Anticancer research · 2025
Methionine Deprivation-induced Cancer Cell Death and Methylation Changes in Key Genes and Gene Promoters of Prostate Cancer Cell Lines.
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Ramesh Babu Yatin Srinivash, Venkatachalam Kallidaikurichi V
Аннотация
BACKGROUND/AIM: While normal cells are highly regulated, cancer cells take a dysregulated path which bolsters their survival. Currently, a limited number of uniform treatments are available for cancer cure. Our goal was to deprive cancer cells of the key nutrient methionine and determine what effect it would have on cell death and alterations in DNA methylation of prostate cancer cells.
MATERIALS AND METHODS: PC3 and other cell lines were transfected with plasmid gene constructs for methionine gamma lyase deaminase (MEGL), a methionine-degrading enzyme, targeted for expression in the cytoplasm (cMEGL) or the nucleus (nMEGL). For assessing cell death due to MEGL-mediated methionine deprivation, a standard 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was used. PC3, and DU145 prostate cancer cells were selected for whole-methylome sequencing to determine the effects of MEGL expression. Key gene products comprising the Prolaris Molecular Score, specifically 31 cell-cycle progression genes, were chosen for assessing putative differences in methylome.
RESULTS: Treatment with MEGL gene targeted for expression in either the cytoplasm or nucleus caused significant cell death, similar to that due to the anticancer drug methotrexate. Azacytidine showed no effect on PC3 cell death. Propargylglycine, an inhibitor of MEGL, prevented cell death. Methylome analysis showed increased methylation of two genes: Spindle and kinetochore-associated complex subunit 1 (SKA1), origin recognition complex subunit 6 (ORC6L), and reduced methylation of six promoters: BUB1 mitotic checkpoint serine/threonine kinase B (BUB1B), PDZ binding kinase (PBK), baculoviral IAP repeat-containing 5 (BIRC5), centromere protein M (CENPM), DNA topoisomerase II alpha (TOP2A), minichromosome maintenance 10 replication initiation factor (MCM10), upon forced expression of MEGL.
CONCLUSION: Methionine deprivation through MEGL-targeted gene therapy may be a viable option for inducing cancer cell death compared to unrestricted levels of methionine.
MATERIALS AND METHODS: PC3 and other cell lines were transfected with plasmid gene constructs for methionine gamma lyase deaminase (MEGL), a methionine-degrading enzyme, targeted for expression in the cytoplasm (cMEGL) or the nucleus (nMEGL). For assessing cell death due to MEGL-mediated methionine deprivation, a standard 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was used. PC3, and DU145 prostate cancer cells were selected for whole-methylome sequencing to determine the effects of MEGL expression. Key gene products comprising the Prolaris Molecular Score, specifically 31 cell-cycle progression genes, were chosen for assessing putative differences in methylome.
RESULTS: Treatment with MEGL gene targeted for expression in either the cytoplasm or nucleus caused significant cell death, similar to that due to the anticancer drug methotrexate. Azacytidine showed no effect on PC3 cell death. Propargylglycine, an inhibitor of MEGL, prevented cell death. Methylome analysis showed increased methylation of two genes: Spindle and kinetochore-associated complex subunit 1 (SKA1), origin recognition complex subunit 6 (ORC6L), and reduced methylation of six promoters: BUB1 mitotic checkpoint serine/threonine kinase B (BUB1B), PDZ binding kinase (PBK), baculoviral IAP repeat-containing 5 (BIRC5), centromere protein M (CENPM), DNA topoisomerase II alpha (TOP2A), minichromosome maintenance 10 replication initiation factor (MCM10), upon forced expression of MEGL.
CONCLUSION: Methionine deprivation through MEGL-targeted gene therapy may be a viable option for inducing cancer cell death compared to unrestricted levels of methionine.