Sehgal A, Boynton A L, Young R F, Vermeulen S S, Yonemura K S, Kohler E P, Aldape H C, Simrell C R, Murphy G P
Deke Slayton Center for Brain Cancer Studies, Pacific Northwest Cancer Foundation, Northwest Hospital, Seattle, WA 98125, USA.
Int J Cancer. 1998 May 18;76(4):451-8. doi: 10.1002/(sici)1097-0215(19980518)76:4<451::aid-ijc1>3.0.co;2-q.
Using the technique of differential display-polymerase chain reaction (DD-PCR), we isolated a cDNA fragment that is over-expressed in glioblastoma multiforme tissue as compared to normal brain tissue. Sequence analysis indicated that this sequence is identical to the previously isolated human neuron-glia-related cell adhesion molecule hNr-CAM. Gene-specific RT-PCR analysis indicated that hNr-CAM is over-expressed in high-grade astrocytomas, gliomas and glioblastoma tumor tissues as compared to normal brain tissue. High levels of hNr-CAM expression also were observed in cell lines derived from astrocytomas, gliomas and glioblastoma multiforme tumors. Low levels of hNr-CAM expression were observed in neuroblastoma, meningiomas, melanoma, normal breast and prostate tumor tissues. Northern blot analysis showed an alternatively spliced mRNA of 1.4 kb in several tumors as compared to the 7.5 kb transcript found in normal brain tissue. Genomic Southern blot analysis of DNA from 3 brain tumor cell lines showed that over-expression of hNr-CAM in brain tumors was not due to gene amplification. In situ hybridization analysis indicated that 11 of the 20 human brain tumor samples studied showed hNr-CAM over-expression. Our results suggest that hNr-CAM is over-expressed in malignant brain tumors and can serve as a novel marker for brain tumor detection and perhaps therapy.
运用差异显示聚合酶链反应(DD-PCR)技术,我们分离出了一个在多形性胶质母细胞瘤组织中相对于正常脑组织过度表达的cDNA片段。序列分析表明,该序列与先前分离出的人类神经元-神经胶质相关细胞粘附分子hNr-CAM相同。基因特异性逆转录-聚合酶链反应(RT-PCR)分析表明,与正常脑组织相比,hNr-CAM在高级别星形细胞瘤、神经胶质瘤和胶质母细胞瘤肿瘤组织中过度表达。在源自星形细胞瘤、神经胶质瘤和多形性胶质母细胞瘤肿瘤的细胞系中也观察到高水平的hNr-CAM表达。在神经母细胞瘤、脑膜瘤、黑色素瘤、正常乳腺和前列腺肿瘤组织中观察到低水平的hNr-CAM表达。Northern印迹分析显示,与在正常脑组织中发现的7.5 kb转录本相比,几种肿瘤中有一个1.4 kb的可变剪接mRNA。对3个脑肿瘤细胞系的DNA进行基因组Southern印迹分析表明,脑肿瘤中hNr-CAM的过度表达不是由于基因扩增。原位杂交分析表明,在所研究的20个人类脑肿瘤样本中,有11个显示hNr-CAM过度表达。我们的结果表明,hNr-CAM在恶性脑肿瘤中过度表达,可作为脑肿瘤检测乃至治疗的一种新标志物。