27 April 2020 : Laboratory Research
Endomorphin-2 Analog Inhibits the Growth of DLD-1 and RKO Human Colon Cancer Cells by Inducing Cell Apoptosis
Guanghua Li1BCDE, Bo Li2BCD, Jingang Song3C, Na Wang4BC, Zhuanglei Gao1ACDEF*DOI: 10.12659/MSM.921251
Med Sci Monit 2020; 26:e921251
Abstract
BACKGROUND: In developed countries, colon cancer is a leading cause of cancer-associated mortality. Dietary changes have resulted in an increased incidence of colon cancer in Asia. This study aimed to investigate the effects of the structural analog of endomorphin-2 (H-Tyr-Pro-Phe-Phe-NH₂) on human colon cancer cells in vitro.
MATERIAL AND METHODS: Human DLD-1 and RKO colon cancer cells and CCD-18Co normal human colonic fibroblasts were treated with increasing doses of the structural analog of endomorphin-2. Cells underwent the MTT assay, fluorescence confocal flow cytometry, and Hoechst 33258 staining to investigate cell proliferation, the cell cycle, and apoptosis. Western blot was used to measure the expression levels of poly(ADP-ribose) polymerase-1 (PARP-1), cytochrome c, caspase-3, and caspase-9. The 2’,7’-dichlorofluorescein diacetate (DCFH-DA) fluorescence method measured reactive oxygen species (ROS).
RESULTS: Cell proliferation of DLD-1 and RKO cells was inhibited by the endomorphin-2 analog in a dose-dependent manner, and a 100 µM dose reduced DLD-1 and RKO cell proliferation by 28% and 23%, respectively, at 72 h. Endomorphin-2 analog induced cell apoptosis and the generation of ROS, activated caspase-3 and caspase-9, and increased the levels of p53 and cytochrome c release, and down-regulated of Akt activation in DLD-1 and RKO cells in a dose-dependent manner. Treatment of the DLD-1 and RKO cells with the endomorphin-2 analog increased the expression of Bax and reduced the expression of Bcl-2.
CONCLUSIONS: Endomorphin-2 analog inhibited colon cancer cell proliferation, activated apoptosis, and down-regulated Akt phosphorylation of human DLD-1 and RKO colon cancer cells in vitro in a dose-dependent manner.
Keywords: Chromatin Assembly and Disassembly, Cell Cycle, Colon, Colonic Neoplasms, Cytochromes c, Oligopeptides, bcl-2-Associated X Protein
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