Reactive oxygen species mediate anlotinib-induced apoptosis via activation of endoplasmic reticulum stress in pancreatic cancer

Abstract

Anlotinib (AL3818), a novel multi-targeted receptor tyrosine kinase inhibitor, has recently been proven to be an antitumour drug. This study aimed to explore the antitumour effect of anlotinib and its underlying molecular mechanisms in human pancreatic cancer (PC) cells. The anti-proliferative effect of anlotinib for three PC cell lines was validated using CCK-8, colony formation and EdU detection assays. Cell cycle, cell apoptosis, and reactive oxygen species (ROS) detection assays, a PC xenograft model and immunohistochemistry were performed to elucidate the mechanisms by which anlotinib induced tumour lethality in vitro and in vivo. These results demonstrated that anlotinib inhibited proliferation, induced G2/M phase arrest and triggered apoptosis in PC cell lines. Anlotinib induced PC’s apoptosis through the accumulation of ROS which activated the endoplasmic reticulum (ER) stress via PERK/p-eIF2α/ATF4 pathway. Furthermore, we demonstrated that the expression level of Nrf2, an antioxidant protein, increased with anlotinib treatment. Nrf2 knockdown enhanced the pro-apoptotic effect of anlotinib and the expression of the PERK/p-eIF2α/ATF4 pathway. The in vivo results suggested that suppressing Nrf2 improved the antitumour effect of anlotinib on PC cells. These data indicated that the apoptotic effect of anlotinib on PC cells was induced by ER stress via the accumulation of ROS. In the future, anlotinib combined with an Nrf2 inhibitor may provide a new therapeutic strategy for the treatment of human PC.

Multi-Omics Analysis of Anlotinib in Pancreatic Cancer and Development of an Anlotinib-Related Prognostic Signature

“…Anlotinib inhibits proliferation and induces apoptosis of pancreatic cancer cells, as shown in this work and in a recent study by Yang L. et al (2020) . In addition, our work also revealed that anlotinib impairs the migration and invasion capability of pancreatic cancer cells ( Figures 1E–H ).…”

“…This phenomenon is observed in this work, as cell cycle assay showed that the ratio of pancreatic cancer cells in G2/M phase was dramatically elevated after anlotinib treatment ( Figures 2D–G ). Transcriptomics analysis has been applied to explore the mechanism of anlotinib in several tumors like intrahepatic cholangiocarcinoma, pancreatic cancer, colon cancer, and synovial sarcoma ( Tang et al, 2019 ; Song et al, 2020 ; Sun et al, 2020 ; Yang L. et al, 2020 ). Yang L. et al (2020) reported that DEGs in pancreatic cancer cells with anlotinib treatment are associated with apoptosis and ER stress, and demonstrated that anlotinib induces apoptosis of pancreatic cancer cells through the activation of ER stress via PERK/p-eIF2α/ATF4 pathway, supporting the findings in this work.…”

“…Transcriptomics analysis has been applied to explore the mechanism of anlotinib in several tumors like intrahepatic cholangiocarcinoma, pancreatic cancer, colon cancer, and synovial sarcoma ( Tang et al, 2019 ; Song et al, 2020 ; Sun et al, 2020 ; Yang L. et al, 2020 ). Yang L. et al (2020) reported that DEGs in pancreatic cancer cells with anlotinib treatment are associated with apoptosis and ER stress, and demonstrated that anlotinib induces apoptosis of pancreatic cancer cells through the activation of ER stress via PERK/p-eIF2α/ATF4 pathway, supporting the findings in this work. KEGG enrichment analysis based on anlotinib induced DEGs in intrahepatic cholangiocarcinoma revealed that cell cycle is the most significantly enriched pathway ( Song et al, 2020 ).…”

Anlotinib: A Novel Targeted Drug for Bone and Soft Tissue Sarcoma

“…Anlotinib was developed by Nanjing Chia Tai Tianqing Pharmaceutical Co., Ltd. as a new oral molecular RTK inhibitor; it targets VEGFR1, VEGFR3, VEGFR2/KDR, PDGFR-a, c-Kit, and FGFRs 1-3 and inhibits TA and tumor cell proliferation (31,(41)(42)(43). Anlotinib may inhibit more targets than that do other RTK inhibitors, such as pazopanib, sunitinib, and sorafenib.…”

In vitro Anticancer Effects of JI017 on Two Prostate Cancer Cell Lines Involve Endoplasmic Reticulum Stress Mediated by Elevated Levels of Reactive Oxygen Species

“…Reactive oxygen species have recently emerged as important regulators of ER function and UPR activation, and ER stress and increased ROS production were detected on chemotherapy treatment in several malignancies ( Cubillos-Ruiz et al, 2017 ; Lin et al, 2019 ; Qi et al, 2020 ; Yang and et al, 2020 ). ROS are an essential component of the signaling that leads to protein misfolding and ER stress-induced cell death ( Verfaillie et al, 2012 ).…”