GPR68 的抑制和敲除可诱导铁死亡，从而对体外胶质母细胞瘤细胞的存活产生负面影响。在此，我们的目的是使用 GBM 细胞系 U87-MG 和 U138-MG 在斑马鱼中使用两个原位幼虫异种移植模型来证明抑制 GPR68 会降低体内胶质母细胞瘤细胞的存活率。在 GPR68 抑制或敲低的情况下评估癌细胞的体内存活率。在体外，shRNA 介导的 GPR68 抑制敲低显示出对 U87 和 U138 胶质母细胞瘤细胞系的有效细胞毒性作用。这种效应与细胞内脂质过氧化增加有关，表明铁死亡是细胞死亡的潜在机制。将这些发现转化为体内实验，我们通过成功移植荧光标记的人胶质母细胞瘤细胞，在斑马鱼中建立了一种新型异种移植模型，这些细胞先前已被证明过度表达 GPR68。 GPR68 的 shRNA 敲低显着降低了该模型中移植 GBM 细胞的活力。此外，食人吗啡肽（OGM）（一种 GPR68 的高度特异性小分子抑制剂）治疗也降低了移植 GBM 细胞的活力，对发育中的斑马鱼胚胎的毒性有限。这项研究表明，用 OGM 等小分子治疗 GPR68 是治疗 GBM 的一种有前途的方法。© 2024。作者。

Inhibition and knockdown of GPR68 negatively affects glioblastoma cell survival in vitro by inducing ferroptosis. Herein, we aimed to demonstrate that inhibition of GPR68 reduces the survival of glioblastoma cells in vivo using two orthotopic larval xenograft models in Danio rerio, using GBM cell lines U87-MG and U138-MG. In vivo survival of the cancer cells was assessed in the setting of GPR68 inhibition or knockdown.In vitro, shRNA-mediated knockdown of GPR68 inhibition demonstrated potent cytotoxic effects against U87 and U138 glioblastoma cell lines. This effect was associated with increased intracellular lipid peroxidation, suggesting ferroptosis as the underlying mechanism of cell death. Translating these findings in vivo, we established a novel xenograft model in zebrafish by successfully grafting fluorescently labeled human glioblastoma cells, which were previously shown to overexpress GPR68. shRNA knockdown of GPR68 significantly reduced the viability of grafted GBM cells within this model. Additionally, treatment with ogremorphin (OGM), a highly specific small molecule inhibitor of GPR68, also reduced the viability of grafted GBM cells with limited toxicity to the developing zebrafish embryos. This study suggests that therapeutic targeting of GPR68 with small molecules like OGM represents a promising approach for the treatment of GBM.© 2024. The Author(s).