据报道，前列脯氨酸羟化酶亚基alpha 1（P4HA1）促进了胶质瘤的肿瘤生长和转移；因此，针对P4HA1可能是治疗胶质瘤的一种有前途的治疗策略。考虑到体内小干扰RNA（siRNA）的不稳定性，我们采用了载有P4HA1 siRNA的壳聚糖明胶微球（P4HA1 siRNA@CGM）。首先，进行了凝胶电泳和溶血试验，评估了P4HA1 siRNA@CGM的稳定性和血液相容性。然后，进行了甲基噻唑酮（MTT）实验、细胞集落形成实验、Transwell实验、伤口愈合实验、胶瘤球形成实验、管状形成实验和Western blot实验，评估了P4HA1 siRNA@CGM对胶质瘤生物学功能的影响。最后，将125I标记的P4HA1 siRNA@CGM注入移植小鼠体内，记录放射性核素成像结果，进行Ki67和末端脱氧核苷酸转移酶介导的末端标记（TUNEL）染色，评估P4HA1 siRNA@CGM对胶质瘤生长和凋亡的影响。结果显示，P4HA1 siRNA和P4HA1 siRNA@CGM不仅显著抑制了胶质瘤细胞的增殖、转移、胶瘤球形成以及间质标记物（N-连环蛋白和维门蛋白）的蛋白水平和上皮间质转化的转录因子（Snail、Slug和Twist1）的表达，还抑制了人脑微血管内皮细胞（HBMECs）的管状形成，而P4HA1 siRNA@CGM的抑制效果优于P4HA1 siRNA。以上结果表明P4HA1 siRNA@CGM在胶质瘤的临床治疗中具有可行性。© 2023. 控释协会。

It has been reported that prolyl 4-hydroxylase subunit alpha 1 (P4HA1) promoted tumor growth and metastasis of glioma; thus, targeting P4HA1 may be a promising therapeutic strategy against glioma. In consideration of the instability of siRNA in vivo, the chitosan-gelatin microspheres loaded with P4HA1 siRNA (P4HA1 siRNA@CGM) were employed. Firstly, the gel electrophoresis and hemolytic test were performed to assess the stability and blood compatibility of P4HA1 siRNA@CGM. Then, methyl thiazolyl tetrazolium (MTT), cell colony formation, Transwell assay, wound healing assay, gliosphere formation, tube formation, and Western blot were performed to assess the effects of P4HA1 siRNA@CGM on the biological functions of glioma. Finally, 125I-labeled P4HA1 siRNA@CGM was injected into the xenograft mice, radionuclide imaging was recorded, Ki67 and terminal deoxynucleoitidyl transferase-mediated nick end labeling (TUNEL) staining was performed to assess the effects of P4HA1 siRNA@CGM on tumor growth and apoptosis of glioma in vivo. The results showed that P4HA1 siRNA and P4HA1 siRNA@CGM not only markedly inhibited the proliferation, metastasis, gliosphere formation, and the protein levels of interstitial markers (N-cadherin and vimentin) and the transcription factors of epithelial-mesenchymal transition (EMT) (Snail, Slug, and Twist1) in glioma cells, but also inhibited the tube formation in human brain microvascular endothelial cells (HBMECs), and P4HA1 siRNA@CGM exhibited the better inhibitory effects than P4HA1 siRNA. Above results suggested the feasibility of P4HA1 siRNA@CGM in the clinical treatment of glioma.© 2023. Controlled Release Society.