Migratory, invasive and metastatic capacity of NCAM transfected rat glioma cells

A cDNA encoding a transmembrane 140 kDa isoform of the neural cell adhesion molecule, NCAM, was transfected into the rat glioma cell line BT4Cn. Transfectants with a homogeneously high expression of NCAM-B showed a decreased capacity for penetration of an artificial basement membrane when compared to cells transfected with expression-vector alone or untransfected cells. However, when injected subcutaneously into nude mice, both NCAM expressing cells and control cells produced invasive tumors. Nude mice injected with NCAM positive cells developed tumors with slower growth rates as compared to those induced by NCAM negative cells. This implies that NCAM may not only be involved in adhesive and motile behaviour of glioma cells, but also in their growth regulation.     

Overexpression of the 18A2/mts1 gene and down–regulation of the TIMP–2 gene in invasive human glioma cell lines in vitro

Invasion of the reconstituted extracellular matrix composite, Matrigel, by eight human glioma–derived cell lines and human fetal brain cells was assessed in vitro using 8 um polycarbonate filters in a modified Boyden migration chamber. With the exception of one low grade glioma derived cell line, all lines studied proved to be invasive while normal fetal brain cells failed to invade. This invasive potential was independent of the histological grade of the tumour from which the cell lines originated. In addition, the expression of the metastasis–associated gene 18A2lmts1 as well as the tissue inhibitor of metalloproteinases–2 (TIMP–2) was analysed in each of the glioma–derived cell lines. The 18A2/mtsl was expressed in all the cells studied with the exception of fetal brain cells and the low grade non–invasive glioma derived IPRK–7 cell line. The 18A2/mtsl related genes coding for the S100 subfamily of calcium binding proteins were found to be differentially and overexpressed in invasive cell lines. TIMP–2 was expressed only in noninvasive cell lines. These results suggest that the 18A2/ mtsl and TIMP–2 genes could play an important role in the invasive behaviour of human glioma cells in vitro. .     

Inward rectifying potassium channels in human malignant glioma cells

Human glioma cells obtained from established cell lines (Tp-276MG, Tp-301MG, Tp-378MG, Tp-483MG and U-251MG) were analyzed for the presence of ion channels with the tight-seal voltage clamp technique. The current-voltage relation revealed a marked inward rectification at hyperpolarizing voltages, due to the presence of inward rectifying K-channels in cells from all studied cell lines. These channels were conducting when the membrane potential was more negative than the K-equilibrium potential. The slope conductance for the inward K-currents (g_Ki) was affected both by [K⁺]_i and [K⁺]₀. g_Ki was proportional to [K⁺]₀ raised to 0.35 or 0.50, of which the larger value was measured in the presence of low [K⁺]_i (25mM). The rectification was not significantly different in cells perfused with Mg-free EDTA-buffered internal solution. Tl⁺ was 3.5 times more permaant than K⁺. g_ki was blocked by Cs⁺ (1 mM) in a voltage-dependent way (more effective in the hyperpolarized membrane), and by Na⁺ (154 mM) depending on voltage and time. From measurements of unitary current events in membrane patches (outside out or cell attached) the conductance of the single inward rectifying channel was estimated to be 27 ± 7 pS. This type of ion channel may be important for K-uptake by glial cells and hence for the K-homeostasis in the brain.     

Long-term follow-up results of 175 patients with malignant glioma: Importance of radical tumour resection and postoperative adjuvant therapy with interferon,ACNU and radiation

Summary We analysed long-term follow-up results of 175 patients with malignant glioma (110 glioblastoma and 65 anaplastic astrocytoma) treated under five different regimes during the past two decades. The factors of age (less than 40), histology (anaplastic astrocytoma) and type of adjuvant therapy (radiation and chemotherapy) contributed to long survival. The other important factor was the response to adjuvant therapy.Cases of gross total removal or complete response (CR) of a residual tumour to an adjuvant therapy showed a better prognosis. The three and five year survival rate was 42% and 24%, respectively. The highest CR ratio (23%) was seen in patients treated by intravenous injection of interferon and ACNU in addition to radiotherapy (IAR therapy).     

Primary diffuse leptomeningeal gliomatosis followed with serial magnetic resonance images

We report a case of primary diffuse leptomeningeal gliomatosis (PDLG) followed up with serial magnetic resonance images (MRI). A 45‐year‐old man manifested with bilateral abducens nerve palsy and meningisms. Repeated MRI revealed diffuse leptomeningeal enhancement throughout the central nervous system without intra‐axial mass accompanied with the dilatation of ventricles and focally enlarged cerebral sulci. Brain biopsies showed a leptomeningeal gliomatosis. The MRI findings described here would contribute to the diagnosis of PDLG among other common diseases diffusely spreading along the leptomeningeal structures.     

