| Customization: | Available |
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| CAS No.: | 307297-39-8 |
| Formula: | C14h22n4o9 |
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Objective: A type of endocrine gland in animals, the pineal gland, can secrete a peptide called "Epithalamine", which can be artificially synthesized into a tetrapeptide called "Ala Glu Asp Gly". There are reports that Epithalon has a certain tumor inhibitory effect in animals. The purpose of this study is to investigate the effects of Epithalon on the growth, telomere length, and telomerase activity of human liver cancer cells HepG2. At the same time, by comparing the changes in cell growth, telomere length, and telomerase activity of human fetal liver cells L-02 treated with Epithalon, the effects of Epithalon on telomeres and telomerase were studied, and the possible mechanism of action was explored. Method: (1) Epithalon tetrapeptide was prepared by Fmoc solid-phase peptide synthesis method, purified by high-performance liquid chromatography (HPLC), and analyzed and identified by NMR and mass spectrometry. (2) Detect the effects of Epithalon tetrapeptide on the proliferation and viability of human liver cancer cell line HepG2 and human fetal liver cell line L-02 using MTT assay; Observe the growth of cells after medication using cell morphology, and detect the cell cycle using flow cytometry. (3) According to the MTT test results, after a certain concentration of medication for a period of time, telomerase repeat amplification pyrophosphate enzyme-linked luminescence (TRAP-ELIDA) was used to detect telomerase activity in human liver cancer cell line HepG2 and human fetal liver cell line L-02, and telomere length was measured by flow cytometry fluorescence in situ hybridization (FLOW-FISH). Result: (1) The combination of nuclear magnetic resonance and mass spectrometry analysis identified that the synthesized product was indeed Epithalon tetrapeptide. (2) The MTT method was used to detect the effect of Epithalon tetrapeptide on the proliferation and viability of human liver cancer cell line HepG2. The results showed that the low concentration (0.00256~0.0256) μ Mol/L and high concentration (25.6 μ Epithalon with a concentration of mol/L has little effect on cell growth, while 0.256-2.56 μ Epithalon at a concentration of mol/L has a certain inhibitory effect on HepG2 cells. Flow cytometry detection of cell cycle indicates that Epithalon has a certain S-phase blocking effect. The effect of Epithalon on the proliferation and viability of human fetal liver cell line L-02 shows: 2.56 μ Mol/L~25.6 μ Epithalon at a concentration of mol/L has a promoting effect on the proliferation of L-02 cells. (3) TRAP-ELIDA was used to detect telomerase activity in human liver cancer cell line HepG2, and flow cytometry fluorescence in situ hybridization (FLOW-FISH) was used to detect changes in telomere length. It was found that the addition of Epithalon had a certain promoting or prolonging effect on telomerase activity and telomere length in HepG2 cells, but this effect was not significant. For L-02 cells, Epithalon can effectively enhance telomerase activity and increase the length of cell telomeres
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