ELISA showed that the relative expressions of AURKA mRNA in CSCC patients and cervical intraepithelial neoplasia were significantly higher than that in healthy subjects,the difference was statistically significant (P<0.05). Expressions of serum AURKA protein in CSCC patients was significantly higher than that in patients with cervical intraepithelial neoplasia and health subjects (P<0.05). Results from ROC curve and χ² test showed that the diagnostic value of AURKA mRNA was high. The FIGO staging,lymph node metastasis,tumor size and muscle layer infiltration depth of the serum AURKA mRNA high expression group were higher than those in the AURKA mRNA low expression group (P<0.05). CONCLUSION:Expressions of AURKA mRNA and protein in the serum of CSCC patients were elevated,and the combination of the two tests had potential application value for the diagnosis of CSCC and the monitoring of prognosis.