title: Violacein induces apoptosis in human breast cancer cells through up regulation of BAX, p53 and down regulation of MDM2.
abstract: We aimed to explore the anticancer potential of violacein and its time, dose dependent mechanism of action in human MCF-7 breast cancer cells. We observed, violacein inhibit MCF-7 cells viability in a time and dose-dependent manner, IC50 value was 4.5 μM in 24 h, 1.7 μM in 48 h and 0.51 μM in 72 h. Violacein triggered generation of intra cellular ROS even from the lower doses, significant ROS production was observed from 0.25, 0.45 μM dose range and it is relative to higher doses. Further we fixed 0.45 μM and 4.5 μM as an experimental dose for relative dose dependent analysis. In nuclear staining, after 48 h 0.45 μM dose showed characteristic apoptotic morphological changes such as, 59% of cells in apoptosis and 11% of cells in necrotic stage, also in 72 h we found 68% in apoptosis and 12% in necrotic stage. However, 4.5 μM (IC50) dose of violacein, 78% of cells became apoptotic and 21% in necrotic after 48 h; but in 72 h only 61% cells are in apoptotic, necrosis was increased to 38%. Violacein increased both mitochondrial and extra mitochondrial apoptotic pathway related gene expressions; it was confirmed by increased CYP1A, GPX, GSK3β and TNF-α gene. Further, 0.45 and 4.5 μM of violacein increased apoptotic genes, such as Bax, p53, caspase 3, Fas, FADD and markedly reduced Bcl-2 and MDM2 expression levels to two fold when compared to control. In addition violacein upregulated poly ADP-ribose polymerase (PARP), CDKN1A and caspase-9 significantly (p≤0.05) when compared to control. Relative quantification of caspase-8 was differently expressed; there were no changes in 0.45 μM, but in 4.5 μM we found two fold increased caspase-8 expression. In conclusion, lower dose of violacein treatment induced apoptosis in human breast cancer MCF-7 cells through TNF-α and p53 dependent mitochondrial pathways.
authors.affiliation: Cancer Molecular Biology Research Lab, Department of Food and Nutrition, College of Food and Agricultural Sciences, King Saud University, Riyadh 11451, Saudi Arabia. Electronic address: alshatwi@ksu.edu.sa.
authors.forename: Ali A
authors.initials: AA
authors.lastname: Alshatwi
authors.affiliation: Cancer Molecular Biology Research Lab, Department of Food and Nutrition, College of Food and Agricultural Sciences, King Saud University, Riyadh 11451, Saudi Arabia. Electronic address: subashbabu80@gmail.com.
authors.forename: P
authors.initials: P
authors.lastname: Subash-Babu
authors.affiliation: Department of Veterinary Physiology, College of Veterinary Medicine, Chonbuk National University, Biosafety Research Institute, 664-14 1GA, Duckjin-Dong, Duckjin-Gu, Jeonju City, Jeollabuk-Do 561-756, South Korea.
authors.forename: P
authors.initials: P
authors.lastname: Antonisamy
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects* / Q000502:physiology; D001943:Breast Neoplasms / Q000378:metabolism / Q000473:pathology*; D002470:Cell Survival / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D005260:Female; D006801:Humans; D020287:In Situ Nick-End Labeling; D007211:Indoles / Q000494:pharmacology*; D061986:MCF-7 Cells; D051736:Proto-Oncogene Proteins c-mdm2 / Q000096:biosynthesis / Q000187:drug effects; D017382:Reactive Oxygen Species / Q000378:metabolism; D016159:Tumor Suppressor Protein p53 / Q000096:biosynthesis / Q000187:drug effects; D051028:bcl-2-Associated X Protein / Q000096:biosynthesis / Q000187:drug effects
chemical_list: D000970:Antineoplastic Agents; C494826:BAX protein, human; D007211:Indoles; D017382:Reactive Oxygen Species; D016159:Tumor Suppressor Protein p53; D051028:bcl-2-Associated X Protein; C497206:MDM2 protein, human; D051736:Proto-Oncogene Proteins c-mdm2; C063155:violacein
keywords: Apoptosis; MCF-7 cells; MDM2; Violacein; p53