datasheet1_The Widely Used Antihelmintic Drug Albendazole is a Potent Inducer of Loss of Heterozygosity.docx

The antihelmintic drug ABZ and its metabolites belong to the chemical family of benzimidazoles (BZM) that act as potent tubulin polymerization inhibitors, suggesting a potential re-direction of BZMs for cancer therapy. Applying UV-Vis spectrometry we here demonstrate ABZ as a DNA intercalator. This insight led us to determine the primary mode of ABZ action in mammalian cells. As revealed by RNA sequencing, ABZ did neither grossly affect replication as analyzed by survival and replication stress signaling, nor the transcriptome. Actually, unbiased transcriptome analysis revealed a marked cell cycle signature in ABZ exposed cells. Indeed, short-term exposure to ABZ arrested mammalian cells in G2/M cell cycle stages associated with frequent gains and losses of chromatin. Cellular analyses revealed ABZ as a potent mammalian spindle poison for normal and malignant cells, explaining the serious chromosome segregation defects. Since chromosomal aberrations promote both cancer development and cell death, we determined if besides its general cytotoxicity, ABZ could predispose to tumor development. As measured by loss of heterozygosity (LOH) in vitro and in vivo ABZ was found as a potent inducer of LOH and accelerator of chromosomal missegregation.

抗蠕虫药ABZ及其代谢产物属于苯并咪唑类（benzimidazoles, BZM）化合物家族，该类化合物为强效微管蛋白聚合抑制剂，提示BZMs有望被重新定向用于癌症治疗。本研究采用紫外-可见分光光度法（UV-Vis spectrometry）证实，ABZ可作为DNA嵌入剂。这一发现促使我们探究ABZ在哺乳动物细胞中的主要作用机制。经RNA测序（RNA sequencing）分析显示，ABZ既未对通过细胞存活与复制应激信号评估的复制过程造成显著影响，也未对转录组产生明显作用。实际上，无偏倚转录组分析结果表明，经ABZ处理的细胞呈现出显著的细胞周期特征。具体而言，短期暴露于ABZ可使哺乳动物细胞阻滞于G2/M细胞周期时相，该过程伴随染色质的频繁增减。细胞分析显示，ABZ对正常细胞与恶性细胞均为强效的哺乳动物纺锤体毒素，这可解释其引发的严重染色体分离缺陷。由于染色体畸变既可促进癌症发生，又可诱导细胞死亡，因此我们探究了ABZ除具备一般性细胞毒性之外，是否还会诱发肿瘤发生。通过体外与体内的杂合性缺失（loss of heterozygosity, LOH）实验检测发现，ABZ可强效诱导LOH发生，并加速染色体错分离。