术中磁共振影像神经导航治疗脑胶质瘤的临床初步应用(附61例分析)

目的总结术中磁共振影像(iMRI)神经导航手术治疗脑胶质瘤的初步经验。方法2006年3月～2006年12月,61例脑胶质瘤病人接受iMRI神经导航手术。结果手术总耗时2.5～8.5 h,平均(5.2±1.5)h。图像质量良好49例(80.3%),一般7例(11.5%),差5例(8.2%)。56例图像质量优良的病人中,iMRI扫描次数2～5次,平均(2.6±0.8)次;其中2次32例,3次16例,4次5例,5次3例;24例(42.9%)经iMRI发现肿瘤切除范围未达术前计划,仍需进一步切除。无iMRI相关不良事件发生。结论iMRI神经导航手术治疗脑胶质瘤安全、有效,可实时纠正术中脑移位误差,精确定位脑胶质瘤的影像学边界,定量评估手术切除范围,有效提高肿瘤切除率。     

C－myc反义寡聚脱氧核苷酸抑制人脑胶质瘤细胞系BT_（325）Myc蛋白合成和细胞增殖并诱导其分化的研究

目的：探讨在体外ｃ－ｍｙｃ反义核酸是否可通过阻断人脑胶质瘤细胞中ｃ－ｍｙｃ基因的表达而抑制细胞增殖并诱导分化．方法：人工合成与ｃ－ｍｙｃｍＲＮＡ起始码及其后四个密码子互补的寡聚脱氧核苷酸（简称反义核酸）片段，用它处理培养的ＢＴ３２５细胞，观察它对细胞增殖的影响．同时用免疫细胞化学方法检测细胞中Ｍｙｃ蛋白的水平以及能反映胶质瘤细胞分化的Ｓ－１００和ＧＦＡＰ两种蛋白的水平，分析这些指标的变化．结果：发现４ｕｍｏｌ／Ｌ的ｃ－ｍｙｃ反义核酸明显抑制ＢＴ３２５细胞的增殖和Ｍｙｃ蛋白的合成，且后者发生在加入反义核酸后１ｈ，并持续２４ｈ以上，而细胞增殖受抑制要到第５日才明显．从第２日到第５日细胞中Ｓ－１００和ＧＦＡＰ染色明显加深，反映细胞有分化趋势．用同样长度的无关序列寡聚脱氧核苷酸作对照，则未见上述变化，表明ｃ－ｍｙｃ反义核酸的作用是序列特异性的．结论：ｃ－ｍｙｃ反义核酸可特异地抑制ＢＴ３２５细胞中Ｍｙｃ蛋白的合成和细胞增殖，并能诱导其分化．     

人脑胶质瘤中P53和VEGF 的表达及意义

目的 探讨胶质瘤组织中血管内皮生长因子(VEGF)和P53蛋白的表达及其临床意义。方法 应用免疫组织化学技术对50例胶质瘤组织进行P53、VEGF表达的检测。结果 P53、VEGF的表达均随胶质瘤病理级别的升高而升高。Ⅰ～Ⅳ级病理分级中P53阳性率分别为22．22％(4／18)，65％(13／20)，75％(9／12)；VEGF阳性率分别为27．78％(5／18)，90％(18／20)，100％(12／12)。P53、VEGF的表达均与胶质瘤病理分级显著相关(P＜0．01)，P53和VEGF阳性表达符合率为76％(38／50)，两者的表达有显著性相关(P＜0．01)。结论 P53和VEGF蛋白的表达是判断胶质瘤生物学行为的重要指标；胶质瘤组织突变的P53基因可上调VEGF的表达，促进血管生成，进而影响胶质瘤的进展。     

人脑胶质瘤中cyclinD1/bcl-1和p27/kip1的表达与病理、预后的相关性研究

目的 :研究cyclinD1 bcl 1和p2 7 kip1在脑胶质瘤中表达及其与病理分级和患者预后的关系。方法 :用免疫法组化技术对 4 8例不同恶性程度 (WHO分类法 )的脑胶质瘤组织和 12例非肿瘤组织中cyclinD1 bc1 1和p2 7 kip1蛋白的表达作了检测 ,用图像分析系统定量分析 ,并与临床资料紧密相联系进行统计学处理。结果 :两种蛋白的免疫反应复合物定位于细胞核。其在脑胶质瘤中的阳性表达都高于非肿瘤组织 (P <0 0 5 ) ,cyclinD1 bcl 1阳性颜色的数量和强度都随肿瘤的恶性程度升高而增加 (P <0 0 5 ) ,相反p2 7 kip1阳性染色的数量和强度都随肿瘤的恶性程度升高而降低 (P <0 0 5 )。cyclinD1 bcl 1的高表达或 和p2 7 kip1的低表达预示着预后差。结论 :CyclinD1 bcl 1和p2 7 kip1的异常表达可能与脑胶质瘤的发生发展密切相关 ,二者可能有协同作用。二者的表达都可作为判断预后的一个指标