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MSC(TRAIL)-mediated HepG2 cell death in direct and indirect co-cultures
Xu-Yong Sun1, Jiang Nong, Ke Qin
1Institute of Transplant Medicine, 303 Hospital of PLA, Nanning, PR China.
Background:
Mesenchymal stem cells (MSCs) have attracted great interest in cancer therapy since the discovery of their tumor tropism. This study was performed to investigate the effects of TNF-related apoptosis-inducing ligand (TRAIL)-engineered MSCs on hepatocellular carcinoma (HCC) cells (HepG2) under different culture conditions.
Materials And Methods:
MSCs engineered with non-secreting TRAIL (MSC(TRAIL-GFP)) (GFP, green fluorescence protein) and secreting TRAIL (MSC(stTRAIL)) were used for the direct co-cultures, and conditioned media (CM) from corresponding cultures were applied to HepG2 as indirect co-cultures. Immunoblotting, ELISA and FACS analysis were used to detect the expression of TRAIL and TRAIL receptors. Cell death was assessed using live/dead assay.
Results:
Death receptor (DR) 5 was identified on the HepG2 cells. The expression of TRAIL was confirmed in the cell lysates (MSC(TRAIL-GFP) >MSC(stTRAIL)) and the conditioned media (MSC(stTRAIL) >MSC(TRAIL-GFP)). Higher cell death was observed in high MSC/HepG2 ratio co-cultures. HepG2 cell death was proportionally related to CM from MSC(TRAIL-GFP) and MSC(stTRAIL).
Conclusion:
MSCs exhibit intrinsic inhibition of HepG2 which is potentiated by TRAIL-transfection.
Insights
Mesenchymal stem cells (MSCs) engineered with TNF-related apoptosis-inducing ligand (TRAIL) show potential in treating hepatocellular carcinoma (HCC). TRAIL-engineered MSCs enhance HepG2 cell death, indicating a promising therapeutic strategy.
Area of Science:
- Oncology
- Cell Biology
- Biotechnology
Background:
- Mesenchymal stem cells (MSCs) possess tumor-homing abilities, making them attractive for cancer therapy.
- Hepatocellular carcinoma (HCC) is a major global health concern.
- Engineering MSCs with therapeutic payloads is a developing strategy in cancer treatment.
Purpose of the Study:
- To investigate the efficacy of TNF-related apoptosis-inducing ligand (TRAIL)-engineered MSCs against hepatocellular carcinoma (HCC) cells.
- To evaluate the impact of different culture conditions on TRAIL-engineered MSCs' anti-cancer effects.
- To assess the role of TRAIL expression levels and secretion in MSC-mediated HepG2 cell death.
Main Methods:
- Co-culture of HepG2 cells with MSCs engineered to express non-secreting (MSC(TRAIL-GFP)) or secreting (MSC(stTRAIL)) TRAIL.
- Indirect co-culture using conditioned media (CM) from MSC cultures.
- Analysis of TRAIL and TRAIL receptor expression (DR5) via immunoblotting, ELISA, and FACS.
- Assessment of HepG2 cell viability using live/dead assays.
Main Results:
- Death receptor 5 (DR5) was detected on HepG2 cells.
- TRAIL expression was higher in MSC(TRAIL-GFP) cell lysates than MSC(stTRAIL).
- TRAIL was secreted more effectively by MSC(stTRAIL) than MSC(TRAIL-GFP).
- Increased HepG2 cell death was observed with higher MSC/HepG2 ratios and with conditioned media from both engineered MSC types.
Conclusions:
- Mesenchymal stem cells possess inherent inhibitory effects on HepG2 cells.
- TRAIL-transfection significantly potentiates the anti-cancer activity of MSCs against HCC.
- TRAIL-engineered MSCs represent a promising approach for hepatocellular carcinoma therapy.
