Department of Breast Surgery, the First Affiliated Hospital of Wenzhou Medical University, Wenzhou 325000, China.
Department of Hepatopancreatobiliary Surgery, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou 325000, China.
Phytomedicine. 2022 Jan;95:153861. doi: 10.1016/j.phymed.2021.153861. Epub 2021 Nov 20.
Rosmarinic acid (RA) has been shown to exert anti-tumor effects on various types of cancer. However, its roles in the treatment of pancreatic ductal adenocarcinoma (PDAC) and the underlying mechanisms remain elusive.
The present study aimed to investigate the therapeutic effects of RA on PDAC as well as the underlying mechanisms.
Evaluation of the effects of RA on PDAC malignancy both in vitro and in vivo.
Cell counting kit 8 (CCK8) assay, colony formation assay, 5-Ethynyl-2'-deoxyuridine (EDU) incorporation assay, cell cycle analysis, and apoptosis assay were conducted to assess the inhibitory effect of RA on PDAC cell proliferation. Meanwhile, western blotting and RT-qPCR assay were performed to detect the target gene expression at protein and mRNA levels, respectively. Moreover, the in vivo anti-tumor activities of RA were assayed in an xenograft mouse model of PDAC.
RA dramatically down-regulated Gli1 and its downstream targets. Further studies showed that RA prevents the nuclear translocation of Gli1, while promoting the degradation of cytosolic Gli1 via the proteasome pathway. Moreover, we observed that RA induced G1/S cell cycle arrest and apoptosis in the PDAC cells through regulating the expression of P21, P27, CDK2, Cyclin E, Bax, and Bcl-2, it inhibited the PDAC cell migration and invasion via E-cadherin and MMP-9. Notably, Gli1 overexpression markedly reversed the above RA-induced effects on PDAC cells, whereas Gli1 knockdown enhanced the effects. Additionally, the in vivo assays demonstrated that RA suppresses the tumor growth of PDAC presumably by inhibiting Gli1.
We provided evidence that RA restrained the nuclear translocation of Gli1 and facilitates Gli1 degradation via proteasome pathway, reducing the malignancy of PDAC cells. These findings implicated RA as a therapeutic agent for PDAC.
迷迭香酸（RA）已被证明对多种类型的癌症具有抗肿瘤作用。然而，其在胰腺导管腺癌（PDAC）治疗中的作用及其潜在机制仍不清楚。
本研究旨在探讨 RA 对 PDAC 的治疗作用及其潜在机制。
评估 RA 对 PDAC 恶性肿瘤的体内外作用。
通过细胞计数试剂盒 8（CCK8）测定、集落形成测定、5-乙炔基-2'-脱氧尿苷（EDU）掺入测定、细胞周期分析和凋亡测定来评估 RA 对 PDAC 细胞增殖的抑制作用。同时，通过蛋白质印迹和 RT-qPCR 检测分别检测目标基因在蛋白和 mRNA 水平的表达。此外，在 PDAC 的异种移植小鼠模型中检测 RA 的体内抗肿瘤活性。
RA 显著下调 Gli1 及其下游靶基因。进一步的研究表明，RA 通过蛋白酶体途径阻止 Gli1 的核转位，同时促进细胞质中 Gli1 的降解。此外，我们观察到 RA 通过调节 P21、P27、CDK2、Cyclin E、Bax 和 Bcl-2 的表达，诱导 PDAC 细胞 G1/S 细胞周期停滞和凋亡，通过 E-钙粘蛋白和 MMP-9 抑制 PDAC 细胞迁移和侵袭。值得注意的是，Gli1 过表达显著逆转了 RA 对 PDAC 细胞的上述诱导作用，而 Gli1 敲低增强了这种作用。此外，体内实验表明，RA 通过抑制 Gli1 抑制 PDAC 的肿瘤生长。
我们提供的证据表明，RA 通过蛋白酶体途径抑制 Gli1 的核转位并促进 Gli1 的降解，从而降低 PDAC 细胞的恶性程度。这些发现表明 RA 可能是 PDAC 的治疗剂。
