| Mingqiang Kang,Xiangqi Chen,Tingyan Lin,zhisen gao,Jiangjiang Xu,Rui Feng,Sheng Yang,Shuchen Chen. Quantitative monitoring and mutations of ctDNA before and after non-small cell lung cancer radical surgery. Oncol Transl Med, 2019, 5: 103-108. |
| 非小细胞肺癌根治术前后ctDNA定量监测及突变情况的初步研究 |
| Quantitative monitoring and mutations of ctDNA before and after non-small cell lung cancer radical surgery |
| Received:February 20, 2019 Revised:July 23, 2019 |
| DOI：10.1007/s10330-019-0335-5 |
| 中文关键词: 循环肿瘤DNA，非小细胞肺癌，基因突变，分子生物学标志物 |
| 英文关键词: ctDNA; non-small cell lung cancer (NSCLC); mutant genes; molecular markers |
| 基金项目: |
| Author Name | Affiliation | E-mail | | Mingqiang Kang* | Fujian Medical University Union Hospital | drkang2011@163.com | | Xiangqi Chen | Fujian Medical University Union Hospital | | | Tingyan Lin | Fujian Medical University Union Hospital | | | zhisen gao | Fujian Medical University Union Hospital | | | Jiangjiang Xu | Fujian Medical University Union Hospital | | | Rui Feng | Fujian Medical University Union Hospital | | | Sheng Yang | Fujian Medical University Union Hospital | | | Shuchen Chen | Fujian Medical University Union Hospital | |
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| 中文摘要: |
| 目的：研究外周游离循环肿瘤DNA（circulating tumour DNA, ctDNA）在非小细胞肺癌（non-small cell lung carcinoma, NSCLC）患者接受根治性手术治疗前后的定量表达情况，并探讨非小细胞肺癌组织常见突变基因的突变情况与ctDNA中检测到的突变基因的关系。方法：我们随机选取福建医科大学附属协和医院胸外科5例接受根治性手术治疗的非小细胞肺癌患者。① 采集患者手术前以及手术后21-37天静脉血液样本检测ctDNA。采集样本分离出血浆与血细胞，应用QIAamp Circulating Nucleic Acid试剂盒提取血浆中ctDNA，定量仪检测ctDNA浓度，采用Wilcoxon非参数秩和检验进行相关比较，同时比较同时段采集、检测的癌胚抗原（CEA）手术前后浓度表达情况；② 采用Illumina HiSeq2000测序平台对癌组织、外周血细胞及ctDNA进行基因测序，并检测筛选出突变基因，以癌组织为阳性对照、术前血细胞为阴性对照，对ctDNA进行508个肿瘤常见相关基因突变情况进行了分析。结果：5例患者手术前以及手术后21-37天ctDNA浓度中位数分别为0.612、0.430ng/μL，二者差异有统计学意义（P<0.05），而同阶段手术前后CEA浓度改变未有统计学意义；在1例患者肺癌组织与术前ctDNA检测到多个相同突变，且术后ctDNA、血细胞样本均未见与术前样本有相同基因的同类型突变。结论：ctDNA与非小细胞肺癌组织突变类型具有一致性；ctDNA定量有望作为评估非小细胞肺癌肿瘤负荷变化的新分子生物学标志物。 |
| 英文摘要: |
| Objective The aim of this study was to study the quantitative expression of circulating tumour DNA
(ctDNA) in patients with non-small cell lung cancer (NSCLC) before and after radical operation and to
explore the correlation between gene mutations in non-small cell lung cancer tissues and those in ctDNA.
Methods We randomly assigned 5 NSCLC patients from the Department of Thoracic Surgery of Fujian
Medical University Union Hospital. All the patients had undergone radical surgery. Venous blood samples
were collected from the 5 NSCLC patients at two time points (before the operation and 21–37 days after the
operation) for monitoring ctDNA levels. This was done by isolating plasma from venous blood using high
velocity centrifugation, extracting DNA from the plasma using the QIAamp Circulating Nucleic Acid kit, and
then quantifying the ctDNA levels. The results were analyzed using the Wilcoxon Rank Sum Test. Moreover,
the ctDNA levels were compared with those of carcinoembryonic antigen (CEA), which was detected
simultaneously with the ctDNA. Then, DNA samples from the tumor tissues and peripheral blood cells and
ctDNA were sequenced using the Hiseq2000 sequencing platform (Illumina) and the mutant genes were
screened out. Mutations that occurred within the tumor tissues were used as positive control, whereas
those found in the pre-operative blood cells were used as a negative control. Based on the mutational
analysis of ctDNA genes, a total of 508 cancer-related genes were screened.
Results The median values of the pre- and post-operative ctDNA levels in the 5 patients with NSCLC
were 0.612 (0.518–0.876) and 0.430 (0.372–0.612) ng/μL, respectively. There was a significant difference
between the two groups (P < 0.05). The pre-operative CEA level was slightly higher than the post-operative
level (P > 0.05). In one of the cases, LC tissues showed multiple mutations, consistent with pre-operative
ctDNA. Moreover, isogenic mutations of the same type were not detected in post-operative ctDNA or
peripheral blood cells.
Conclusion Mutations found in the lung cancer (LC) ctDNA gene were consistent with the mutation
type of LC tissue. Hence, the quantitative and qualitative analysis of ctDNA is a promising novel molecular
biomarker for the evaluation of tumor burden changes in NSCLC. |
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