Evaluation of ACNU efficacy on patients with malignant glioma using sister chromatid exchange.

本项目的目的是利用姐妹染色单体互换(SCE)作为评估亚硝脲(ACNU)治疗恶性胶质瘤患者的疗效的一种方法。相关研究结果如下：1)培养的C6大鼠胶质瘤细胞的SCE随ACNU浓度的增加而增加，从7.31个/细胞(对照组)增加到35.2个/细胞(ACNU：3.0µg/ml)。SCES的增加与C6细胞集落形成实验结果有很好的相关性，并在较低浓度的ACNU下检测到细微的变化。还在继续研究建立从手术切除的胶质瘤组织中检测早期传代培养的SCES的方法。2)经植物血凝素激活的正常人外周血淋巴细胞经ACNU(3.0µg/ml)处理后，其SCE由未处理的7.6个/细胞增加到35.2个/细胞。ACNU处理后，细胞在无药物培养液中孵育后，SCES降低(损伤恢复)。3)对接受ACNU化疗的恶性脑胶质瘤患者的外周血淋巴细胞进行了SCES检测。静脉注射ACNU后1d至75d，SCES显著升高(从6.3~9.1个/细胞增加到17.0~46.0个/细胞)。在ACNU后的早期检查中发现了更多的SCE。鞘内注射ACNU后，SCES略有增加，为11.7~16.4个/细胞，而放射治疗后无明显变化。结果表明，检测外周血淋巴细胞SCES可检测ACNU治疗后对正常宿主细胞的损伤情况，可用于监测ACNU治疗恶性脑胶质瘤的临床疗效。正在进行一项研究，以评估我们是否可以通过估计计划接受ACNU治疗的患者的淋巴细胞的体外SCE来预测临床结果。

The aim of this project was to utilize the sister chromatid exchange (SCE) as a way of evaluating the efficacy of nitrosourea (ACNU) on patients with malignant glioma. Pertinent findings are described as below; 1) SCEs of C6 rat glioma cells in culture increased dependent on the ACNU concentration from 7.31/cells (control) to 35.2/cells (ACNU: 3.0mug/ml). the increase of SCEs correlated well with the results of the colony formation assay of C6 cells and detected subtle changes at the lower concentration of ACNU. Still going on the studies to establish the method measuring SCEs of early passage culture from glioma tissues obtained by surgical resection. 2) SCEs of normal peripheral lymphocytes activated by phytohemagglutinin increased from 7.6/cells (no treatment) to 35.2/cells by in vitro treatment with ACNU (3.0mug/ml). When cells were incubated in the drug-free medium after ACNU treatment, SCES decreased (the recovery from injury). 3) We measured SCEs of peripheral lymphocytes of patients with malignant glioma who underwent ACNU administration as postoperative chemotherapy. SCEs increased significantly from a day to 75 days after intravenous injection of ACNU (from 6.3-9.1/cells to 17.0-46.0/cells). A larger number of SCEs was detected when examined in the earlier period after ACNU. SCEs increased moderately to 11.7-16.4/cells after intrathecal administration of ACNU, but they did not after radiation therapy. We showed that measuring SCEs of peripheral lymphocytes detects the damage of normal host cells after the treatment with ACNU and would be useful in monitoring clinical efficacy of ACNU on patients with malignant glioma. In progress is the investigation to evaluate if we could predict the clinical results by estimating in vitro SCEs of lymphocytes from patients scheduled to have the treatment with ACNU.