He Yunan, Hu Shunjie, Zhong Jiaojiao, Cheng Anran, Shan Nianchun
Department of Gynecology and Obstetrics, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, Guangdong, China.
Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, Guangdong, China.
PeerJ. 2020 Dec 2;8:e10386. doi: 10.7717/peerj.10386. eCollection 2020.
Cervical squamous cancer (CESC) is an intractable gynecological malignancy because of its high mortality rate and difficulty in early diagnosis. Several biomarkers have been found to predict the prognose of CESC using bioinformatics methods, but they still lack clinical effectiveness. Most of the existing bioinformatic studies only focus on the changes of oncogenes but neglect the differences on the protein level and molecular biology validation are rarely conducted.
Gene set data from the NCBI-GEO database were used in this study to compare the differences of gene and protein levels between normal and cancer tissues through significant pathway selection and core gene signature analysis to screen potential clinical biomarkers of CESC. Subsequently, the molecular and protein levels of clinical samples were verified by quantitative transcription PCR, western blot and immunohistochemistry.
Three differentially expressed genes (RFC4, MCM2, TOP2A) were found to have a significant survival ( < 0.05) and highly expressed in CESC tissues. Molecular biological verification using quantitative reverse transcribed PCR, western blotting and immunohistochemistry assays exhibited significant differences in the expression of RFC4 between CESC and para-cancerous tissues ( < 0.05).
This study identified three potential biomarkers (RFC4, MCM2, TOP2A) of CESC which may be useful to clarify the underlying mechanisms of CESC and predict the prognosis of CESC patients.
宫颈鳞状细胞癌(CESC)是一种难治性妇科恶性肿瘤,因其死亡率高且早期诊断困难。已经发现几种生物标志物可使用生物信息学方法预测CESC的预后,但它们仍缺乏临床有效性。大多数现有的生物信息学研究仅关注癌基因的变化,而忽略了蛋白质水平的差异,并且很少进行分子生物学验证。
本研究使用来自NCBI-GEO数据库的基因集数据,通过显著通路选择和核心基因特征分析来比较正常组织和癌组织之间基因和蛋白质水平的差异,以筛选CESC的潜在临床生物标志物。随后,通过定量转录PCR、蛋白质免疫印迹和免疫组织化学验证临床样本的分子和蛋白质水平。
发现三个差异表达基因(RFC4、MCM2、TOP2A)具有显著的生存意义(P<0.05),且在CESC组织中高表达。使用定量逆转录PCR、蛋白质免疫印迹和免疫组织化学分析进行的分子生物学验证显示,CESC与癌旁组织之间RFC4的表达存在显著差异(P<0.05)。
本研究鉴定了CESC的三个潜在生物标志物(RFC4、MCM2、TOP2A),这可能有助于阐明CESC的潜在机制并预测CESC患者的预后。