Cell Death Mechanisms Induced by CLytA-DAAO Chimeric Enzyme in Human Tumor Cell Lines

• Published in: International journal of molecular sciences 2020-11, Vol.21 (22), p.8522
• Main Authors: Fuentes-Baile, María, García-Morales, Pilar, Pérez-Valenciano, Elizabeth, Ventero, María P, Sanz, Jesús M, de Juan Romero, Camino, Barberá, Víctor M, Alenda, Cristina, Saceda, Miguel
• Format: Article
• Language: English
• Subjects: Alanine; Amidase; Amino acid oxidase; Antineoplastic Agents - pharmacology; Apoptosis; Apoptosis Inducing Factor - metabolism; Biopsy; Calcium (intracellular); Calcium (mitochondrial); Calcium - metabolism; Calcium mobilization; Cancer therapies; Caspase; Caspase inhibitors; Cell Death; Cell Line, Tumor; Choline; Colon; Colorectal cancer; Colorectal carcinoma; Colorectal Neoplasms - metabolism; D-Alanine; D-Amino-acid oxidase; D-Amino-Acid Oxidase - metabolism; Deoxyribonucleic acid; Depolarization; Detoxification; DNA; DNA damage; Drug resistance; Enzymes; Gene Expression Profiling; Gene Expression Regulation, Neoplastic; Glioblastoma; HT29 Cells; Humans; Inflammation; L-Alanine; Membrane potential; Membrane Potential, Mitochondrial; Membranes; Mitochondria; N-Acetylmuramoyl-L-alanine Amidase - metabolism; Nanoparticles; Necrosis; NF-kappa B p50 Subunit - metabolism; Oxidative Stress; Pancreas; Pancreatic cancer; Pancreatic Neoplasms - metabolism; Permeability; Poly (ADP-Ribose) Polymerase-1 - metabolism; Poly(ADP-ribose) polymerase; Reactive Oxygen Species - metabolism; RNA, Small Interfering - metabolism; Streptococcus infections; Tumor cell lines
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms21228522

The combination of the choline binding domain of the amidase N-acetylmuramoyl-L-alanine (CLytA)-D-amino acid oxidase (DAAO) (CLytA-DAAO) and D-Alanine induces cell death in several pancreatic and colorectal carcinoma and glioblastoma cell lines. In glioblastoma cell lines, CLytA-DAAO-induced cell death was inhibited by a pan-caspase inhibitor, suggesting a classical apoptotic cell death. Meanwhile, the cell death induced in pancreatic and colon carcinoma cell lines is some type of programmed necrosis. In this article, we studied the mechanisms that trigger CLytA-DAAO-induced cell death in pancreatic and colorectal carcinoma and glioblastoma cell lines and we acquire a further insight into the necrotic cell death induced in pancreatic and colorectal carcinoma cell lines. We have analyzed the intracellular calcium mobilization, mitochondrial membrane potential, PARP-1 participation and AIF translocation. Although the mitochondrial membrane depolarization plays a crucial role, our results suggest that CLytA-DAAO-induced cell death is context dependent. We have previously detected pancreatic and colorectal carcinoma cell lines (Hs766T and HT-29, respectively) that were resistant to CLytA-DAAO-induced cell death. In this study, we have examined the putative mechanism underlying the resistance in these cell lines, evaluating both detoxification mechanisms and the inflammatory and survival responses. Overall, our results provide a better understanding on the cell death mechanism induced by CLytA-DAAO, a promising therapy against cancer.