Ishii N, Tada M, Sakuma S, Sawamura Y, Shinohe Y, Abe H
Department of Neurosurgery, University of Hokkaido, School of Medicine, Sapporo, Japan.
J Neurooncol. 1998 Mar;37(1):17-23. doi: 10.1023/a:1005959719927.
We investigated expression of macrophage inflammatory protein-1 (MIP-1) alpha and beta in human astrocytoma cell lines and surgical specimens of astrocytic tumors. Enzyme-linked immunosorbent assay (ELISA) showed constitutive secretion of MIP-1alpha protein in only one and MIP-1beta in none of 7 cell lines tested. However, MIP-1alpha production was increased in three cell lines by stimulation with lipopolysaccharide (LPS) and 5 cell lines by stimulation with phorbol-12myristate-13-acetate (PMA). Also, induction of MIP-1beta production was observed in one cell line with LPS stimulation and in two cell lines with PMA stimulation. Reverse-transcription polymerase chain reaction (RT-PCR) showed the increase of MIP-1alpha and beta mRNA expression in these cell lines. The increase of the mRNA with the stimuli was further confirmed by Northern blot analysis. In contrast, RT-PCR analysis revealed that the majority of the tested tumor specimens of high-grade.astrocytomas expressed both MIP-1alpha and MIP-1beta mRNAs. ELISA detected MIP-1beta protein in 1 of 11 cerebrospinal fluid samples from patients with high-grade astrocytoma and in 8 of 9 tumor cyst fluid samples, whereas MIP-1alpha was detected in only 1 cyst fluid somple. Taken together, these results indicate that astrocytic tumor cells are capable of expressing and producing MIPs, and suggest that MIPs may participate in the inflammatory responses commonly seen in astrocytic tumors.
我们研究了巨噬细胞炎性蛋白-1(MIP-1)α和β在人星形细胞瘤细胞系及星形细胞肿瘤手术标本中的表达。酶联免疫吸附测定(ELISA)显示,在检测的7种细胞系中,只有1种组成性分泌MIP-1α蛋白,没有一种分泌MIP-1β蛋白。然而,用脂多糖(LPS)刺激后,3种细胞系中MIP-1α的产生增加;用佛波醇-12-肉豆蔻酸酯-13-乙酸酯(PMA)刺激后,5种细胞系中MIP-1α的产生增加。此外,在1种细胞系中观察到LPS刺激诱导MIP-1β产生,在2种细胞系中观察到PMA刺激诱导MIP-1β产生。逆转录聚合酶链反应(RT-PCR)显示这些细胞系中MIP-1α和β mRNA表达增加。Northern印迹分析进一步证实了刺激后mRNA的增加。相比之下,RT-PCR分析显示,大多数高级别星形细胞瘤的检测肿瘤标本同时表达MIP-1α和MIP-1β mRNA。ELISA在11份高级别星形细胞瘤患者的脑脊液样本中的1份以及9份肿瘤囊液样本中的8份中检测到MIP-1β蛋白,而仅在1份囊液样本中检测到MIP-1α。综上所述,这些结果表明星形细胞瘤细胞能够表达和产生MIPs,并提示MIPs可能参与星形细胞瘤中常见的炎症反应。