Anti-apoptosis Effect of Decoy Receptor 3 in Cholangiocarcinoma Cell Line TFK-1
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Published on: January 11, 2019
Ying-Chen Xu1, Jing Cui2, Li-Jun Zhang1
1Department of Hepatobiliary Surgery, Surgical Laboratory, Beijing Tong Ren Hospital, Capital Medical University, Beijing 100730, China.
Background:
Decoy receptor 3 (DcR3) is a protein with anti-apoptotic effect that belongs to the tumor necrosis factor receptor superfamily. DcR3 is highly expressed in a variety of malignant tumors including cholangiocarcinoma and its expression was found to be related to the clinical stage, the invasion, and the metastasis of the tumor. This in vitro study aimed to investigate the effect of downregulated expression of DcR3 on cell viability, cell apoptosis, and cell cycle in cholangiocarcinoma cell line TFK-1.
Methods:
Three different cell lines were cultured: human cholangiocarcinoma TFK-1, human biliary epithelial carcinoma HuCCT-1, and human cholangiocarcinoma RBE. The cholangiocarcinoma cell line with the highest expression of DcR3 was selected for further investigation. The expression of DcR3 was silenced/knocked down by transfection with DcR3-siRNA in the selected cell line. Various biological phenotype parameters such as cell viability, apoptosis, and cell cycle were observed.
Results:
The mRNA and protein levels of DcR3 were measured in the three cell lines, and TFK-1 was selected. After the treatment with DcR3-siRNA for 48 h, DcR3 mRNA and protein expression in the treatment group were 38.45% (P < 0.01) and 48.03% (P < 0.05) of that of the control, respectively. It was found that the cell viability decreased to 61.87% of the control group (P < 0.01) after the downregulation of DcR3 in cholangiocarcinoma cell line TFK-1 by transfection with DcR3-siRNA, while the percentage of apoptotic cells was 2.98 times as compared with the control group (P < 0.05). Compared with the control group the ratio of G0/G1increased, and the ratio of G2/M decreased in the treatment group. However, the differences were not statistically significant.
Conclusions:
The effect of DcR3 on the growth and apoptosis of cholangiocarcinoma has been demonstrated. DcR3 is not only a predictive marker for malignant tumor but it is also likely to be a potential target for cancer gene therapy. Further studies should focus on exploring the binding ligand of DcR3, the signaling pathway involved, and the molecular mechanism for the regulation of DcR3 expression in cholangiocarcinoma.
Insights
Downregulating decoy receptor 3 (DcR3) in cholangiocarcinoma cells significantly reduced cell viability and increased apoptosis. This suggests DcR3 is a potential therapeutic target for cholangiocarcinoma gene therapy.
Area of Science:
- Oncology
- Molecular Biology
- Cancer Research
Background:
- Decoy receptor 3 (DcR3), a member of the tumor necrosis factor receptor superfamily, exhibits anti-apoptotic properties.
- DcR3 is overexpressed in various cancers, including cholangiocarcinoma, correlating with tumor progression and metastasis.
Purpose of the Study:
- To investigate the in vitro effects of downregulating DcR3 expression on cholangiocarcinoma cell viability, apoptosis, and cell cycle.
- To evaluate DcR3 as a potential therapeutic target in cholangiocarcinoma.
Main Methods:
- Three cholangiocarcinoma cell lines (TFK-1, HuCCT-1, RBE) were cultured and DcR3 expression was analyzed.
- TFK-1 cells, exhibiting the highest DcR3 expression, were selected for gene silencing using DcR3-siRNA.
- Cell viability, apoptosis rates, and cell cycle distribution were assessed post-transfection.
Main Results:
- DcR3 mRNA and protein levels were significantly reduced by DcR3-siRNA in TFK-1 cells.
- Downregulation of DcR3 led to a significant decrease in cholangiocarcinoma cell viability and a significant increase in apoptosis.
- While cell cycle distribution showed trends (increased G0/G1, decreased G2/M), these changes were not statistically significant.
Conclusions:
- DcR3 plays a crucial role in the growth and survival of cholangiocarcinoma cells.
- DcR3 is a potential predictive marker and a promising therapeutic target for cholangiocarcinoma gene therapy.
- Further research is needed to elucidate DcR3's ligand, signaling pathways, and regulatory mechanisms.
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