神经母细胞瘤组织及裸鼠荷瘤细胞RECK和MMP-14的表达及临床意义

目的检测RECK及膜型基质金属蛋白酶-14（MMP-14）在神经母细胞瘤患者蜡块标本中的表达，进一步检测将患者新鲜肿瘤标本原代培养建立的细胞系接种于裸鼠后自建的裸鼠原位荷瘤及转移瘤细胞系中二者的表达，比较裸鼠两种细胞中二者的表达差异及临床意义。方法首先采用PV-6000免疫组化方法检测临床及病理资料齐全的存档神经母细胞瘤蜡块标本36例及节细胞神经瘤蜡块标本10例中RECK及MMP-14蛋向的表达。其次应用RT-PCR法检测自建裸鼠原位荷瘤及转移瘤细胞中RECK及MMP-14mRNA的表达。结果RECK蛋白在神经母细胞瘤组织中低表达（16．7％），并随肿瘤浸润深度的加深及远处转移的发生而降低（P=0．015）。MMP-14蛋白在神经母细胞瘤组织中高表达（58．3％），并随肿瘤浸润深度的加深及远处转移的发生而增高（P=0．002）。裸鼠原位荷瘤细胞系中RECKmRNA的表达较裸鼠转移瘤细胞系中明碾增高（t=3．012，P〈0．05），MMP-14mRNA表达则明显降低（t=2．802，P〈0．05）。结论本研究提示，RECK及MMP-14蛋白和基因可能与神经母细胞瘤的浸润、转移有关，RECK及MMP-14可能在小儿神经母细胞瘤的发生、发展中起重要作用。

Expression and Significance of the Matrix Metalloproteinase Inhibitor RECK and MMP-14 in Neuroblastoma

Objective To investigate the expression of RECK and MMP-14 protein in human neuroblastoma, then establish the bear-tumor nude mice model of transplanted tumor in situ and metastatic tumor to detect the expression of RECK and MMP-14 mRNA, so we can discuss their roles in metastasis and development of the tumor. Methods PV-6000 immunohistochemistry was used to measure the expression of RECK and MMP-14 in 36 samples of neuroblastoma tissue selected from paraffin wax-embodied specimens with complete ,linicopathological data, and another 10 cases of ganglioneuroma tissue. The expression of RECK and MMP-14 mRNA in nude mice cell lines was measured by RT-PCR. Results The expression rate of RECK in neuroblastoma was low（16.7%）, and reduced with the increased invasive depth and long-distance metastasis（P=0.015 〈0.05）. The expression rate of MMP-14 in neuroblastoma was high （58.3%）, and elevated with the increased invasive depth and long-distance metastasis （P =0.002〈0.05）.The mRNA level of the RECK gene in the mouse primary tumor cell line was lower than that in the mouse metastatic tumor cell line. The mRNA level of the MMP-14 in the mouse primary tumor cell line was higher than that in the Mouse metastatic tumor cell line. Conclusions These results indicate that RECK and MMP-14 might play important role in the pathogenesis of neuroblastoma. RECK and MMP-14 could be considered as the marker for predicting the neuroblastoma metastasis and development.