Cucurbitacin D Inhibits the Proliferation of HepG2 Cells and Induces Apoptosis by Modulating JAK/STAT3, PI3K/Akt/mTOR and MAPK Signaling Pathways

Abstract

Background: Cucurbitacin D (CuD) is a natural compound that can be isolated in various plant families, mainly from Ecballium Elaterium (L.) A. Rich. (E. Elaterium). It is a triterpenoid with a broad spectrum of biological activity, including anti-cancer properties. Hepatocellular carcinoma, the aggressive type of liver cancer, is an important public health problem worldwide. Objective: In the present study, we investigated the anticancer effect of CuD treated at different doses on the HepG2 cell line and the underlying mechanism in vitro. Method: CuD was isolated from the fruit juice of E. Elaterium plant and quantitative analysis was performed using high-performance liquid chromatography. The cell viability effect of purified CuD was determined by the MTT test also cell apoptosis and cell cycle arrest effects were determined by flow cytometry. DNA damage was evaluated with the comet test. Proteins and genes involved in PI3K/AKT/mTOR, MAPK, and JAK2/STAT3 signaling pathways were evaluated by western blot and qRT-PCR. Result: CuD showed both antiproliferative and cytotoxic effects against the HepG2 cell line in a dose and time-dependent manner. It was observed that CuD induced apoptosis and blocked the cell cycle in HepG2 cells. It was observed that the expression of genes and some proteins that play a key role in PI3K/AKT/mTOR, MAPK, and JAK2/STAT3 cascades were dose-dependently down-regulated and activated the apoptotic pathway. Conclusion: All these results; show promise that CuD may have a therapeutic effect in liver cancer with HCC.

Investigation of apoptotic effects of Cucurbitacin D, I, and E mediated by Bax/Bcl‐xL, caspase‐3/9, and oxidative stress modulators in HepG2 cell line

“…CuD, CuI, and CuE were prepared as described by Uremis et al (2022a; 2022b). Briefly, the juice of the Ecballium Elaterium plant was obtained and lyophilized.…”

“…Gene expression analysis was performed as in our previous study (Üremiş et al, 2022a; 2022b). Total RNA was procured from cells exposed to CuD, CuI, and CuE using an RNA Isolation Kit (Gene All Hybrid‐R).…”

“…The development of cancer is currently understood to be the result of the activation of signaling pathways that allow cells to proliferate uncontrollably, inhibition of apoptosis due to protein damage and genetic mutations (Sever & Brugge, 2015), and changes occurring at the molecular, cellular, and organismal levels due to oxidative stress, which can trigger the formation of various types of cancer through a cumulative process (Ackerman & Simon, 2014). While recent studies have investigated the link between cucurbitacins and specific signaling pathways in HCC regarding their apoptotic effects (Üremiş et al, 2022a; 2022b), the relationship between cucurbitacins and the caspase cascade, the Bcl‐2 family, and oxidative stress modulators has not yet been fully elucidated. Therefore, in our study, we evaluated the apoptosis‐inducing effects of cucurbitacins D, I, and E (CuD, CuI, and CuE) in the HepG2 cell line, as well as their modulation of the caspase cascade and oxidant damage.…”

Dexpanthenol exhibits antiapoptotic and anti‐inflammatory effects against nicotine‐induced liver damage by modulating Bax/Bcl‐xL, Caspase‐3/9, and Akt/NF‐κB pathways

“…The western blot analysis method, used in previous studies, was applied to determine protein levels. [14,29] Homogenized samples of liver tissue were placed in radioimmunoprecipitation assay lysis buffer and then centrifuged at 20,000g for 10 min. The supernatants were separated, and the levels of protein were determined.…”

“…[12,13] Activated PI3K phosphorylates phosphoinositide-dependent kinase-1 (PDK-1), which further activates Akt. [14] Nicotine-induced Akt activation stimulates the NF-κB pathway, increasing apoptosisrelated proteins and inflammatory responses. [13,15,16] Aberrant expression of these proteins plays a pivotal role in nicotine-induced cell growth, contributing to various diseases.…”