Cucurbitacin I exhibits anticancer efficacy through induction of apoptosis and modulation of JAK / STAT3 , MAPK / ERK , and AKT / mTOR signaling pathways in HepG2 cell line

Abstract

Hepatocellular carcinoma is a common cancer type, especially among men. Although cucurbitacin I (CuI), widely found in plants belonging to the Ecballium elaterium (E. L) plant family, has been shown to have antitumorigenic properties in many cancer types, its anticancer effect, molecular mechanism, and apoptotic effect mediated by signal pathways on hepatocellular carcinoma have not been fully clarified. In the present study, we investigated the anticancer effect of CuI treated at different doses on the HepG2 cell line and the underlying mechanism in vitro. High-purity CuI was obtained from the E. elaterium plant with the aid of HPLC. The effects of this substance on the viability of cells were studied by the MTT assay. The effects of CuI on cell cycle progression and apoptosis were studied with flow cytometry. DNA breaks were analyzed by the Comet assay method.The proteins and genes involved in the JAK/STAT3, MAPK/ERK, and AKT/mTOR signaling pathways were investigated using Western blot and qRT-PCR, respectively. The results of this study demonstrated that CuI significantly reduced HepG2 cell growth in vitro, induced antiproliferation, and G2/M phase of the cell cycle was interrupted. Practical applicationsCuI administration was shown to downregulate the levels of proteins in the PI3K/ AKT/mTOR, MAPK, and JAK2/STAT3 cascades in HepG2 cells. CuI also reduced the expression of MAPK, STAT3, mTOR, JAK2, and Akt genes in different concentrations.DNA breaks are formed as a result of this effect. CuI, by reducing cell proliferation and promoting apoptosis, was found to have potential as a chemotherapeutic agent of hepatocellular carcinoma.

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).…”

“…While cucurbitacins B and D are present in the highest concentrations in plants of the Cucurbitaceae family, cucurbitacins B, D, E, and I have been the most extensively studied for their anticancer, antioxidant, and anti‐inflammatory activities (Chung et al, 2015). Cucurbitacins have been stated to possess antitumor activity through a range of processes, for instance, inhibiting cell replication (Sikander et al, 2019), halting the cell cycle (Uremis et al, 2022b), inducing apoptosis (Üremiş et al, 2022b), restraining angiogenesis and metastasis (Hsu et al, 2019), and destruction of the protein skeleton and autophagy (Song et al, 2020).…”

“…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.…”