Optimization to detect TP53 mutations in circulating cell-free tumor DNA from patients with serous epithelial ovarian cancer.
Optimization to detect TP53 mutations in circulating cell-free tumor DNA from patients with serous epithelial ovarian cancer.
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DOI：
10.5468/ogs.2018.61.3.328
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发表时间：
2018-05
影响因子：
--
通讯作者：
Kim YT
中科院分区：
文献类型：
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作者：
Park YR;Kim YM;Lee SW;Lee HY;Lee GE;Lee JE;Kim YT
循环无细胞肿瘤DNA（cfDNA）是在肿瘤发展期间由原发性肿瘤的凋亡和坏死细胞释放到血液中的DNA。cfDNA反映了原始肿瘤的遗传和表观遗传改变。TP 53突变是高级别浆液性卵巢癌的一个定义性特征。我们优化了用于检测来自血液样品的cfDNA中TP 53突变的方法。我们使用数字聚合酶链反应（dPCR）证实了原发性卵巢癌组织中TP 53突变与cfDNA中TP 53突变的相关性。我们利用癌症基因组图谱和癌症体细胞突变目录的数据，在TP 53中发现了12个常见的突变位点，并制备了12条引物。在新鲜冷冻组织（FFT）和福尔马林固定石蜡包埋组织（FFPET）中评估组织中的突变。我们对4例患者在减积手术前采集的系列血浆样本进行了前瞻性分析。我们提取cfDNA并计算其在血液中的浓度。采用dPCR方法检测cfDNA中TP 53突变，并比较卵巢癌组织与cfDNA中TP 53突变。12个引物中的10个检测到FFT、FFPET和cfDNA中TP 53突变的存在。在FFT和FFPET组织中，没有显著差异。平均cfDNA浓度为2.12±0.59 ng/mL。我们还证实了cfDNA和FFT的突变都在R282 W位点。本研究开发了使用dPCR检测卵巢癌患者cfDNA中TP 53突变的方法。结果表明，在卵巢癌组织和cfDNA中存在相同的TP 53突变。
Circulating cell-free tumor DNA (cfDNA) is the DNA released by apoptotic and necrotic cells of the primary tumor into the blood during the period of tumor development. The cfDNA reflects the genetic and epigenetic alterations of the original tumor. TP53 mutations are a defining feature of high-grade serous ovarian carcinoma. We optimized the methods for detecting TP53 mutations in cfDNA from blood samples. We confirmed the correlation of TP53 mutation in primary ovarian cancer tissue and it in cfDNA using digital polymerase chain reaction (dPCR). We found 12 frequent mutation sites in TP53 using The Cancer Genome Atlas and Catalogue of Somatic Mutations in Cancer data and manufactured 12 primers. The mutations in tissues were evaluated in fresh-frozen tissue (FFT) and formalin-fixed paraffin-embedded tissue (FFPET). We performed a prospective analysis of serial plasma samples collected from 4 patients before debulking surgery. We extracted cfDNA and calculated its concentration in blood. dPCR was used to analyze TP53 mutations in cfDNA, and we compared TP53 mutations in ovarian cancer tissue with those in cfDNA. Ten primers out of 12 detected the presence of TP53 mutations in FFT, FFPET, and cfDNA. In FFT and FFPET tissue, there were no significant differences. The average cfDNA concentration was 2.12±0.59 ng/mL. We also confirmed that mutations of cfDNA and those of FFT were all in R282W site. This study developed detection methods for TP53 mutations in cfDNA in ovarian cancer patients using dPCR. The results demonstrated that there are the same TP53 mutations in both ovarian cancer tissue and cfDNA.