Real-time colorimetric detection of DNA methylation of the &lt;em&gt;PAX1&lt;/em&gt; gene in cervical scrapings for cervical cancer screening with thiol-labeled PCR primers and gold nanoparticles

Huang, Jin; Liou, Yu-Ligh; Kang, Yi Lan; Tan, Zhi‐Rong; Zhou, Hong‐Hao

doi:10.2147/ijn.s116288

Cited by 17 publications

(14 citation statements)

References 42 publications

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“…GSEA results further confirmed the identified pathways and simultaneously found some new biological processes DEGs significantly enriched such as RNA splicing and Gene silencing 72, 73. Mole S, et al demonstrated that HPV16 E2 as a transcription factor could regulate the expression of RNA splicing factors SF2/ASF, SRp20 and SC35 to facilitate cervical cancer progression 74.…”

Section: Discussionmentioning

confidence: 76%

Identification of Key Genes and Pathways in Cervical Cancer by Bioinformatics Analysis

Wu¹,

Peng²,

Zhang³

et al. 2019

Int. J. Med. Sci.

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Cervical cancer is a common malignant tumour of the female reproductive system that seriously threatens the health of women. The aims of this study were to identify key genes and pathways and to illuminate new molecular mechanisms underlying cervical cancer. Altogether, 1829 DEGs were identified, including 794 significantly down-regulated DEGs and 1035 significantly up-regulated DEGs. GO analysis suggested that the up-regulated DEGs were mainly enriched in mitotic cell cycle processes, including DNA replication, organelle fission, chromosome segregation and cell cycle phase transition, and that the down-regulated DEGs were primarily enriched in development and differentiation processes, such as tissue development, epidermis development, skin development, keratinocyte differentiation, epidermal cell differentiation and epithelial cell differentiation. KEGG pathway analysis showed that the DEGs were significantly enriched in cell cycle, DNA replication, the p53 signalling pathway, pathways in cancer and oocyte meiosis. The top 9 hub genes with a high degree of connectivity (over 72 in the PPI network) were down-regulated TSPO, CCND1, and FOS and up-regulated CDK1, TOP2A, CCNB1, PCNA, BIRC5 and MAD2L1. Module analysis indicated that the top 3 modules were significantly enriched in mitotic cell cycle, DNA replication and regulation of cell cycle ( P < 0.01). The heat map based on TCGA database preliminarily demonstrated the expression change of the key genes in cervical cancer. GSEA results were basically coincident with the front enrichment analysis results. By comprehensive analysis, we confirmed that cell cycle was a key biological process and a critical driver in cervical cancer. In conclusion, this study identified DEGs and screened the key genes and pathways closely related to cervical cancer by bioinformatics analysis, simultaneously deepening our understanding of the molecular mechanisms underlying the occurrence and progression of cervical cancer. These results might hold promise for finding potential therapeutic targets of cervical cancer.

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Section: Discussionmentioning

confidence: 76%

Identification of Key Genes and Pathways in Cervical Cancer by Bioinformatics Analysis

Wu¹,

Peng²,

Zhang³

et al. 2019

Int. J. Med. Sci.

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“…One is GynTect ® produced by Oncgnostics company of German and another is QIAsure ® by QIAGEN and Self-screen BV company. According to the product specifications for GynTect ® 22,23 The cycle threshold (Ct) values for each target were normalized for DNA input. The values for all samples were transformed into relative quantity.…”

Section: Discussionmentioning

confidence: 99%

<p>A Sensitive and Simplified Classifier of Cervical Lesions Based on a Methylation-Specific PCR Assay: A Chinese Cohort Study</p>

