Abstract
The PIK3CA mutation is considered a potential target for treatment of colorectal cancer. We evaluated a PIK3CA mutation assay on plasma cell-free DNA (cfDNA) using a newly developed PCR with restriction digestion integrated and followed by Sanger’s sequencing. We analyzed PIK3CA mutation in plasma with our newly developed assays and in matching tumor tissues by routine methods. We detected the PIK3CA gene mutation status by both methods in samples from 40 colorectal cancer patients. Three H1047R mutations of PIK3CA gene were detected in the cfDNA of the 40 patients by restriction digestion PCR. Neither E545K nor H1047R mutations were detected in the cfDNA by routine PCR/sequencing. The PIK3CA H1047R and E545K mutations in cfDNA can be sensitively detected with our newly developed assays. The colorectal cancer has been used as a clinical example in testing our new assays, which indicates that the new assays may have wider applications in detecting mutations in precision oncology. Trial registration: Current Controlled Trials ChiCTR-DDT-12002848, 8 October 2012.
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Funding
We acknowledge the Suzhou Science and Technology Planning Project (SKY2021047), the Program of Clinical Research Center of Neurological Disease (ND2022B04), and the Project of State Key Laboratory of Radiation Medicine and Protection, Soochow University (GZK1202024, GZK1202234).
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The study conception and design were contributed by Kai Li and Li Xiao. Material preparation, data collection, and analysis were performed by Fengjiao Wang, Hui Wang, and Huahui Bian. The first draft of the manuscript was written by Wangyang Pu and Chungen Xing. All authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.
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The study has been approved by the human research ethics committee of the Second Affiliated Hospital of Soochow University. Preoperative informed consent was obtained from each patient registered in this study in accordance with institutional guidance. Blood samples were drawn before antitumor therapy and at selected time points following operation, which would not result in any disadvantages to health and prognosis of patients. We are committed to maintain the privacy of patients’ information.
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Pu, W., Wang, F., Li, K. et al. Novel Method for Detection of PIK3CA Mutations in Circulating Tumor DNA of Patients with Colorectal Cancer. Appl Biochem Biotechnol 195, 7821–7831 (2023). https://doi.org/10.1007/s12010-023-04488-9
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DOI: https://doi.org/10.1007/s12010-023-04488-9