TRAIL and Taurolidine induce apoptosis and decrease proliferation in human fibrosarcoma
Adrien Daigeler1, Christina Brenzel, Daniel Bulut
1Department of Plastic Surgery, Burn Center, Hand Center, Sarcoma Reference Center, BG-University Hospital Bergmannsheil, Bürkle-de-la-Camp-Platz 1, 44789 Bochum, Germany. adrien.daigeler@rub.de
Background:
Disseminated soft tissue sarcoma still represents a therapeutic dilemma because effective cytostatics are missing. Therefore we tested TRAIL and Tarolidine (TRD), two substances with apoptogenic properties on human fibrosarcoma (HT1080).
Methods:
Viability, apoptosis and necrosis were visualized by TUNEL-Assay and quantitated by FACS analysis (Propidiumiodide/AnnexinV staining). Gene expression was analysed by RNA-Microarray and the results validated for selected genes by rtPCR. Protein level changes were documented by Western Blot analysis. NFKB activity was analysed by ELISA and proliferation assays (BrdU) were performed.
Results And Discussion:
The single substances TRAIL and TRD induced apoptotic cell death and decreased proliferation in HT1080 cells significantly. Gene expression of several genes related to apoptotic pathways (TRAIL: ARHGDIA, NFKBIA, TNFAIP3; TRD: HSPA1A/B, NFKBIA, GADD45A, SGK, JUN, MAP3K14) was changed. The combination of TRD and TRAIL significantly increased apoptotic cell death compared to the single substances and lead to expression changes in a variety of genes (HSPA1A/B, NFKBIA, PPP1R15A, GADD45A, AXL, SGK, DUSP1, JUN, IRF1, MYC, BAG5, BIRC3). NFKB activity assay revealed an antipodal regulation of the several subunits of NFKB by TRD and TRD+TRAIL compared to TRAIL alone.
Conclusion:
TRD and TRAIL are effective to induce apoptosis and decrease proliferation in human fibrosarcoma. A variety of genes seems to be involved, pointing to the NFKB pathway as key regulator in TRD/TRAIL-mediated apoptosis.
Insights
This study shows that TRAIL and Tarolidine (TRD) effectively induce apoptosis and reduce proliferation in human fibrosarcoma cells. The combination therapy demonstrated enhanced efficacy, highlighting the NFKB pathway
Area of Science:
- Oncology
- Molecular Biology
- Pharmacology
Background:
- Disseminated soft tissue sarcoma presents a significant therapeutic challenge due to a lack of effective chemotherapeutics.
- Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and Tarolidine (TRD) possess apoptogenic properties.
- Human fibrosarcoma (HT1080) cells were utilized to investigate the efficacy of TRAIL and TRD.
Purpose of the Study:
- To evaluate the apoptogenic and anti-proliferative effects of TRAIL and TRD on human fibrosarcoma cells.
- To investigate the molecular mechanisms underlying the action of TRAIL and TRD, including gene expression and signaling pathway modulation.
- To assess the synergistic effects of combining TRAIL and TRD in fibrosarcoma treatment.
Main Methods:
- Cell viability, apoptosis, and necrosis were assessed using TUNEL assays and flow cytometry (Propidium Iodide/Annexin V staining).
- Gene expression profiling was performed using RNA-microarray, with validation of key genes via RT-PCR.
- Protein level alterations were analyzed by Western Blot, and NFKB activity was measured using ELISA. Proliferation assays (BrdU) were also conducted.
Main Results:
- Both TRAIL and TRD significantly induced apoptosis and decreased proliferation in HT1080 cells.
- Treatment with TRAIL and TRD altered the expression of multiple genes involved in apoptotic pathways.
- The combination of TRD and TRAIL demonstrated a significant increase in apoptosis compared to single-agent treatments, with notable changes in gene expression profiles.
- NFKB activity was differentially regulated by TRD and the combination therapy compared to TRAIL alone.
Conclusions:
- TRAIL and TRD are effective agents for inducing apoptosis and reducing proliferation in human fibrosarcoma.
- The study implicates the NFKB pathway as a critical regulator in TRAIL/TRD-mediated apoptosis.
- Combined TRD and TRAIL therapy shows promise for treating human fibrosarcoma by enhancing apoptosis and modulating key molecular pathways.
