Atractylenolide I Induces Apoptosis and Suppresses Glycolysis by Blocking the JAK2/STAT3 Signaling Pathway in Colorectal Cancer Cells

Abstract

Colorectal cancer (CRC) is the third most common cancer worldwide and is associated with a poor clinical outcome and survival. Therefore, the development of novel therapeutic agents for CRC is imperative. Atractylenolide I (AT-I) is a sesquiterpenoid lactone derivative of Rhizoma Atractylodis macrocephalae that exhibits diverse biological activities, including anti-cancer activities. However, the effects and potential mechanism of AT-I in CRC have yet to be fully elucidated. In this study, we aimed to examine the anti-cancer properties of AT-I and the associated functional mechanisms in vitro and in vivo. We found that AT-I treatment significantly suppressed the viability of CRC cell lines and inhibited colony formation, but to a lesser extent in NCM460 cells. Annexin V/PI staining showed that AT-I induced apoptosis in CRC cells, accompanied by increased caspase-3 and PARP-1 cleavage, enhanced expression of Bax, and reduced expression of Bcl-2. Furthermore, AT-I blocked cell glycolysis by inhibiting both glucose uptake and lactate production in CRC cells, and specifically downregulated the expression of the rate-limiting glycolytic enzyme HK2. In contrast, it had no discernable effects on the glycolytic enzymes PFK and PKM2. A mechanistic study revealed that AT-1 negatively regulates STAT3 phosphorylation through direct interaction with JAK2, thereby inhibiting its activation. Moreover, restoring the expression of STAT3 reversed the effect of AT-I on apoptosis and glycolysis in CRC cells. In vivo results revealed that AT-I significantly suppressed tumor growth in HCT116-xenografted mice. Collectively, our findings indicate that the anticancer activity of AT-I in CRC is associated with the induction of apoptosis and suppression of glycolysis in CRC cells, via the disruption of JAK2/STAT3 signaling. Our preliminary experimental data indicate that AT-I may have applications as a promising candidate for the treatment of CRC.

Mechanism overview and target mining of atherosclerosis: Endothelial cell injury in atherosclerosis is regulated by glycolysis (Review)

“…In addition, it can enhance cell metabolism through HK2 upregulation. Previous studies have demonstrated that the IL-6/STAT3 and JAK2/STAT3 pathways can promote glycolysis, confirming the crucial role of STAT3 in AS and glycolysis (96,97). Therefore, STAT3 was proven to be an important factor in AS and glycolysis.…”

Atractylenolide-I Suppresses Tumorigenesis of Breast Cancer by Inhibiting Toll-Like Receptor 4-Mediated Nuclear Factor-κB Signaling Pathway

“…AT-I is a novel TLR4-antagonizing agent (Liu et al, 2016), and it exerted anti-tumor effects on colorectal cancer, bladder cancer and melanoma (Fu et al, 2018;Guo et al, 2019;Li et al, 2020), then we investigated whether it could suppress tumorigenesis in Rat breast tissues were kept to detect TLR4/NF-κB pathway by western blot assay at the end of experiment. (G, H, I) TNF-α, IL-6 and IL-1β levels in rat breast tissues were analyzed by ELISA at the end of experiment.…”

Atractylenolide-I suppresses tumorigenesis of breast cancer by inhibiting toll-like receptor 4 (TLR4)-mediated NF-κB signaling pathway

“…AT-I is a novel TLR4-antagonizing agent [20], and it exerted anti-tumor effects on colorectal cancer, bladder cancer and melanoma [17][18][19], then we investigated whether it could suppress tumorigenesis in breast cancer via inhibiting TLR4/NF-κB pathway. Firstly, we found that AT-I could inhibit cell growth, proliferation, migration and invasion, and induce apoptosis in breast cancer cells.…”

“…Rhizoma Atractylodis Macrocephalae, one of the traditional Chinese crude materials, had signi cant gastrointestinal tract protective, neuroprotective and immunomodulatory activities [13], and numerous lines of evidence showed that it also exerted anti-tumor, anti-in ammatory and antioxidant effects [13,14]. Atractylenolide I (AT-I), the major bioactive component from Rhizoma Atractylodes Macrocephalae, also had multiple therapeutic activities including anti-in ammatory [15,16] and anti-tumor effects [17][18][19]. Speci cally, AT-I could ameliorate LPS-induced lung damages in mouse model [15], and suppressed LPS-induced NO release and diminished pro-in ammatory cytokines levels in BV-2 cells [16].…”

“…Speci cally, AT-I could ameliorate LPS-induced lung damages in mouse model [15], and suppressed LPS-induced NO release and diminished pro-in ammatory cytokines levels in BV-2 cells [16]. AT-I had the anti-tumor effects against many cancers, such as colorectal cancer [17], non-small cell lung cancer [18] and melanoma [19], and it also had a binding site similar to LPS and served as a novel TLR4antagonizing agent [20]. Moreover, a randomized pilot study of AT-I on gastric cancer cachexia patients showed that AT-I could inhibit pro-in ammatory cytokines and proteolysis-inducing factor (PIF) proteolysis, and then alleviating symptoms of gastric cancer cachexia patients [21].…”