结直肠癌淋巴结转移相关基因标志物的筛选  

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

作　　者：姚雨江 谢妮[1] 刘丽桃 李仲航 万丽丽[1] 朱婷[1] 

机构地区：[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

出　　处：《癌变．畸变．突变》2015年第5期332-340,346,共10页Carcinogenesis,Teratogenesis & Mutagenesis

基　　金：广东省深圳市技术研究计划基础研究项目(JCYJ20120613171430 264);广东省科技计划项目(2014A020212038;2013B021800097)

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m RNA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(VEGFC、CCNA2、IL2、ABCG2、EGF及NFKB1)m RNA表达量的改变。结果:SLC28A3、BRCA1、RRM2、PMS2、CDA、EPHX1、RALY、CD33、BCL10、ETV1、MST1R、KMT2B、BCL2、LSM3、TTF1、MAP3K1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);DDR1、CYP21A2、SULT1A1、XRCC2、POU5F1、FLT1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m RNA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:SLC28A3、BRCA1、RRM2、PMS2、CDA、EPHX1、RALY、CD33、BCL10、ETV1、MST1R、KMT2B、BCL2、LSM3、TTF1、MAP3K1基因的SNPs以及EGF、NFKB1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。OBJECTIVE： Try to identify single nucleotide polymorphism（SNPs）,copy number variation（CNVs） and m RNA 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 m RNA expression of tumor-related candidate genes（VEGFC, CCNA2,IL2,ABCG2,EGF 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, MST1 R, KMT2 B,BCL2,LSM3,TTF1,MAP3K1 genes were significantly correlated with lymphatic metastasis risk（P0.05）. 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（P0.05）. Compared with non lymph node metastasis group,m RNA expressions of EGF and NFKB1 were both down-regulated in the lymph node metastasis group,and the difference was statistically significant（P0.05）. CONCLUSION：SNPs in SLC28A3, BRCA1,RRM2,PMS2,CDA,EPHX1,RALY,CD33, BCL10,ETV1,MST1 R,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.

关 键 词：单核苷酸多态性 拷贝数变异 m RNA表达量 结直肠癌 淋巴结转移 

分 类 号：R735.3[医药卫生—肿瘤]