[1]靳 杨,姜利琼,房 烨,等.结直肠癌患者血清外泌体与组织中KRAS，BRAF，NRAS 和PIK3CA 基因突变检测及临床意义的比较[J].现代检验医学杂志,2023,38(01):22-26.[doi:10.3969/j.issn.1671-7414.2023.01.005]
　JIN Yang,JIANG Li-qiong,FANG Ye,et al.Comparison of KRAS, BRAF, NRAS and PIK3CA Gene Mutations in Serum Exosomes and Tissues of Patients with Colorectal Cancer and Their Clinical Significance[J].Journal of Modern Laboratory Medicine,2023,38(01):22-26.[doi:10.3969/j.issn.1671-7414.2023.01.005]
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结直肠癌患者血清外泌体与组织中KRAS，BRAF，NRAS 和PIK3CA 基因突变检测及临床意义的比较()
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《现代检验医学杂志》[ISSN:/CN:]

卷:
第38卷
期数:
2023年01期
页码:
22-26
栏目:
论著
出版日期:
2023-01-15

文章信息/Info

Title:
Comparison of KRAS, BRAF, NRAS and PIK3CA Gene Mutations in Serum Exosomes and Tissues of Patients with Colorectal Cancer and Their Clinical Significance
文章编号:
1671-7414（2023）01-022-05
作者:
靳 杨1; 姜利琼2; 房 烨1; 曾燕兰1; 何 毅1; 边革元1
(1. 中国人民解放军联勤保障部队第九二〇医院重症医学科,昆明650032；2. 云南省阜外心血管病医院健康管理中心，昆明650102 )
Author(s):
JIN Yang1; JIANG Li-qiong2; FANG Ye1; ZENG Yan-lan1; HE Yi1; BIAN Ge-yuan1
（1. Department of Critical Care Medicine, No. 920 Hospital, Joint Logistics Support Force of the Chinese People’s Liberation Army, Kunming 650032, China；2 .Health Management Center, Fuwai Cardiovascular Hospital of Yunnan Province, Kunming 650102, China）
关键词:
结直肠癌; 血清外泌体; 基因突变
分类号:
R735.3；R730.43
DOI:
10.3969/j.issn.1671-7414.2023.01.005
文献标志码:
A
摘要:
目的 检测结直肠癌(colorectal cancer, CRC) 患者血清外泌体KRAS，BRAF，NRAS 和PIK3CA 相关基因位点的突变，分析其与癌组织基因突变的一致性及其可能的影响因素。方法 2019 年2 月～ 2021 年1 月中国人民解放军联勤保障部队第九二〇医院100 例结直肠癌患者作为研究对象。提取患者血清外泌体，并用蛋白免疫印迹（Westernblot，WB）法检测外泌体标志物CD63 和TSG101 的蛋白表达；聚合酶链式反应（polymerase chain reaction，PCR）检测血清外泌体及手术切除的癌组织中KRAS，BRAF，NRAS 和PIK3CA 基因突变情况，Kappa 一致性检验分析血清外泌体突变与组织突变的一致性，Logistic 单因素回归分析影响一致性的因素。结果 癌症基因组图谱（the cancer genomeatlas，TCGA）数据显示，KRAS，BRAF，NRAS 和PIK3CA 的结直肠癌突变率分别为35% ～ 96%，5% ～ 15%，5% ～ 30%和18% ～ 36%，且KRAS 和BRAF 的突变与结直肠癌患者的低存活率有关。血清外泌体中KRAS，BRAF，NRAS 和PIK3CA 的突变率分别为94.00%，11.00%，17.00% 和35.00%；结直肠癌组织KRAS，BRAF，NRAS 和PIK3CA 的突变率分别为34.00%，5.00%，6.00% 和16.00%；血清外泌体中KRAS，BRAF，NRAS 和PIK3CA 的突变率均高于组织，差异有统计学意义（χ2=101.027 ～ 256.250，均P ＜ 0.05）。血清外泌体与组织中KRAS 突变的一致率为40.00%（Kappa值=0.064，P>0.05），BRAF 突变的一致率为99.00%（Kappa 值=0.599，P<0.05），NRAS 突变的一致率为89.00%（Kappa 值=0.475，P<0.05），KRAS 突变的一致率为81.00%（Kappa 值=0.523，P<0.05）。ECOG 评分、转移、临床分期是影响外泌体和组织KRAS，BRAF，NRAS 和PIK3CA 的突变检测一致性的因素。结论 结直肠癌患者血清外泌体KRAS，BRAF，NRAS 和PIK3CA 的突变率高于组织，二者BRAF，NRAS 和PIK3CA 的一致性中等，为外泌体的基因检测指导临床靶向治疗提供参考。
Abstract:
