Induction of ROS-dependent mitochondria-mediated intrinsic apoptosis in MDA-MB-231 cells by glycoprotein from Codium decorticatum.
Marine macroalgae consist of a range of bioactive molecules exhibiting different biological activities, and many of these properties are attributed to sulfated polysaccharides, fucoxanthin, phycobiliproteins, and halogenated compounds. In this study, a glycoprotein (GLP) with a molecular mass of ∼48 kDa was extracted and purified from Codium decorticatum and investigated for its cytotoxic properties against human MDA-MB-231 breast cancer cells. The IC₅₀ values of GLP against MDA-MB-231 and normal breast HBL-100 cells (control) were 75 ± 0.23 μg/mL (IC₂₅), 55 ± 0.32 μg/mL (IC₅₀), and 30 ± 0.43 μg/mL (IC₇₅) and 90 ± 0.57 μg/mL (IC₂₅), 80 ± 0.48 μg/mL (IC₅₀), and 60 ± 0.26 μg/mL (IC₇₅), respectively. Chromatin condensation and poly(ADP-ribose) polymerase (PARP) cleavage studies showed that the GLP inhibited cell viability by inducing apoptosis in MDA-MB-231 cells. Induction of mitochondria-mediated intrinsic apoptotic pathway by GLP was evidenced by the events of loss of mitochondrial membrane potential (ΔΨ(m)), bax/bcl-2 dysregulation, cytochrome c release, and activation of caspases 3 and 9. Apoptosis-associated factors such as reactive oxygen species (ROS) formation and loss of ΔΨ(m) were evaluated by DCFH-DA staining and flow cytometry, respectively. Cell cycle arrest of G₂/M phase and expression of apoptosis associated proteins were determined using flow cytometry and Western blotting, respectively.
Proteomics and Molecular Cell Physiology Laboratory, Department of Zoology, Bharathiar University , Coimbatore, Tamil Nadu, India.
Ramar
R
Thangam
Dharmaraj
D
Senthilkumar
Veeraperumal
V
Suresh
Malairaj
M
Sathuvan
Srinivasan
S
Sivasubramanian
Kalailingam
K
Pazhanichamy
Praveen Kumar
PK
Gorlagunta
Soundarapandian
S
Kannan
Palani
P
Gunasekaran
Ramasamy
R
Rengasamy
Jayanthi
J
Sivaraman
D017209:Apoptosis / Q000187:drug effects*; D059447:Cell Cycle Checkpoints / Q000187:drug effects; D045744:Cell Line, Tumor; D000460:Chlorophyta / Q000737:chemistry*; D006023:Glycoproteins / Q000302:isolation & purification / Q000494:pharmacology*; D006801:Humans; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism*; D017382:Reactive Oxygen Species / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects; D051028:bcl-2-Associated X Protein / Q000235:genetics / Q000378:metabolism
D006023:Glycoproteins; D017382:Reactive Oxygen Species; D051028:bcl-2-Associated X Protein