大肠癌早期肝转移的血清特征蛋白及组织表达特异基因筛查

Screening of serum proteomic patterns and tissue-specific genes for early liver metastasis of colorectal cancer

目的应用表面增强激光解吸电离飞行时间质谱(SELDI-TOF-MS)技术及RT2 ProfilerTM PCR Array HumanTumor Metastasis(PAHS-028A)芯片筛查早期肝转移患者血清特异性蛋白及相关肿瘤基因,为大肠癌肝转移的早期诊断提供理论依据。方法应用弱阳离子蛋白质芯片(WCX2)及SELDI-TOF-MS技术检测20例大肠癌和20例大肠癌早期肝转移患者血清中蛋白的相对含量。PCR基因芯片技术对原发灶和肝转移灶组织进行差异基因筛选。结果应用SELDI-TOF-MS的3组样本质荷比在2 000～30 000间有3 774、11 851的2个蛋白峰有显著差异。应用RT2 ProfilerTM PCR Array HumanTumor Metastasis(PAHS-028A)芯片技术发现在大肠癌原发灶中表达明显高于肝转移结节的基因有:ACTB、APC、CTNNA1、ITGB3、NR4A3、MMP10、MMP11、ETV4、CTSL1、RB1、GNRH1、HPSE、CDKN2A、KISS1R、IL8RB、ITGA7、DENR、RPSA、CX-CR4、NME2、PNN、SMAD4、SRC、SSTR2、SYK、RORB、TCF20、MMP3、MYCL1、TIMP2、TIMP3、TIMP4、TRPM1;在转移灶表达明显高于原发灶的基因有:MMP9、FN1、CST7、CCL7。结论利用SELDI-TOF-MS及基因芯片技术可以为大肠癌早期肝转移的机制研究及诊断与治疗提供实验数据和资料。

Objective To screen for the serum proteomic patterns and related genes in early liver metastasis of colorectal cancer by surface enhanced laser desorption/ionization time of flight mass spectrometry（SELDI-TOF-MS） and the RT2 ProfilerTM PCR Array Human Tumor Metastasis（PAHS-028A） chip,so as to provide theoretical evidence for diagnosis of early liver metastasis of colorectal cancer.Methods The serum proteins of 20 colorectal cancer patients and 20 patients with early liver metastasis of colorectal cancer were detected by WCX2 chip and SELDI-TOF-MS.PCR gene chips were used to screen the differentially expressed genes between the primary tumor and the liver metastases.Results SELDI-TOF-MS found that,when the M/Z values ranged 2 000-30 000,the contents of two proteins（3 774 and 11 851） were significant different in three samples.PCR gene chip found that the expressions of following genes were significantly higher in the primary colorectal cancer specimens than in the liver metastatic nodules： ACTB,APC,CTNNA1,NR4A3,MMP10,CTSL1,RB1,HPSE,ETV4,GNRH1,CDKN2A,KISS1R,IL8RB,ITGA7,ITGB3,DENR,RPSA,CXCR4,MYCL1,NME2,PNN,SMAD4,MMP11,SRC,RORB,SSTR2,SYK,TCF20,MMP3,TIMP2,TIMP3,TIMP4,and TRPM1;and the following genes were significantly higher in the liver metastases than in the primary tumors： MMP9,FN1,CST7,and CCL7.Conclusion SELDI-TOF-MS and gene chip technique can provide a theoretic basis for the mechanism,diagnosis and treatment of early liver metastasis of colorectal cancer.