PO.ET01.05 · 实验与分子治疗
选择性蛋白激酶 C-iota 抑制剂在抑制 PI3K/AKT 驱动的胰腺导管腺癌增殖中的治疗潜力
Therapeutic potential of a selective protein kinase Ciota inhibitor, in suppressing PI3K/AKT driven pancreatic ductal adenocarcinoma proliferation
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摘要 Abstract
中文摘要
胰腺癌由于其晚期诊断和显著的化疗耐药性，仍然是最致命的恶性肿瘤之一，位居美国癌症相关死亡的第三位。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），是驱动癌细胞增殖、转移和存活的致癌信号通路的关键调节因子。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] 这一选择性 PKC-ι 抑制剂在胰腺腺癌细胞系 AsPC1 和 PANC1 中的治疗潜力。剂量-反应分析显示，在 10 μM ICA-1S 时抑制作用最大，AsPC1 细胞增殖降低 52%，PANC1 细胞降低 49.8%。在 AsPC1 细胞系上进行的 Western blot 分析证实，处理后 PKC-ι 和 PKC-ζ 蛋白表达下调。机制研究表明，ICA-1S（10 μM）显著减弱了磷脂酰肌醇 3-激酶（PI3K）/蛋白激酶 B（AKT）信号级联，使磷酸化磷脂酰肌醇 3-激酶（pPI3K）降低 51%，总 PI3K 降低 18%。这种抑制导致磷酸化 AKT（pAKT）减少 39%，总 AKT 减少 18%，表明下游 PI3K 通路信号传导受到抑制。此外，ICA-1S 处理诱导了 AsPC1 细胞的凋亡，表现为切割型 Caspase-9 的表达增加 10%、细胞色素 c 释放增加 17%，以及抗凋亡蛋白 Survivin 的表达降低 42%。划痕愈合实验进一步证明了 ICA-1S 的抗转移作用，处理后的 AsPC1 细胞即使在五天后仍无法闭合划痕，而对照细胞实现了完全闭合。总体而言，这些发现表明 ICA-1S 有效抑制了 PKC-ι 介导的 PI3K/AKT 信号传导并促进胰腺癌细胞凋亡，凸显了其作为胰腺腺癌有前景的靶向治疗药物的潜力。正在进行的研究将采用水溶性四唑盐（WST）实验、免疫沉淀、免疫荧光和扩展的 Western blot 分析，以进一步阐明 ICA-1S 的下游分子效应。
查看英文原文 English abstract
Pancreatic cancer remains one of the most lethal malignancies due to its late-stage diagnosis and pronounced chemoresistance, ranking as the third leading cause of cancer-related deaths in the United States. Atypical Protein Kinase C (aPKC) isoforms, Protein Kinase C-iota (PKC-ι) and Protein Kinase C- zeta (PKC-ζ), are critical regulators of oncogenic signaling pathways that drive cancer cell proliferation, metastasis, and survival. In this study, we investigated the therapeutic potential of ICA-1S [5-amino-1-((1R,2S,3S,4R)-2,3-dihydroxy-4-methylcyclopentyl)-1H-imidazole-4-carboxamide], a selective PKC-ι inhibitor, in pancreatic adenocarcinoma cell lines AsPC1 and PANC1. Dose-response analyses revealed maximal inhibitory effects at 10 μM ICA-1S, reducing cell proliferation by 52% in AsPC1 and 49.8% in PANC1 cells. Western blot analysis performed on AsPC1 cell line confirmed downregulation of PKC-ι and PKC-ζ protein expression following treatment. Mechanistic studies demonstrated that ICA-1S (10 μM) significantly attenuated the Phosphatidylinositol 3-Kinase (PI3K)/Protein Kinase B (AKT) signaling cascade, phosphorylated Phosphatidylinositol 3-Kinase (pPI3K) by 51%, and total PI3K by 18%. This suppression led to a 39% decrease in phosphorylated AKT (pAKT) and an 18% decrease in total AKT, demonstrating inhibition of downstream PI3K pathway signaling. Additionally, ICA-1S treatment induced apoptosis in AsPC1 cells, as evidenced by increased expression of Cleaved Caspase-9 by 10%, increased Cytochrome c release by 17%, and decreased expression of the anti-apoptotic protein Survivin by 42%. Wound-healing assays further demonstrated the anti-metastatic effect of ICA-1S, with treated AsPC1 cells failing to close wounds even after five days, while control cells achieved complete closure.Collectively, these findings indicate that ICA-1S effectively suppresses PKC-ι-mediated PI3K/AKT signaling and promotes apoptosis in pancreatic cancer cells, underscoring its potential as a promising targeted therapeutic agent for pancreatic adenocarcinoma. Ongoing studies will employ Water-Soluble Tetrazolium (WST) assays, immunoprecipitation, immunofluorescence, and extended Western blot analyses to further delineate the downstream molecular effects of ICA-1S.
利益披露 Disclosure
S. Rimal, None..
G. R. Khanal, None.