癌变·畸变·突变 ›› 2015, Vol. 27 ›› Issue (5): 332-340,346.doi: 10.3969/j.issn.1004-616x.2015.05.002

• 论著 • 上一篇    下一篇

结直肠癌淋巴结转移相关基因标志物的筛选

姚雨江1,2, 谢妮1, 刘丽桃1, 李仲航1, 万丽丽1, 朱婷1   

  1. 1. 深圳大学第一附属医院/深圳市第二人民医院, 广东 深圳 518035;
    2. 襄阳市中心医院, 湖北 襄阳 441021
  • 收稿日期:2015-05-20 修回日期:2015-07-29 出版日期:2015-09-30 发布日期:2015-09-30
  • 通讯作者: 谢妮,E-mail:kejiaoke100@163.com E-mail:kejiaoke100@163.com
  • 作者简介:姚雨江,Tel:18327528102;E-mail:xinxinyyj@126.com。
  • 基金资助:

    广东省深圳市技术研究计划基础研究项目(JCYJ20120613171430264);广东省科技计划项目(2014A020212038,2013B021800097)

Screening of genetic markers associated with lymph node metastasis in colorectal cancer

YAO Yujiang1,2, XIE Ni1, LIU Litao1, LI Zhonghang1, WAN Lili1, ZHU Ting1   

  1. 1. The First Affiliated Hospital of Shenzhen University/Shenzhen Second People's Hospital, Shenzhen 518035, Guangdong;
    2. Xiangyang Central Hospital, Xiangyang 441021, Hubei, China
  • Received:2015-05-20 Revised:2015-07-29 Online:2015-09-30 Published:2015-09-30

摘要:

目的:筛选与结直肠癌淋巴结转移相关的基因或肿瘤标志物的单核苷酸多态性(SNPs)、拷贝数变异(CNVs)并检测其mRNA表达量。方法:从前期收集的1 053个肿瘤相关候选基因中,采用目标区域测序(Illumina Hiseq)捕获25例结直肠病人的癌组织和癌旁组织中与淋巴结转移相关的候选基因的SNPs和CNVs;用荧光定量PCR检测39例结直肠癌病人的癌组织和癌旁组织中6个相关候选基因(VEGFCCCNA2IL2ABCG2EGFNFKB1)mRNA表达量的改变。结果:SLC28A3BRCA1RRM2PMS2CDAEPHX1RALYCD33BCL10ETV1MST1RKMT2BBCL2LSM3TTF1MAP3K1基因的SNPs与结直肠癌的淋巴结转移相关,差异具有统计学意义(P均< 0.05);DDR1CYP21A2SULT1A1XRCC2POU5F1FLT1基因的CNVs在淋巴结转移组和未转移组均未见明显差异(P均>0.05); 与无淋巴结转移组相比,EGF和NFKB1基因的mRNA表达量在淋巴结转移组中降低,差异具有统计学意义(P<0.05)。结论:SLC28A3BRCA1RRM2PMS2CDAEPHX1RALYCD33BCL10ETV1MST1RKMT2BBCL2LSM3TTF1MAP3K1基因的SNPs以及EGFNFKB1基因表达下调可能与结直肠癌淋巴结转移有关,为结直肠癌淋巴结转移的潜在分子标志。本实验未发现候选基因CNVs与结直肠癌病人的淋巴结转移相关。

关键词: 单核苷酸多态性, 拷贝数变异, mRNA表达量, 结直肠癌, 淋巴结转移

Abstract:

OBJECTIVE: Try to identify single nucleotide polymorphism (SNPs), copy number variation (CNVs) and mRNA expression from the genetic or tumor markers that are associated with lymph node metastases in colorectal cancer. METHODS:Targeted next-generation sequencing(Illumina Hiseq) was applied to capture SNPs and CNVs in tumor-related candidate genes of tumor tissues and paired normal tissues adjacent to the tumors from 25 colorectal cancer specimens;real time-PCR was used to detect specific mRNA expression of tumor-related candidate genes (VEGFCCCNA2IL2ABCG2EGF and NFKB1) on chromosome 4 of 39 colorectal cancer patient specimens. RESULTS: The SNPs in SLC28A3, BRCA1, RRM2, PMS2, CDA, EPHX1, RALY, CD33, BCL10, ETV1, MST1R, KMT2B, BCL2, LSM3, TTF1, MAP3K1 genes were significantly correlated with lymphatic metastasis risk (P<0.05). There were no significant differences of the CNVs in the DDR1, CYP21A2, SULT1A1, XRCC2, POU5F1, FLT1 genes between the positive lymph node metastasis group and the negative lymph node metastasis group(P>0.05). Compared with non lymph node metastasis group, mRNA expressions of EGF and NFKB1 were both down-regulated in the lymph node metastasis group, and the difference was statistically significant(P<0.05). CONCLUSION:SNPs in SLC28A3, BRCA1, RRM2, PMS2, CDA, EPHX1, RALY, CD33, BCL10, ETV1, MST1R, KMT2B, BCL2, LSM3, TTF1, MAP3K1 genes and down-regulated expression of EGF and NFKB1 might be the potential genetic markers in lymph node metastasis of colorectal cancers. No CNV was found to be associated with lymph node metastasis in colorectal cancer in our study.

Key words: single nucleotide polymorphism, copy number variation, mRNA expression, colorectal cancer, lymph node metastasis

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