RECK及MMP-2、MMP-9在神经母细胞瘤组织中的表达与肿瘤侵袭转移研究

Expressions of RECK, MMP-2 and MMP-9 and invasion of tumor metastasis in neuroblastoma

目的检测RECK基因、基质金属蛋白酶MMP-2及MMP-9在神经母细胞瘤组织中的表达,进一步探讨神经母细胞瘤发生、发展、侵袭和转移机制。方法(1)体内水平检测RECK、MMP-2、MMP-9基因在瘤组织中的表达:采用第二代通用型二步法监测系统(PV-6000)免疫组织化学方法对35例神经母细胞瘤患儿瘤体蜡块组织(转移组及无转移组)中的基因表达情况进行检测;(2)体外水平检测RECK、MMP-2、MMP-9基因在瘤组织中的表达:建立荷瘤裸鼠模型,获取新鲜肿瘤组织标本,采用SuperArray系列Oligo Tumor Metastasis Microarray基因芯片对标本进行芯片杂交,检测发光信号,通过图像及数据分析检测基因表达差异。结果(1)体内水平:人神经母细胞瘤组织RECK基因在转移组中的阳性表达率为6.7%,明显低于在无转移组中的表达,两组间差异有统计学意义(P=0.018)。MMP-2及MMP-9基因在转移组中的表达分别为86.7%及73.3%,明显高于在无转移组中的表达,两组间差异有统计学意义(P<0.05)。(2)体外水平:在荷瘤裸鼠神经母细胞瘤的肿瘤组织中,与原位瘤组织相比,转移瘤上调>2倍的基因为MMP-2、MMP-9,下调>2倍的基因为RECK。结论RECK、MMP-2、MMP-9基因有可能作为判断神经母细胞瘤的侵袭性和预后情况的参考指标,可能会作为其预测指标和干预靶点,对其诊断及治疗提供重要的指导作用。

Objective To explore the expressions of RECK, MMP-2 and MMP-9, and to examine the occurrence, development, invasion and metastatic mechanism of neuroblastoma. MethodsThe in vivo expressions of RECK, MMP-2 and MMP-9 were detected in tumor tissues.And PV-6000 immunohistochemistry was employed for detecting the expression levels in 35 samples of local in situ tumor and metastatic neuroblastoma tissues.The in vitro expressions of RECK, MMP-2 and MMP-9 were detected in tumor tissues.The model of nude mice was established.Then fresh specimens of neuroblastoma tissues were collected and hybridized by Oligo Tumor Metastasis Microarray of SuperArray.The luminescent signals of neuroblastoma tissues were detected and differentially expressed genes analyzed by images and data. ResultsThe positive expression rate of RECK protein was 6.7% in metastatic neuroblastoma tissues and it significantly decreased as compared with in situ neuroblastoma tissues.The inter-group differences were statistically significant (P=0.018). The positive expression rates of MMP-2 and MMP-9 proteins were 86.7% and 73.3% respectively and both significantly rose in metastatic neuroblastoma tissues.The inter-group differences were statistically significant (P<0.05). In neuroblastoma tissues of nude mice, as compared with in situ neuroblastoma tissues, genes up-regulated by >2 folds in metastatic neuroblastoma tissues included MMP-2, MMP-3, MMP-9, and gene down-regulated by >2 folds included RECK. ConclusionsAs the reference indices of invasion and prognosis of neuroblastoma, RECK, MMP-2 and MMP-9 may serve as predictors of neuroblastoma and intervention targets.This study provides important guidance in the diagnosis and treatment of neuroblastoma.