PO.ET01.05 · 实验与分子治疗
利用非典型蛋白激酶 C 抑制剂抑制儿童胶质母细胞瘤的增殖
Inhibition in the proliferation of pediatric glioblastoma facilitated with atypical protein kinase c inhibitors
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
儿童胶质母细胞瘤（pGBM）是一种侵袭性脑肿瘤，五年生存率约为 5%。目前的治疗选择有限，主要是由于现有药物的血脑屏障穿透性差。非典型蛋白激酶 C（aPKC）亚型，即蛋白激酶 C-iota（PKC-ι）和蛋白激酶 C-zeta（PKC-ζ），在肿瘤细胞增殖和存活中发挥关键作用。在本研究中，我们研究了 ICA-1S [5-amino-1-((1R,2S,3S,4R)-2,3-dihydroxy-4-methylcyclopentyl)-1H-imidazole-4-carboxamide]（一种选择性 PKC-ι 抑制剂）和 ζ-Stat [8-hydroxy-1,3,6-naphthalenetrisulfonic acid]（一种 PKC-ζ 抑制剂）对儿童胶质母细胞瘤 SF188 细胞的影响。将 SF188 细胞（50,000/孔）接种于 6 孔板中，并用不同浓度的 ICA-1S 和 ζ-Stat 处理三天。在第四天测定细胞活力以生成剂量-反应曲线。ICA-1S 在 75 μM 时显示最大抑制作用，而 ζ-Stat 在 20 μM 时使增殖降低约 50%。这些结果表明 PKC-ι 和 PKC-ζ 均参与 SF188 细胞的增殖。为探究这种抑制背后的分子机制，通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）和 Western blotting 分析了处理和未处理细胞的蛋白裂解物。与对照样品相比，ICA-1S 和 ζ-Stat 处理均导致 PKC-ι 和 PKC-ζ 蛋白表达下调。此外，发现凋亡标志物 Caspase-3 的表达在药物处理的细胞中降低，提示在抑制 aPKC 后凋亡信号传导被激活。这些发现表明，选择性抑制 aPKC 可抑制儿童胶质母细胞瘤细胞的增殖并诱导凋亡。正在进行和未来的研究将采用免疫沉淀、免疫荧光和 WST 实验来阐明下游信号通路，并验证 ICA-1S 和 ζ-Stat 作为儿童胶质母细胞瘤潜在治疗候选药物的价值。
查看英文原文 English abstract
Pediatric glioblastoma (pGBM) is an aggressive brain tumor with a five-year survival rate of approximately 5%. Current therapeutic options are limited, primarily due to poor blood-brain barrier penetration of available drugs. Atypical Protein Kinase C (aPKC) isoforms, Protein Kinase C- iota (PKC-ι) and Protein Kinase C- zeta (PKC-ζ), play crucial roles in tumor cell proliferation and survival. In this study, we investigated the effects of ICA-1S [5-amino-1-((1R,2S,3S,4R)-2,3-dihydroxy-4-methylcyclopentyl)-1H-imidazole-4-carboxamide], a selective PKC-ι inhibitor, and ζ-Stat [8-hydroxy-1,3,6-naphthalenetrisulfonic acid], a PKC-ζ inhibitor, on pediatric glioblastoma SF188 cells. SF188 cells (50,000/well) were seeded in 6-well plates and treated with varying concentrations of ICA-1S and ζ-Stat for three days. On the fourth day, cell viability was determined to generate a dose-response curve. ICA-1S showed maximal inhibition at 75 μM, while ζ-Stat decreased proliferation by approximately 50% at 20 μM. These results suggest that both PKC-ι and PKC-ζ contribute to SF188 cell proliferation. To explore the molecular mechanisms underlying this inhibition, protein lysates from treated and untreated cells were analyzed via Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis (SDS-PAGE) and Western blotting. Both ICA-1S and ζ-Stat treatments resulted in downregulation of PKC-ι and PKC-ζ protein expression compared to control samples. Furthermore, expression of the apoptotic marker Caspase-3 was found to be decreased in drug-treated cells, suggesting activation of apoptotic signaling following inhibition of aPKCs. These findings indicate that selective inhibition of aPKCs suppresses proliferation and induces apoptosis in pediatric glioblastoma cells. Ongoing and future studies will employ immunoprecipitation, immunofluorescence, and WST assays to elucidate downstream signaling pathways and validate ICA-1S and ζ-Stat as potential therapeutic candidates for pediatric glioblastoma.
利益披露 Disclosure
S. Rimal, None.