杨树菇菌素——一种源自杨树菇的核糖体毒素样蛋白，可增强胶质母细胞瘤原代细胞对常规替莫唑胺化疗的敏感性。

Ageritin-The Ribotoxin-like Protein from Poplar Mushroom () Sensitizes Primary Glioblastoma Cells to Conventional Temozolomide Chemotherapy.

机构信息

IRCCS Istituto Neurologico Mediterraneo NEUROMED, Via Atinense 18, 86077 Pozzilli, Italy.

Department of Environmental, Biological and Pharmaceutical Sciences and Technologies (DiSTABiF), University of Campania "Luigi Vanvitelli", Via Vivaldi 43, 81100 Caserta, Italy.

出版信息

Molecules. 2022 Apr 7;27(8):2385. doi: 10.3390/molecules27082385.

Here, we propose Ageritin, the prototype of the ribotoxin-like protein family, as an adjuvant treatment to control the growth of NULU and ZAR, two primary human glioblastoma cell lines, which exhibit a pharmacoresistance phenotype. Ageritin is able to inhibit NULU and ZAR growth with an IC50 of 0.53 ± 0.29 µM and 0.42 ± 0.49 µM, respectively. In this study, Ageritin treatment highlighted a macroscopic genotoxic response through the formation of micronuclei, which represents the morphological manifestation of genomic chaos induced by this toxin. DNA damage was not associated with either the deregulation of DNA repair enzymes (i.e., ATM and DNA-PK), as demonstrated by quantitative PCR, or reactive oxygen species. Indeed, the pretreatment of the most responsive cell line ZAR with the ROS scavenger N-acetylcysteine (NAC) did not follow the reverse cytotoxic effect of Ageritin, suggesting that this protein is not involved in cellular oxidative stress. Vice versa, Ageritin pretreatment strongly enhanced the sensitivity to temozolomide (TMZ) and inhibited MGMT protein expression, restoring the sensitivity to temozolomide. Overall, Ageritin could be considered as a possible innovative glioblastoma treatment, directly damaging DNA and downregulating the MGMT DNA repair protein. Finally, we verified the proteolysis susceptibility of Ageritin using an in vitro digestion system, and considered the future perspective use of this toxin as a bioconjugate in biomedicine.

在这里，我们提出了一种名为 Ageritin 的核糖核酸酶样蛋白家族原型，作为一种辅助治疗方法，以控制 NULU 和 ZAR 两种原发性人胶质母细胞瘤细胞系的生长，这两种细胞系表现出药物抵抗表型。Ageritin 能够以 0.53 ± 0.29 µM 和 0.42 ± 0.49 µM 的 IC50 分别抑制 NULU 和 ZAR 的生长。在这项研究中，Ageritin 处理通过形成微核突出显示了宏观遗传毒性反应，微核是这种毒素诱导的基因组混乱的形态表现。通过定量 PCR 或活性氧（ROS），DNA 损伤与 DNA 修复酶（如 ATM 和 DNA-PK）的失调无关。实际上，用 ROS 清除剂 N-乙酰半胱氨酸（NAC）预处理最敏感的细胞系 ZAR，并没有遵循 Ageritin 的相反细胞毒性作用，这表明该蛋白不参与细胞氧化应激。相反，Ageritin 预处理强烈增强了对替莫唑胺（TMZ）的敏感性，并抑制了 MGMT 蛋白表达，恢复了对替莫唑胺的敏感性。总的来说，Ageritin 可以被认为是一种潜在的胶质母细胞瘤创新治疗方法，它直接损伤 DNA 并下调 MGMT DNA 修复蛋白。最后，我们使用体外消化系统验证了 Ageritin 的蛋白水解易感性，并考虑了未来将这种毒素用作生物医学中生物缀合物的应用前景。