Farnesiferol C Exerts Antiproliferative Effects on Hepatocellular Carcinoma HepG2 Cells by Instigating ROS-Dependent Apoptotic Pathway

Abstract

Farnesiferol C (Far-C) is a coumarin commonly extracted from Ferula asafetida and is popularly used as a traditional source of natural remedy. Liver cancer or hepatocellular carcinoma (HCC) has emerged as a major cause behind cancer burden, and limited therapeutic interventions have further aggravated the clinical management of HCC. In the present study, the authors tested the hypothesis that Far-C-instigated oxidative stress resulted in anti-proliferation and apoptosis instigation within human liver cancer HepG2 cells. The observations reported herewith indicated that Far-C exerted considerable cytotoxic effects on HepG2 cells by reducing the cell viability (p < 0.001) in a dose-dependent manner. Far-C exposure also resulted in enhanced ROS production (p < 0.01) which subsequently led to loss of mitochondrial membrane potential. Far-C-instigated oxidative stress also led to enhanced nuclear fragmentation and condensation as revealed through Hoechst-33342. These molecular changes post-Far-C exposure also incited apoptotic cell death which concomitantly led to significant activation of caspase-3 (p < 0.001). Furthermore, Far-C exhibited its competence in altering the expression of genes involved in apoptosis regulation (Bax, Bad, and Bcl2) along with genes exerting regulatory effects on cell cycle (cyclinD1) and its progression (p21Cip1 and CDK4). The evidence thus clearly shows the preclinical efficacy of Far-C against HepG2 cells. However, further mechanistic investigations deciphering the alteration of different pathways post-Far-C exposure will be highly beneficial.

Plantaricin BM-1 decreases viability of SW480 human colorectal cancer cells by inducing caspase-dependent apoptosis

“…In this study, we focused on the suppression of SW480 cell treated with plantaricin BM-1 via induction of apoptosis. In previous studies, Hoechst33342 staining was used to observe the increase in fluorescence intensity of apoptotic cells, as well as aggregation and fragmentation of the nuclei ( Goud et al, 2020 ; Wu et al, 2020 ; Huang et al, 2021 ; Alafnan et al, 2022 ). This was similar to our results ( Figure 3 ) using Hoechst33342 staining, which demonstrated that after receiving plantaricin BM-1 for 1 h, the initiation of the apoptosis process via enhanced chromatin condensation and the number of cells with condensed nuclei of high fluorescence intensity was increased dose-dependently.…”

High therapeutic efficacy of 5‐Fluorouracil‐loaded exosomes against colon cancer cells

“…The onset of apoptosis within HCT116 cells, upon exposure with 5‐FU‐loaded exosomes, was evaluated using acridine orange and ethidium bromide dual staining (Alafnan et al, 2022). In brief, 5 × 10 5 HCT116 cells were subjected to 5‐FU‐loaded exosomes (equivalent to 2.5 μg and 5 μg 5‐FU) for 24 h. Thereafter, the cells were stained with a mixture of AO/EtBr stain (10 μl each) and were incubated in the dark for 15 min.…”

“…The effect of 5‐FU‐loaded exosomes on ΔΨm in HCT116 cells was estimated using Rh123 (Alafnan et al, 2022). Approximately 2 × 10 5 HCT116 cells were seeded in each well of a 96‐well plate and left overnight in humidified atmosphere at 37°C with 5% CO 2 .…”

“…Caspase‐3 activity in HCT116 cells was quantified colorimetrically following the manufacturer's instructions. Briefly, 3 × 10 6 HCT116 cells were treated with 5‐FU‐loaded exosomes (equivalent to 2.5 μg and 5 μg 5‐FU) for 24 h (Alafnan et al, 2022). Thereafter, the cells were lysed and centrifuged at 8944 g for 1 min at 4°C.…”

“…Briefly, 3 × 10 6 HCT116 cells were treated with 5-FU-loaded exosomes (equivalent to 2.5 μg and 5 μg 5-FU) for 24 h (Alafnan et al, 2022). Thereafter, the cells were lysed and centrifuged at 8944 g for 1 min at 4°C.…”