NaHS inhibits cell proliferation by enhancing oxidative stress and apoptosis in HCT116 cells
Aysegül Öztürk1, Şeyma Taştemur2, Ahmet Ozan Kaleci3
1Department of Therapy and Rehabilitation, Vocational School of Health Services, Sivas Cumhuriyet University, Sivas, Türkiye.
Background:
Colorectal cancer remains a leading cause of cancer-related mortality, and current treatments are often limited by toxicity and therapeutic resistance. This study investigated the antiproliferative effects of the fast-releasing hydrogen sulfide (H₂S) donor sodium hydrosulfide (NaHS) in HCT116 colorectal cancer cells and explored the underlying mechanisms related to oxidative stress, DNA damage, apoptosis, inflammation, and key growth-related signaling pathways.
Materials And Methods:
HCT116 colorectal cancer cells and CCD-18Co normal colon fibroblast cells were cultured under standard conditions and treated with increasing concentrations of sodium hydrosulfide (NaHS). Cell viability was assessed using the XTT assay to determine antiproliferative activity and half-maximal inhibitory concentration (IC₅₀) values. Oxidative stress was evaluated by measuring total antioxidant status (TAS), total oxidant status (TOS), and intracellular reactive oxygen species (ROS) generation. Apoptosis and signaling-related proteins were analyzed by enzyme-linked immunosorbent assay (ELISA)-based quantification of BCL2-associated agonist of cell death (BAD), caspase-3, B-cell lymphoma 2 (BCL-2), phosphoinositide 3-kinase (PI3K), mitogen-activated protein kinase (MAPK), and nuclear factor kappa B (NF-κB), supported by Annexin V flow cytometry. Oxidative DNA damage and apoptotic activation were further evaluated using immunofluorescence staining for 8-hydroxy-2'-deoxyguanosine (8-OHdG) and cleaved caspase-3.
Results:
NaHS significantly reduced HCT116 cell viability in a concentration- and time-dependent manner, with IC₅₀ values of 4.56, 2.549, and 1.688 mM after 24, 48, and 72 h of treatment, respectively. In contrast, lower concentrations of NaHS did not significantly affect the viability of CCD-18Co normal colon fibroblast cells. NaHS treatment induced oxidative stress, as evidenced by decreased TAS, increased TOS, and elevated intracellular ROS levels. Apoptotic responses were associated with increased BAD and caspase-3 levels, decreased BCL-2 expression, and increased early and late apoptotic cell populations. NaHS also increased 8-OHdG and cleaved caspase-3 immunofluorescence signals. In addition, reduced levels of PI3K-, MAPK-, and NF-κB-related proteins were observed following NaHS treatment.
Conclusion:
NaHS exerts potent antiproliferative effects in colorectal cancer cells by inducing oxidative stress-mediated apoptosis and inhibiting key survival and inflammatory signaling pathways.
Insights
Sodium hydrosulfide (NaHS) effectively inhibits colorectal cancer cell growth by inducing oxidative stress and apoptosis. This hydrogen sulfide donor shows promise as a therapeutic agent, targeting key survival pathways in cancer cells.
Area of Science:
- Biochemistry
- Cell Biology
- Oncology
Background:
- Colorectal cancer is a major cause of cancer mortality.
- Current treatments face limitations due to toxicity and resistance.
- Investigating novel therapeutic agents like hydrogen sulfide donors is crucial.
Purpose of the Study:
- To evaluate the antiproliferative effects of sodium hydrosulfide (NaHS) on HCT116 colorectal cancer cells.
- To elucidate the mechanisms underlying NaHS-induced cell death, including oxidative stress, DNA damage, and apoptosis.
- To examine the impact of NaHS on key signaling pathways involved in cancer cell survival and inflammation.
Main Methods:
- HCT116 cells were treated with varying concentrations of NaHS.
- Cell viability was assessed using the XTT assay.
- Oxidative stress markers (TAS, TOS, ROS), apoptosis (BAD, caspase-3, BCL-2, Annexin V), DNA damage (8-OHdG), and signaling proteins (PI3K, MAPK, NF-κB) were quantified.
Main Results:
- NaHS demonstrated significant, dose- and time-dependent antiproliferative effects on HCT116 cells.
- NaHS induced oxidative stress, characterized by altered TAS, TOS, and elevated ROS.
- Apoptosis was triggered, evidenced by changes in apoptotic markers and increased DNA damage, while key survival pathways were inhibited.
Conclusions:
- NaHS exhibits potent antiproliferative activity against colorectal cancer cells.
- The mechanism involves induction of oxidative stress-mediated apoptosis.
- NaHS inhibits critical survival and inflammatory signaling pathways, suggesting therapeutic potential.
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