Busulfan is an alkylating drug alkylates DNA, with an IC50 value of 77μM for human serum Paraoxonase 1[1]. Busulfan has been widely used in cellular and animal models to induce rapidly dividing cell death[2].
In vitro, Busulfan treatment in P39 cells at 100μg/ml for 8 hours induced apoptosis, accompanied by caspase activation and cleavage of Bcl-2 and PARP proteins[3]. Treatment of HepaRG cells with 1000μM Busulfan for 4 days resulted in cellular mitochondrial dysfunction, elevated neutral lipid levels, increased fatty acid uptake, and decreased secretion of very-low-density lipoprotein[4]. Treatment of U2OS and MG-63 cells with 150μM Busulfan for 48 hours significantly inhibited cell viability, up-regulated the miR-200 family and regulated related target genes[5]. Treatment with 100μM Busulfan for 48 hours inhibited autophagy in mouse spermatogonial progenitor cells, resulting in a significant up-regulation of mTOR phosphorylation at Ser2448[6].
In vivo, Busulfan treatment via a single intraperitoneal injection at the dose of 40mg/kg for 56 days caused male C3H/Kam mice permanently infertile and led to the morphological damage to sperm produced by surviving stem cell spermatogonia[7]. In NOD/ LtSz-scid /IL2Rγ knockout mice, two intraperitoneal injections of 25mg/kg Busulfan at 48 and 24 hours before infusion of human cells promoted engraftment of human hematopoietic stem cell (HSC) and significantly improved survival[8].
References:
[1] Söyüt H. Investigation of inhibition of busulfan (Chemotherapeutic Drug) on human serum paraoxonase-1 (PON1)[J]. Int. J. Pharmacol. 2021, 17(8), 572–576.
[2] Berger D P, Winterhalter B R, Dengler W A, et al. Preclinical activity off hepsulfam and busulfan in solid human tumor xenografts and human bone marrow[J]. Anti-Cancer Drugs, 1992, 3(5): 531-540.
[3] Hassan Z, Hassan M, Hellström-Lindberg E. The pharmacodynamic effect of busulfan in the P39 myeloid cell line in vitro[J]. Leukemia, 2001, 15(8): 1240-1247.
[4] Allard J, Bucher S, Ferron P J, et al. Busulfan induces steatosis in HepaRG cells but not in primary human hepatocytes: Possible explanations and implication for the prediction of drug‐induced liver injury[J]. Fundamental & Clinical Pharmacology, 2024, 38(1): 152-167.
[5] Mei Q, Li F, Quan H, et al. Busulfan inhibits growth of human osteosarcoma through miR‐200 family micro RNA s in vitro and in vivo[J]. Cancer science, 2014, 105(7): 755-762.
[6] Wei R, Zhang X, Cai Y, et al. Busulfan suppresses autophagy in mouse spermatogonial progenitor cells via mTOR of AKT and p53 signaling pathways[J]. Stem Cell Reviews and Reports, 2020, 16(6): 1242-1255.
[7] Bucci L R, Meistrich M L. Effects of busulfan on murine spermatogenesis: cytotoxicity, sterility, sperm abnormalities, and dominant lethal mutations[J]. Mutation Research/Fundamental and Molecular Mechanisms of Mutagenesis, 1987, 176(2): 259-268.
[8] Hayakawa J, Hsieh M M, Uchida N, et al. Busulfan produces efficient human cell engraftment in NOD/LtSz-Scid IL2Rγ null mice[J]. Stem cells, 2009, 27(1): 175-182.
Busulfan是一种烷化剂，可通过烷基化作用修饰DNA，对human serum Paraoxonase 1的IC50值为77μM[1]。Busulfan广泛应用于细胞和动物模型中诱导快速分裂细胞死亡[2]。
在体外，100μg/ml的Busulfan处理P39细胞8小时可诱导细胞凋亡，伴随caspase激活及Bcl-2和PARP蛋白裂解[3]。1000μM的Busulfan处理HepaRG细胞4天会导致细胞线粒体功能障碍、中性脂质水平升高、脂肪酸摄取增加及极低密度脂蛋白分泌减少[4]。150μM的Busulfan处理U2OS和MG-63细胞48小时能显著抑制细胞活力，上调miR-200家族并调控相关靶基因[5]。100μM的Busulfan处理小鼠精原祖细胞48小时可抑制自噬过程，显著增强mTOR Ser2448位点磷酸化[6]。
在体内，Busulfan以40mg/kg的剂量单次腹腔注射（56天后）导致雄性C3H/Kam小鼠永久性不育，并导致存活的干细胞精原细胞产生的精子发生形态损伤[7]。NOD/LtSz-scid/IL2Rγ敲除小鼠在输注人细胞前48小时和24小时两次腹腔注射25mg/kg剂量的Busulfan，可促进人造血干细胞（HSC）植入并显著提高生存率[8]。