KCNQ5 and C9orf50 Methylation in Stool DNA for Early Detection of Colorectal Cancer

Yaping Cao, Guodong Zhao, Mufa Yuan, Xiaoyu Liu, Yong Ma, Yang Cao, Bei Miao, Shuyan Zhao, Danning Li, Shangmin Xiong, Minxue Zheng, Sujuan Fei, Yaping Cao, Guodong Zhao, Mufa Yuan, Xiaoyu Liu, Yong Ma, Yang Cao, Bei Miao, Shuyan Zhao, Danning Li, Shangmin Xiong, Minxue Zheng, Sujuan Fei

Abstract

Background: Aberrant DNA methylation has emerged as a class of promising biomarkers for early colorectal cancer (CRC) detection, but the performance of methylated C9orf50 and methylated KCNQ5 in stool DNA has never been evaluated.

Methods: Methylation specific quantitative PCR (qPCR) assays for methylated C9orf50 and methylated KCNQ5 were developed. The methylation levels of C9orf50 and KCNQ5 in 198 CRC patients, 20 advanced adenoma (AA) patients, 101 small polyp (SP) patients, and 141 no evidence of disease (NED) subjects were analyzed.

Results: The methylation levels of both KCNQ5 and C9orf50 genes were significantly higher in CRC and AA groups than those in SP and NED groups, but showed no significant difference among different stages of CRC. The sensitivities of methylated KCNQ5 and methylated C9orf50 for CRC detection were 77.3% (95% CI: 70.7-82.8%) and 85.9% (95% CI: 80.0-90.2%) with specificities of 91.5% (95% CI: 85.3-95.3%) and 95.0% (95% CI: 89.7-97.8%), respectively. When C9orf50 and methylated KCNQ5 were combined, the clinical performance for CRC detection was similar to that of methylated C9orf50 alone.

Conclusions: Stool DNA based methylated C9orf50 test has the potential to become an alternative approach for CRC screening and prevention.

Keywords: colorectal cancer; early detection; methylated C9orf50; methylated KCNQ5; stool DNA.

Conflict of interest statement

Authors GZ and SX were employed by Suzhou VersaBio Technologies Co. Ltd. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Copyright © 2021 Cao, Zhao, Yuan, Liu, Ma, Cao, Miao, Zhao, Li, Xiong, Zheng and Fei.

Figures

Figure 1
Figure 1
The flowchart of this study.
Figure 2
Figure 2
The methylation levels of KCNQ5 and C9orf50 in no evidence of disease (NED), small polyp (SP), advanced adenoma (AA), colorectal cancer (CRC) groups (A, B) and different stages of CRC (C, D). **p < 0.01; ***p < 0.0001, ns, no significant difference according to Student’s t-test. Red lines represent the median methylation levels of KCNQ5 or C9orf50. Mann–Whitney U test compared to methylation levels of KCNQ5 and C9orf50 in NED, SP, AA, CRC groups.
Figure 3
Figure 3
Sensitivities and specificities of methylated KCNQ5 and methylated C9orf50 in stool DNA for small polyp (SP), advanced adenoma (AA), and colorectal cancer (CRC) detection (A), and the receiver operating characteristic (ROC) curves of methylated KCNQ5 and methylated C9orf50 in stool DNA for CRC detection (B).

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