Department of Oncology, The Second Affiliated Hospital of Soochow University, Suzhou, 215004, China.
Department of Pharmacy, The Affiliated Children's Hospital of Soochow University, Suzhou, 215004, China.
Appl Biochem Biotechnol. 2023 Dec;195(12):7821-7831. doi: 10.1007/s12010-023-04488-9. Epub 2023 Apr 24.
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.
PIK3CA 突变被认为是治疗结直肠癌的潜在靶点。我们评估了一种新开发的聚合酶链反应（PCR）联合限制酶消化和 Sanger 测序，用于检测血浆无细胞游离 DNA（cfDNA）中的 PIK3CA 突变。我们使用新开发的检测方法分析了血浆中的 PIK3CA 突变，并通过常规方法分析了匹配的肿瘤组织中的 PIK3CA 突变。我们使用这两种方法检测了 40 例结直肠癌患者的样本中的 PIK3CA 基因突变状态。通过限制酶消化 PCR 在 40 例患者的 cfDNA 中检测到 3 个 PIK3CA 基因的 H1047R 突变。常规 PCR/测序未检测到 E545K 或 H1047R 突变。我们新开发的检测方法可以敏感地检测到 cfDNA 中的 PIK3CA H1047R 和 E545K 突变。结直肠癌已被用作检测我们新检测方法的临床实例，这表明新的检测方法可能在精准肿瘤学的突变检测中有更广泛的应用。试验注册：当前对照试验 ChiCTR-DDT-12002848，2012 年 10 月 8 日。
