Identification and clinical application of novel antiapoptotic factor produced from ovarian cancer cells

2006年至2007年，我校动物中心发生小鼠肝炎病毒污染事件。包括我们的小鼠在内的所有实验动物首先被杀死，然后对中心进行消毒。然而，消毒6个月后，动物中心再次发现病毒。我们改变了实验计划，尝试更多的体外实验。2007年底，重新启动动物实验，目前正在进行小鼠杂交瘤实验。2007年，第一位首席调查员退休。因此，出乎意料的是，第一个实验计划不能完全执行，现在我们一直在继续原计划。(1) m细胞源性抗凋亡物质（MAAS）的生化表征：从培养上清中部分纯化MAAS，免疫Bal b/c小鼠制备特异性抗体产生杂交瘤。然而，由于动物中心的污染事故，所有免疫小鼠都被杀死了。自动物中心重新开放以来，动物实验已经重新开始。该单克隆抗体的建立将为分离纯化MAAS分子和克隆其cDNA提供可能。(2) MAAS对抗凋亡现象的生物学表征：(1)我们在其他实验中研究了抗肿瘤药物诱导卵巢衰竭的分子机制。偶然的机会，这些实验发现了可能的抗凋亡机制类似于maas诱导的抗凋亡。(2)我们从M细胞中建立了几种抗cddp细胞系。两株抗cddp细胞株的MAAS活性均高于亲本细胞。这些数据表明，MAAS活性的增加可能是获得性CDDP抗性的一个可能机制。

The animal center of our university had been contaminated with mouse hepatitis virus from 2006 to 2007. All the experimental animals including our mice were killed at first, and then the center was disinfected. However, 6 months after the disinfection, virus was found out again in the animal center. We changed experimental plans to try more in vitro experiments. In the end of 2007, animal experiments were restarted and we are now performing mouse hybridoma experiments. In 2007, the first head investigator was retired. Therefore, unexpectedly the first experimental plans could not performed completely and now we have been continuing the original plans.(1) Biochemical characterization of M-cell derived antiapoptotic substances (MAAS): We partially purified MAAS from the culture supernatants and immunized Bal b/c mice to mace specific antibody-producing hybridoma. However, all the immunized mice were killed because of the contamination accident in the animal center. Since reopening of the animal center, animal experiments have been restarted. Establishment of the monoclonal antibody will make possible to isolate pure MAAS molecules and clone their cDNA.(2) Biological characterization of anti-apoptotic phenomena by MAAS: (1) We have been investigating molecular mechanisms of anticancer drug-induced ovarian failure in the other experiments. By chance, those experiments had found possible antiapoptotic mechanisms similar to MAAS-induced antiapoptosis. (2) We have established several CDDP-resistant cell lines from M cells. Two CDDP-resistant cell lines produced much higher MAAS activity than that the parent cells. These data indicate that increased MAAS activity can be a possible mechanism of an acquired CDDP resistance.