Department of General Surgery, Qinghai Provincial People's Hospital, Xining 810007, Qinghai Province, China.
Department of Clinical Pharmacy, The Affiliated Hospital of Qinghai University, Xining 810001, Qinghai Province, China.
World J Gastroenterol. 2024 Jun 7;30(21):2793-2816. doi: 10.3748/wjg.v30.i21.2793.
Pancreatic cancer (PC) is associated with some of the worst prognoses of all major cancers. Thymoquinone (TQ) has a long history in traditional medical practice and is known for its anti-cancer, anti-inflammatory, anti-fibrosis and antioxidant pharmacological activities. Recent studies on hypoxia-inducible factor-1α (HIF-1α) and PC have shown that HIF-1α affects the occurrence and development of PC in many aspects. In addition, TQ could inhibit the development of renal cancer by decreasing the expression of HIF-1α. Therefore, we speculate whether TQ affects HIF-1α expression in PC cells and explore the mechanism.
To elucidate the effect of TQ in PC cells and the regulatory mechanism of HIF-1α expression.
Cell counting kit-8 assay, Transwell assay and flow cytometry were performed to detect the effects of TQ on the proliferative activity, migration and invasion ability and apoptosis of PANC-1 cells and normal pancreatic duct epithelial (hTERT-HPNE) cells. Quantitative real-time polymerase chain reaction and western blot assay were performed to detect the expression of HIF-1α mRNA and protein in PC cells. The effects of TQ on the HIF-1α protein initial expression pathway and ubiquitination degradation in PANC-1 cells were examined by western blot assay and co-immunoprecipitation.
TQ significantly inhibited proliferative activity, migration, and invasion ability and promoted apoptosis of PANC-1 cells; however, no significant effects on hTERT-HPNE cells were observed. TQ significantly reduced the mRNA and protein expression levels of HIF-1α in PANC-1, AsPC-1, and BxPC-3 cells. TQ significantly inhibited the expression of the HIF-1α initial expression pathway (PI3K/AKT/mTOR) related proteins, and promoted the ubiquitination degradation of the HIF-1α protein in PANC-1 cells. TQ had no effect on the hydroxylation and von Hippel Lindau protein mediated ubiquitination degradation of the HIF-1α protein but affected the stability of the HIF-1α protein by inhibiting the interaction between HIF-1α and HSP90, thus promoting its ubiquitination degradation.
The regulatory mechanism of TQ on HIF-1α protein expression in PC cells was mainly to promote the ubiquitination degradation of the HIF-1α protein by inhibiting the interaction between HIF-1α and HSP90; Secondly, TQ reduced the initial expression of HIF-1α protein by inhibiting the PI3K/AKT/mTOR pathway.
胰腺癌（PC）的预后是所有主要癌症中最差的之一。姜黄素（TQ）在传统医学实践中有着悠久的历史，其具有抗癌、抗炎、抗纤维化和抗氧化的药理学活性。最近关于缺氧诱导因子-1α（HIF-1α）和 PC 的研究表明，HIF-1α 通过多种方式影响 PC 的发生和发展。此外，TQ 可以通过降低 HIF-1α 的表达来抑制肾癌的发展。因此，我们推测 TQ 是否会影响 PC 细胞中 HIF-1α 的表达，并探讨其机制。
阐明 TQ 在 PC 细胞中的作用及调控 HIF-1α 表达的机制。
通过细胞计数试剂盒-8 检测、Transwell 检测和流式细胞术检测 TQ 对 PANC-1 细胞和正常胰腺导管上皮（hTERT-HPNE）细胞增殖活性、迁移和侵袭能力及凋亡的影响。实时定量聚合酶链反应和 Western blot 检测 PC 细胞中 HIF-1α mRNA 和蛋白的表达。Western blot 和免疫共沉淀检测 TQ 对 PANC-1 细胞中 HIF-1α 蛋白初始表达途径和泛素化降解的影响。
TQ 显著抑制 PANC-1 细胞的增殖活性、迁移和侵袭能力，并促进其凋亡；而对 hTERT-HPNE 细胞无明显作用。TQ 显著降低 PANC-1、AsPC-1 和 BxPC-3 细胞中 HIF-1α 的 mRNA 和蛋白表达水平。TQ 显著抑制 HIF-1α 初始表达途径（PI3K/AKT/mTOR）相关蛋白的表达，并促进 PANC-1 细胞中 HIF-1α 蛋白的泛素化降解。TQ 对 HIF-1α 蛋白的羟化和 von Hippel Lindau 蛋白介导的泛素化降解无影响，但通过抑制 HIF-1α 与 HSP90 的相互作用影响 HIF-1α 蛋白的稳定性，从而促进其泛素化降解。
TQ 调节 PC 细胞中 HIF-1α 蛋白表达的机制主要是通过抑制 HIF-1α 与 HSP90 的相互作用促进 HIF-1α 蛋白的泛素化降解；其次，TQ 通过抑制 PI3K/AKT/mTOR 通路降低 HIF-1α 蛋白的初始表达。
