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Long non-coding RNA GAS5 promotes natural killer cell cytotoxicity against gastric cancer by regulating miR-18a
M F Wei1,2, Z S Gu1, L L Zheng1
1Department of Integrated TCM & Western Medicine, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, China.
Abstract:
Natural killer (NK) cells play significant roles in spontaneous antitumor response in multiple cancers, including gastric cancer. Currently, lncRNAs were identified as essential modulators in the development of NK cells via competing for the target miRNA. However, the regulatory mechanism of GAS5 in NK cells remains largely elusive. The expressions of GAS5 and miR-18a in NK cells were measured by qRT-PCR. The killing effects of NK cells were conducted by lactate dehydrogenase (LDH) assay. Detection of IFN-γ and TNF-α level was carried out using ELISA assay. The interaction between GAS5 and miR-18a was determined by the luciferase reporter system and RIP assay, respectively. We found that GAS5 expression was downregulated while miR-18a expression was upregulated in primary NK cells isolated from GC patient compared with the healthy controls. Moreover, activation of NK cells stimulated by IL-2 enhanced the secretion of IFN-γ, TNF-α, and the expression of GAS5. The deficiency of GAS5 significantly suppressed the secretion of IFN-γ and TNF-α as well as the killing effect of NK cells. Subsequently, luciferase reporter and RIP assay confirmed the interaction between GAS5 and miR-18a. In addition, miR-18a inhibitor attenuated GAS5 silencing induced inhibition of the cytotoxicity of activated NK cells. In conclusion, GAS5 promotes the killing effect of the natural killer cells against GC by regulating miR-18a, providing promising strategies for NK cells based antitumor therapies.
Insights
Gastric cancer (GC) natural killer (NK) cells show reduced GAS5 and increased miR-18a. GAS5 enhances NK cell antitumor activity against GC by regulating miR-18a.
Area of Science:
- Immunology
- Molecular Biology
- Oncology
Background:
- Natural killer (NK) cells are crucial for spontaneous antitumor responses in cancers like gastric cancer (GC).
- Long non-coding RNAs (lncRNAs) modulate NK cell development by interacting with microRNAs (miRNAs).
- The specific role of GAS5 in NK cell function within the context of GC remains unclear.
Purpose of the Study:
- To investigate the regulatory mechanism of GAS5 in NK cells.
- To determine the relationship between GAS5, miR-18a, and NK cell activity in gastric cancer.
- To explore the potential of GAS5 as a therapeutic target for NK cell-based antitumor strategies.
Main Methods:
- Quantitative real-time PCR (qRT-PCR) for GAS5 and miR-18a expression.
- Lactate dehydrogenase (LDH) assay to assess NK cell cytotoxicity.
- Enzyme-linked immunosorbent assay (ELISA) for IFN-γ and TNF-α levels.
- Luciferase reporter and RNA immunoprecipitation (RIP) assays to confirm GAS5-miR-18a interaction.
Main Results:
- GAS5 was downregulated and miR-18a upregulated in NK cells from GC patients versus healthy controls.
- IL-2 stimulation enhanced NK cell secretion of IFN-γ, TNF-α, and GAS5 expression.
- GAS5 deficiency impaired NK cell killing effects and cytokine secretion.
- GAS5 directly interacts with miR-18a, and miR-18a inhibition reversed GAS5 silencing-induced NK cell dysfunction.
Conclusions:
- GAS5 promotes NK cell antitumor activity against gastric cancer by negatively regulating miR-18a.
- This interaction highlights GAS5 as a potential therapeutic target for enhancing NK cell-based immunotherapies for GC.
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