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

本项目旨在利用姐妹染色单体交换（SCE）作为评估亚硝基脲（ACNU）治疗恶性胶质瘤疗效的一种方法。有关调查结果说明如下：1)培养C6大鼠胶质瘤细胞的SCEs随ACNU浓度的增加而增加，从7.31/细胞（对照组）增加到35.2/细胞（ACNU: 3.0mug/ml）。SCEs的增加与C6细胞集落形成实验结果有良好的相关性，并且在较低浓度的ACNU下检测到细微的变化。建立手术切除获得的胶质瘤组织早期传代培养物的SCEs测定方法仍在研究中。2)经植物血凝素激活的正常外周血淋巴细胞SCEs由未处理时的7.6/细胞增加到体外ACNU （3.0mug/ml）处理后的35.2/细胞。ACNU处理后的细胞在无药培养基中孵育，SCES降低（损伤恢复）。3)我们测量了恶性胶质瘤患者术后给予ACNU化疗后外周血淋巴细胞的SCEs。静脉注射ACNU后1 ~ 75 d， SCEs显著升高（从6.3 ~ 9.1/cells增加到17.0 ~ 46.0/cells）。在ACNU后的早期检查中，检测到的sce数量较多。鞘内给药ACNU后，SCEs适度增加至11.7-16.4个/细胞，但放射治疗后没有增加。我们发现，检测外周血淋巴细胞的SCEs可以检测到ACNU治疗后正常宿主细胞的损伤情况，有助于监测ACNU治疗恶性胶质瘤患者的临床疗效。目前正在进行一项研究，以评估我们是否可以通过估计计划接受ACNU治疗的患者淋巴细胞的体外SCEs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.