Zhang¹,

Yu²,

Huang³

et al. 2020

CMAR

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These authors contributed equally to this workObjective: The aim of this study is to assess the diagnostic and screening performance of a standardized methylation-specific real-time PCR assay targeting SOX1 and PAX1 genes for cervical cancer in a Chinese cohort. Methods: Genomic DNA was extracted from cervical exfoliated cells and converted by sodium bisulfite and then analyzed by qMSP assay. Ct values were collected for PAX1 and SOX1 as target genes and β-actin as an endogenous reference gene. The samples included 295 cervicitis, 111 LSIL (low-grade squamous intraepithelial lesion), 51 HSIL (high-grade squamous intraepithelial lesion) and 30 cervical cancer. Results: The Ct values decreased with the progression of cervical cancer from cervicitis, through LSIL and HSIL to cancer. The difference in Ct values between cytological grades was highly significant (p≤0.01) between grades either for PAX1 or for SOX1 except the difference between cervicitis and LSIL of SOX1. With the Ct cut-off values of PAX1 gene and SOX1 gene 38.6 and 38 and with the PAX1/SOX1 in combination, the positive rate of methylation in invasive cancer tissues was 100%, in contrast to 11.5% (95% CI: 8.67%-14.33%) in cervicitis tissues, 45.1% (95% CI: 40.68%-49.52%) in LSIL tissues, and 68.5% (95% CI: 64.37%-72.63%) in HSIL tissues. The specificity and sensitivity of differentiating tumors from cervicitis were 0.957 (95% CI: 0.939-0.975) and 1.00, respectively. The specificity and sensitivity of differentiation between cervicitis+LSIL and HSIL+cervical cancer were 0.881 (95% CI: 0.852-0.91) and 0.748 (95% CI: 0.709-0.787), respectively. Conclusion: PAX1/SOX1 methylation could be translated into clinical practice for cervical neoplasia detection.

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“…The methylation of APC1A promoter was also closely correlated with biological features in CC and could therefore be used to predict poor prognosis of CC [34]. DNA methylation could result in carcinogenesis through silencing important tumor suppressor genes and aberrant methylation of tumor suppressor genes could be regarded as a prognostic and predictive biomarker for cancer [35]. An increase in the promotor methylation of Ezrin in CC cases would normally cause a decrease in gene transcription.…”

Section: Discussionmentioning

confidence: 99%

Promoter Methylation of Ezrin and its Impact on the Incidence and Prognosis of Cervical Cancer

Qin¹,

Cao²,

Wang³

et al. 2018

Cell Physiol Biochem

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Background/Aims: Aberrant localization and over-expression of Ezrin have been reported to be implicated in cervical cancer (CC). Aberrant promoter methylation of some gene families may serve as potential diagnostic biomarkers for CC. In this study, we explored the correlation of promoter methylation of the Ezrin gene with the incidence and prognosis of CC. Methods: Cervical tissues from a total of 483 patients with CC were collected from the China-Japan Union Hospital of Jilin University. Samples were assigned into four groups accordingly to pathological diagnosis, namely the control group, the cervical intraepithelial neoplasia (CIN) I group, the CIN II-III group and the CC group. Reverse transcription quantitative polymerase chain reaction (RT-qPCR) was performed to detect the mRNA expression of Ezrin. Methylation-specific polymerase chain reaction (MSP) was used to detect the promoter methylation of the Ezrin gene. The Kaplan-Meier product-limit method and the log-rank analysis were used for survival analysis, the Cox regression analysis for the prognostic factors for CC, and the logistic regression analysis for the risk factors for the occurrence of CC. Results: The methylation rate of the Ezrin gene was correspondingly increased from the control, the CIN I, the CIN II-III to the CC groups. Over-expressed mRNA of Ezrin was determined in CC tissues. The mRNA expression of Ezrin was correlated with tumor size, lymphatic metastasis, pathological grade and clinical stage (FIGO). The risk factors for the occurrence of CC were the number of abortions and the promoter methylation of the Ezrin gene. Poor prognosis of CC correlated to lymphatic metastasis, higher pathological grade, higher FIGO stage and positive Ezrin promoter methylation. Conclusion: These findings indicate that promoter methylation of the Ezrin gene may play a crucial role in carcinogenesis, progression and prognosis of CC.

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Real-time colorimetric detection of DNA methylation of the <em>PAX1</em> gene in cervical scrapings for cervical cancer screening with thiol-labeled PCR primers and gold nanoparticles

Cited by 17 publications

References 42 publications

Identification of Key Genes and Pathways in Cervical Cancer by Bioinformatics Analysis

Identification of Key Genes and Pathways in Cervical Cancer by Bioinformatics Analysis

<p>A Sensitive and Simplified Classifier of Cervical Lesions Based on a Methylation-Specific PCR Assay: A Chinese Cohort Study</p>

Promoter Methylation of Ezrin and its Impact on the Incidence and Prognosis of Cervical Cancer

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