DNA methylation biomarkers in stool for early screening of colorectal cancer

Jie Chen, Haipeng Sun, Weisen Tang, Lin Zhou, Xi Xie, Zhan Qu, Mengfei Chen, Shunyao Wang, Ting Yang, Ying Dai, Yongli Wang, Tangjie Gao, Qiao Zhou, Zhuo Song, Mingmei Liao, Weidong Liu, Jie Chen, Haipeng Sun, Weisen Tang, Lin Zhou, Xi Xie, Zhan Qu, Mengfei Chen, Shunyao Wang, Ting Yang, Ying Dai, Yongli Wang, Tangjie Gao, Qiao Zhou, Zhuo Song, Mingmei Liao, Weidong Liu

Abstract

Objective: Detection of aberrant methylated genes in feces has been developed as an early screening method for colorectal cancer. The aim of this study was to probe the methylation status of SEPT9, BMP3, NDRG4, and SDC2 in stool and study whether methylation of these genes is associated with colorectal cancer. Materials and Methods: DNAs were isolated and purified from cancerous and non-cancerous stool samples and colorectal cancer tissue. Gene methylation levels were quantified by methylation-specific PCR on SEPT9, BMP3, NDRG4, and SDC2 and analyzed by a diagnostic model. Results: DNA methylation of SEPT9, NDRG4 and SDC2, but not BMP3, had diagnostic potential for detecting colorectal cancer. Moreover, integration of SEPT9, NDRG4, and SDC2 methylation demonstrated high feasibility for detecting colorectal cancer and adenoma, with better performance on colorectal cancer than adenoma. Conclusion: The methylation of SEPT9, NDRG4, and SDC2 in stool may be a potential biomarker for early screening of colorectal cancer.

Keywords: DNA methylation; biomarker; colorectal cancer; stool.

Conflict of interest statement

Competing Interests: The authors have declared that no competing interest exists.

© The author(s).

Figures

Fig 1
Fig 1
Methylation in tissue samples from patients with colorectal cancer. Methylation of four genes in tissue samples. Methylation levels of each gene were measured by qMSP in samples from normal tissue (Norm), non-tumor adjacent tissue (NAT), and tumor tissue (Tumor). (a) Ct values for each gene in different samples. (b) Methylation level represent by ratio to the reference gene ACTB. Data are mean±SD. *P<0.05, **P<0.01, ***P<0.001
Fig 2
Fig 2
Methylation of four genes in stool samples from patients with and without colorectal cancer. Methylation levels of each gene were measured by qMSP in (a) 41 samples of (CRC), 37 of adenoma (Ade), and 152 of normal (Norm) stool. The intersection of each gene detected in (b) adenoma or (c) CRC samples. Data are mean±SD. *P<0.05, ***P<0.001
Fig 3
Fig 3
Diagnostic test with different genes in stool samples. The diagnostic performance with individual genes and the combination of four genes (Combine 4) or three genes, SEPT9, NDRG4, and SDC2 (Combine 3). Receiver operating characteristic (ROC) curves for training cohort (a) and testing cohort (b). (c) Effect of each gene in the Combine 4 model. (d) Comparing the coefficients for the Combine 4 and Combine 3 models.
Fig 4
Fig 4
Diagnostic performance of the Combined 3 model in stool for CRC and adenoma. (a) ROC curves for normal vs adenoma sample (Ade) or CRC sample. (b) AUC and sensitivity for the diagnostic performance in Ade and CRC samples.

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