Methylwogonin exerts anticancer effects in A375 human malignant melanoma cells through apoptosis induction, DNA damage, cell invasion inhibition and downregulation of the mTOR/PI3K/Akt signalling pathway

Abstract

Introduction The main purpose of the present research was to study the anticancer effects of methylwogonin in A375 human malignant melanoma cells by evaluating its effects on apoptosis, DNA fragmentation, cancer cell invasion and the mTOR/PI3K/AKT signalling pathway. Material and methods Effects on cell cytotoxicity were evaluated by MTT assay while a clonogenic assay determined the effects of methylwogonin on colony formation. Fluorescence microscopy evaluated apoptotic effects of methylwogonin in these cells using acridine orange/propidium iodide and Hoechst 33342 staining dyes. Gel electrophoresis evaluated the effects of methylwogonin on DNA fragmentation while the Matrigel invasion assay evaluated the effects of the drug on cancer cell invasion. Effects of methylwogonin on the mTOR/PI3K/AKT signalling pathway were evaluated by western blot assay. Results Methylwogonin induces concentration-dependent as well as time-dependent growth inhibitory effects inducing significant cytotoxicity in these cancer cells. Methylwogonin led to dose-dependent inhibition of colony formation in A375 human malignant melanoma cells. The number of cell colonies decreased significantly as the methylwogonin dose increased from 0, 50, 150, to 300 μM. Methylwogonin treatment of cells at lower doses led to yellow fluorescence (early apoptosis), which changed to red/orange fluorescence, indicating late apoptosis at higher doses. Similar results were obtained using Hoechst 33342 staining, revealing that 50, 150 and 300 μM doses of methylwogonin led to significant morphological changes including chromatin condensation, fragmented nuclei and cellular shrinkage. DNA ladder formation was also observed, and this effect increased with increasing doses of methylwogonin. Methylwogonin also inhibited cancer cell invasion in a dose-dependent manner. Conclusions Different doses of methylwogonin led to concentration-dependent downregulation of phosphorylated PI3K, AKT and mTOR.

Polyphyllin I Promoted Melanoma Cells Autophagy and Apoptosis via PI3K/Akt/mTOR Signaling Pathway

“…Subsequently, we researched the changes of these key proteins associated with PI3K/Akt/mTOR signaling pathway in A375 cells after Polyphyllin I treatment, the results manifested that PI3K/Akt/mTOR signaling pathway was inhibited, the expressions of p-PI3K, p-Akt and p-mTOR was distinctly decreased, but the total protein levels of PI3K, Akt, and mTOR had no significant difference in melanoma A375 cells. To our knowledge, the activated PI3K/Akt/mTOR signaling pathway is able to suppress melanoma cell autophagy and apoptosis [ 26 , 27 ]. Therefore, we considered that the inhibitory effect of Polyphyllin I on the PI3K/Akt/mTOR signaling pathway played indispensable roles in suppressing melanoma growth.…”

Wogonin and its analogs for the prevention and treatment of cancer: A systematic review

“…Furthermore, a study on in vivo melanoma model proposed that upon treatment with WOG, it significantly reduced invasion of the cancer cells (Zhao et al, 2014). Methyl‐WOG exhibited potent anticancer activities against A375 melanoma cells through induction of apoptosis, increased DNA damage, inhibition of cellular invasion, and suppression of the mTOR/PI3K/Akt signalling pathway (Chen et al, 2019). Overall, in case of melanoma, the NF‐κB/ERK/PI3K/Akt/mTOR signaling pathway was mainly modulated by WOG to employ its anticancer activities.…”

Phytochemical Profile and Anticancer Activity from Medicinal Plants Against Melanoma Skin Cancer: A Review

“…It also provides a proapoptotic effect by causing significant morphological changes, including chromatin condensation, fragmented nuclei, and cellular shrinkage. In addition, it also has an antimigration and anti-invasion effect through the mTOR/PI3K/AKT signaling pathway by downregulating PI3K, AKT, and phosphorylated mTOR in a dose-dependent manner (Chen et al, 2019b;He et al, 2021). The activity compounds derivated from S. baicalensis have not been explored yet against other melanoma skin cancer.…”