Antiproliferative Effect of 7-Ketositosterol in Breast and Liver Cancer Cells: Possible Impact on Ceramide, Extracellular Signal-Regulated Kinases, and Nuclear Factor Kappa B Signaling Pathways

Hanumanthu, S.Umarani, K.Anand Kumar, K.Ganesh

» doi: http://https://ijpsl.co.in//.2023.v13.i04.pp33-48

Abstract

7-Ketositosterol (7-KSS) was the focus of this investigation because of its potential effects on sphingomyelin/ceramide metabolites and cell death in HepG2 and MCF-7 human liver cancer cells. Methods: At various concentrations and times, the anti-proliferative effects of 7-KSS therapy were evaluated. Cell viability was determined by MTT analysis, while sphingomyelins (SMs), ceramides (CERs), and sphingosine-1-phosphate (S1P) levels were analyzed by LC-MS/MS. Western blot analysis and immunofluorescence labeling were used to determine the amounts of phosphorylated 44/42 ERK1/2 and NF-?B p65 (Ser536) proteins. The process of cell death was examined by using flow cytometric analysis of annexin-V and propidium iodide (PI) labeling, as well as TUNEL staining. In comparison to control cells, 7- KSS treatment considerably reduced cell viability and levels of S1P, p-44/42 ERK1/2, and pNF-?B p65 proteins in cancer cells. In cancer cells treated with 7-KSS, apoptosis and intracellular levels of C16-C24 CERs were shown to increase significantly. Results: S1P, p44/42 ERK1/2, and p-NF-?B p65 protein levels were downregulated by 7-KSS, which decreased cell proliferation and increased ceramide buildup and apoptosis.