Solidago securrens total saponin combined with cisplatin induce oxidative stress and promote mito-chondrial-dependent apoptosis of liver cancer HepG2 cells
Objective To evaluate the impact of Solidago securrens total saponin(SS),in associa-tion with cisplatin(DDP),on proliferation and apoptosis of human hepatocellular carcinoma HepG2 cells,along with elucidating the potential molecular mechanisms involved.Methods The HepG2 cells were ex-posed to control group(Phosphate Buffered Saline),SS group(320 μg/ml),low-dose DDP group(2μmol/ml),high-dose DDP group(5 μmol/ml)and combination group(SS 320 μg/ml+DDP 2 μmol/ml)for 24 h.Cell viability was measured using MTT assay and colony formation,cell migration was measured using scratch wound healing assay,ROS levels were observed using DCFH-DA probe,mitochondrial mem-brane potential were observed using JC-1 probe,the apoptosis was evaluated by Hoechst 33342 probe,and the protein abundance of Nrf2,heme oxygenase-1(HO-1),B cell lymphoma/leukemia-2 associated X pro-tein(bax),B cell lymphoma/leukemia-2(bcl-2)and cleaved-cysteinyl aspartate-specific protease-3(cleaved-Caspase-3)were measured using Western blotting.Results Compared to the control group,the proliferation of SS group,low-dose DDP group,high-dose DDP group and combination group[(68.75±4.44)％,(82.13±3.19)％,(54.11±4.79)％,(49.92±3.83)％vs.(103.62±2.71)％,F=24.66,P＜0.05]and migration[(28.86±2.35)％,(25.78±3.45)％,(6.85±1.54)％,(5.17±1.98)％vs.(86.85±4.79)％,F=88.56,P＜0.05]capability of HepG2 cells were diminished,SOD[(65.85±6.58)％,(77.86±6.73)％,(28.65±4.43)％,(26.07±4.55)％vs.(103.19±1.73)％,F=40.96,P＜0.05]and GSH levels[(72.29±6.37)％,(77.01±5.14)％,(31.75±6.01)％,(21.73±3.38)％vs.(102.78±3.60)％,F=43.71,P＜0.05]were decreased,the mitochondrial membrane potential were depolarized[(2.76±0.21),(2.33±0.25),(4.91±0.43),(5.85±0.28)vs.(1.03±0.11),F=54.47,P＜0.05],concurrently with a amplified ROS[(3.36±0.44),(2.71±0.37),(6.85±0.62),(7.82±0.51)vs.(1.13±0.16),F=50.83,P＜0.05]and elevated apoptotic rate[(14.35±2.15)％,(14.98±3.77)％,(55.19±3.46)％,(55.82±4.31)％vs.(1.38±0.33)％,F=57.55,P＜0.05]in all groups.In addition,HO-1[(0.34±0.05),(0.46±0.04),(0.16±0.03),(0.11±0.04)vs.(1.09±0.01),F=111.20,P＜0.05]and bcl-2 levels[(0.42±0.04),(0.52±0.03),(0.31±0.02),(0.17±0.03)vs.(1.03±0.05),F=147.90,P＜0.05]were downregulated,Nrf2 nuclear translocation was suppressed[(0.45±0.03),(0.59±0.04),(0.48±0.04),(0.21±0.03)vs.(1.06±0.03),F=66.59,P＜0.05],bax[(3.35±0.53),(2.70±0.45),(6.7±0.39),(7.82±0.33)vs.(1.06±0.03),F=59.95,P＜0.05]and cleaved-Caspase-3 levels[(2.75±0.26),(2.63±0.38),(5.93±0.29),(6.44±0.36)vs.(1.10±0.15),F=62.63,P＜0.05].Conclusion SS combined with DDP can impairs the proliferative potential and migratory capacity of HepG2 cells.The underlying mechanism might involve suppression of Nrf2 pathway activity and upregu-lation of apoptosis-related protein.
Solidago securrens total saponinCisplatinHepatocellular carcinomaApoptosisOxidative stress
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