150 kDa glycoprotein isolated from Solanum nigrum Linne stimulates caspase-3 activation and reduces inducible nitric oxide production in HCT-116 cells.
This study was carried out to investigate the apoptotic effects of glycoprotein (SNL glycoprotein, 150-kDa) isolated from Solanum nigrum Linne, which has been used as an antipyretic and anticancer agent in folk medicine. We found that SNL glycoprotein consists of carbohydrate content (69.74%) and protein content (30.26%), which contains more than 50% hydrophobic amino acids such as glycine and proline. SNL glycoprotein showed remarkable cytotoxic and apoptotic effects at 40 microg/ml of SNL glycoprotein for 4 h in HCT-116 cells. In the activity of the apoptotic related proteins [caspase-3 and poly(ADP-ribose)polymerase (PARP)], the results showed that SNL glycoprotein (40 microg/ml) has a stimulatory effect on caspase-3 activation and PARP cleavage in HCT-116 cells. Moreover, SNL glycoprotein blocked nuclear factor-kappa B (NF-kappaB) activation and reduced inducible nitric oxide (iNO) production. Interestingly, pyrrolidine dithiocarbamate (PDTC, for NF-kappaB inhibitor) and N omega-Nitro-L-arginine methylester hydrochloride (L-NAME, for NO inhibitor) effectively stimulated the caspase-3 activation in HCT-116 cells. The results in this experiment indicated that SNL glycoprotein induces apoptosis through the NF-kappaB activation and inducible nitric oxide (iNO) production in HCT-116 cells. Here, we speculate that SNL glycoprotein is one of the chemotherapeutic agents and of the modulators for apoptotic signals in HCT-116 cells.
Lee
Sei-Jung
SJ
# 521, Molecular Biochemistry Laboratory, Institute of Biotechnology, Chonnam National University, Kwangju, 300 Yongbong-Dong 500-757, South Korea.
Lim
Kye-Taek
KT
D017209:Apoptosis / Q000187:drug effects; D053148:Caspase 3 / Q000378:metabolism*; D061945:Caspase Inhibitors; D002470:Cell Survival / Q000187:drug effects; D053938:DNA Fragmentation / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D004789:Enzyme Activation / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006023:Glycoproteins / Q000302:isolation & purification / Q000494:pharmacology*; D045325:HCT116 Cells; D006801:Humans; D007770:L-Lactate Dehydrogenase / Q000032:analysis / Q000378:metabolism; D016328:NF-kappa B / Q000037:antagonists & inhibitors / Q000378:metabolism; D019331:NG-Nitroarginine Methyl Ester / Q000494:pharmacology; D009569:Nitric Oxide / Q000037:antagonists & inhibitors / Q000096:biosynthesis* / Q000378:metabolism; D019001:Nitric Oxide Synthase / Q000037:antagonists & inhibitors / Q000378:metabolism; D009842:Oligopeptides / Q000494:pharmacology; D010940:Plant Proteins / Q000302:isolation & purification / Q000494:pharmacology*; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism; D011759:Pyrrolidines / Q000494:pharmacology; D015398:Signal Transduction / Q000187:drug effects; D013859:Thiocarbamates / Q000494:pharmacology
D061945:Caspase Inhibitors; D004791:Enzyme Inhibitors; D006023:Glycoproteins; D016328:NF-kappa B; D009842:Oligopeptides; D010940:Plant Proteins; D011759:Pyrrolidines; D013859:Thiocarbamates; C099158:acetyl-aspartyl-glutamyl-valyl-aspartal; C508662:glycoprotein 150-kDa, Solanum nigrum; C020972:pyrrolidine dithiocarbamic acid; D009569:Nitric Oxide; D007770:L-Lactate Dehydrogenase; D019001:Nitric Oxide Synthase; D011065:Poly(ADP-ribose) Polymerases; D053148:Caspase 3; D019331:NG-Nitroarginine Methyl Ester