Objective To detect the mutations loci in exosomes related gene of KRAS, BRAF, NRAS and PIK3CA in patients with colorectal cancer(CRC), and analyze the consistency with cancer tissue gene mutations and its possible influencing factors. Methods From February 2019 to January 2021, 100 patients with colorectal cancer in the No. 920 Hospital of the Joint Logistics Support Force of the Chinese People’s Liberation Army were selected as the research object. The serum exosomes of patients were extracted, and the protein expressions of exosomes markers CD63 and TSG101 were detected by Western blot (WB).Detection of KRAS, BRAF, NRAS and PIK3CA gene mutations in serum exosomes and surgically resected cancer tissues by polymerase chain reaction (PCR). Kappa consistency test was used to analyze the consistency between serum exosome mutations and tissue mutations. Logistic univariate regression was used to analyze the factors affecting the consistency. Results The cancer genome atlas (TCGA) data showed that the mutation rates of KRAS, BRAF, NRAS and PIK3CA were 35% ～ 96%， 5% ～ 15%，5% ～ 30% and 18% ～ 36%， respectively, and the mutations of KRAS and BRAF were related to the low survival rate of colorectal cancer patients. The mutation rates of KRAS, BRAF, NRAS and PIK3CA in serum exosomes were 94.00%, 11.00%, 17.00% and 35.00% respectively. The mutation rates of KRAS, BRAF, NRAS and PIK3CA in tissues were 34.00%, 5.00%, 6.00% and 16.00% respectively. The results of tissue gene testing were provided by the clinical pathology laboratory，the mutation rates of KRAS, BRAF, NRAS and PIK3CA in serum exosomes were higher than those in tissues, and the differences were statistically significant(χ2=101.027 ～ 256.250，all P < 0.05). The consistency rate of KRAS mutation in serum exosomes and tissues was 40.00% (Kappa value =0.064, P>0.05), BRAF mutation was 99.00% (Kappa value =0.599, P<0.05), NRAS mutation was 89.00% (Kappa value =0.475, P<0.05), and KRAS mutation was 81.00% (Kappa value =0.523, P<0.05). The consistency of ECOG score, metastasis and staging of patients is high. Conclusion The mutation rates of serum exosomes KRAS, BRAF, NRAS and PIK3CA in colorectal cancer patients were higher than those in tissues, and the consistency of BRAF, NRAS and PIK3CA was moderate, which may provide a reference for exosomes gene detection to guide clinical targeted therapy.

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备注/Memo

备注/Memo:
基金项目： 联勤保障部队第九二〇医院院内科技计划《外泌体DNA 检测指导大肠癌患者个体化治疗的可行性研究》，课题编号：2019YGC01。
作者简介：靳杨（1987-），女，博士，主治医师，研究方向：肿瘤精准医疗及危重症救治，E-mail:fecesathe70@21cn.com。
通讯作者：边革元（1966-），男，本科，主任医师，研究方向：肿瘤危重症救治，E-mail:fecesathe70@21cn.com。
更新日期/Last Update: 2023-01-15