Endomorphin-2 Analog Inhibits the Growth of DLD-1 and RKO Human Colon Cancer Cells by Inducing Cell Apoptosis.

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.

• Authors: Guanghua Li, Bo Li, Jingang Song, Na Wang, Zhuanglei Gao
• Journal: Medical science monitor : international medical journal of experimental and clinical research (Med Sci Monit) Vol. 26 Pg. e921251 (Apr 27 2020) ISSN: 1643-3750 [Electronic] United States
• PMID: 32336747 (Publication Type: Journal Article)

Chemical References

• Oligopeptides
• Reactive Oxygen Species
• bcl-2-Associated X Protein
• endomorphin 2
• Cytochromes c
• PARP1 protein, human
• Poly (ADP-Ribose) Polymerase-1
• CASP3 protein, human
• CASP9 protein, human
• Caspase 3
• Caspase 9

Topics

• Apoptosis (drug effects)
• Caspase 3 (metabolism)
• Caspase 9 (metabolism)
• Cell Cycle
• Cell Line, Tumor
• Cell Proliferation (drug effects)
• Colon (metabolism)
• Colonic Neoplasms (drug therapy, metabolism)
• Cytochromes c (metabolism)
• Humans
• Oligopeptides (pharmacology)
• Poly (ADP-Ribose) Polymerase-1 (metabolism)
• Reactive Oxygen Species (metabolism)
• bcl-2-Associated X Protein (metabolism)