Objective: To explore cytotoxicity of Synsepalum dulcificum(S. dulcificum) Daniell(Sapotaceae) on human colon cancer(HCT-116 and HT-29), human monocytic leukemia(THP-1) and normal(HDFn) cell lines, and its effect on t...Objective: To explore cytotoxicity of Synsepalum dulcificum(S. dulcificum) Daniell(Sapotaceae) on human colon cancer(HCT-116 and HT-29), human monocytic leukemia(THP-1) and normal(HDFn) cell lines, and its effect on the expression of early apoptotic genes, c-fos and c-jun. Methods: Leaf, stem and berry of S. dulcificum were separately extracted by using 2 solvents, 10% ethanol(EtOH) and 80% methanol(MeOH). PrestoB lue~? cell viability assay and q RT-PCR assay were conducted to examine the above objectives respectively. Results: Stem MeOH, stem EtOH, and berry EtOH extracts of S. dulcificum were cytotoxic to HCT-116 and HT-29 human colon cancer cells. For HCT-116, IC_(50) values of these 3 extracts were not significantly different(P>0.05) from that of the positive control bleomycin(IC_(50) of 33.57 μg/mL), while for HT-29, IC_(50) values of these 3 extracts were significantly lower(P<0.05) than that of bleomycin(IC_(50) of 25.24 μg/mL). None of the extracts were cytotoxic to the THP-1 monocytic leukemia cells and HDFn normal human dermal fibroblasts. For both HCT-116 and HT-29, these extracts significantly up-regulated(P<0.05) the expression of c-fos and c-jun compared to the untreated negative control. Conclusions: The results of this study suggest that cytotoxicity of stem MeOH, stem EtOH, and berry EtOH extracts of S. dulcificum on HCT-116 and HT-29 colon cancer cells is due to the induced apoptosis which is caused by the up-regulation of the expression of early apoptotic genes, c-fos and c-jun.展开更多
Objective:To evaluate antioxidant,cytotoxic,and anti-venom capacity of crude bark extracts of Alstonia parvifolia Merr.Methods:Gas chromatography-mass spectrometry(GC-MS)and energy dispersive X-ray analyses were accom...Objective:To evaluate antioxidant,cytotoxic,and anti-venom capacity of crude bark extracts of Alstonia parvifolia Merr.Methods:Gas chromatography-mass spectrometry(GC-MS)and energy dispersive X-ray analyses were accomplished to characterize the chemical constituents of Alstonia parvifolia.Biochemical characterization was evaluated using an inhibitory phospholipase A_(2)(PLA_(2))assay,DPPH,and cytotoxicity assays.Using the constituents listed in the GC-MS analyses,molecular docking was conducted to inspect the binding energies between the chosen compounds and selected PLA_(2) isoforms.Results:GC-MS analyses showed that the Alstonia parvifolia crude extract consisted predominantly of acetylmarinobufogenin(14.89%),γ-sitosterol(10.44%),3-O-methyl-D-glucose(5.88%),3,5-dimethoxy-4-hydroxyphenylacetic acid(5.30%),(2α,5α)-17-methoxyaspidofractinin-3-one(AFM)(4.08%),and 2,3,5,6,7,8,9-heptahydro-1-phenyl-5-(p-chlorophenylimino)-1H-benzo[e][1,4]thiazepine(HPT)(1.37%).The principal elemental components of Alstonia parvifolia were Ca(4.012%)and K(1.496%),as exhibited by energy dispersive X-ray examination.Alstonia parvifolia showed significant free radical scavenging ability(IC_(50):0.287 mg/mL)and was non-cytotoxic to normal HDFn cells(IC_(50)>100μg/mL).Moreover,Alstonia parvifolia was favorably cytotoxic to MCF-7(IC_(50):4.42µg/mL),followed by H69PR,HT-29,and THP-1,with IC_(50) values of 4.94,5.07,and 6.27µg/mL,respectively.Alstonia parvifolia also displayed notable inhibition against PLA_(2) activity of Naja philippinensis Taylor venom with IC_(50) of(15.2±1.8)μg/mL.Docking and cluster analyses projected negative binding energies from AFM(−6.36 to−9.68 kcal/mol),HPT(−7.38 to−9.77 kcal/mol),and acetylmarinobufogenin(−7.22 to−9.59 kcal/mol).These calculations were for the particular interactions of Alstonia parvifolia constituents to PLA_(2) homologues where the utmost affinity was detected in HPT owing to the dipole interactions with amino acid residues.Conclusions:The bark extract of Alstonia parvifolia shows great potential as an anti-venom agent due to its low cytotoxic profile,remarkable PLA_(2) inhibition,and docking binding energies between its bioactive constituents and PLA_(2) homologues.展开更多
文摘Objective: To explore cytotoxicity of Synsepalum dulcificum(S. dulcificum) Daniell(Sapotaceae) on human colon cancer(HCT-116 and HT-29), human monocytic leukemia(THP-1) and normal(HDFn) cell lines, and its effect on the expression of early apoptotic genes, c-fos and c-jun. Methods: Leaf, stem and berry of S. dulcificum were separately extracted by using 2 solvents, 10% ethanol(EtOH) and 80% methanol(MeOH). PrestoB lue~? cell viability assay and q RT-PCR assay were conducted to examine the above objectives respectively. Results: Stem MeOH, stem EtOH, and berry EtOH extracts of S. dulcificum were cytotoxic to HCT-116 and HT-29 human colon cancer cells. For HCT-116, IC_(50) values of these 3 extracts were not significantly different(P>0.05) from that of the positive control bleomycin(IC_(50) of 33.57 μg/mL), while for HT-29, IC_(50) values of these 3 extracts were significantly lower(P<0.05) than that of bleomycin(IC_(50) of 25.24 μg/mL). None of the extracts were cytotoxic to the THP-1 monocytic leukemia cells and HDFn normal human dermal fibroblasts. For both HCT-116 and HT-29, these extracts significantly up-regulated(P<0.05) the expression of c-fos and c-jun compared to the untreated negative control. Conclusions: The results of this study suggest that cytotoxicity of stem MeOH, stem EtOH, and berry EtOH extracts of S. dulcificum on HCT-116 and HT-29 colon cancer cells is due to the induced apoptosis which is caused by the up-regulation of the expression of early apoptotic genes, c-fos and c-jun.
基金supported by the De La Salle University Science Foundation in coordination with the University Research Coordination Office(Project number:18FU2TAY16-3TAY17).
文摘Objective:To evaluate antioxidant,cytotoxic,and anti-venom capacity of crude bark extracts of Alstonia parvifolia Merr.Methods:Gas chromatography-mass spectrometry(GC-MS)and energy dispersive X-ray analyses were accomplished to characterize the chemical constituents of Alstonia parvifolia.Biochemical characterization was evaluated using an inhibitory phospholipase A_(2)(PLA_(2))assay,DPPH,and cytotoxicity assays.Using the constituents listed in the GC-MS analyses,molecular docking was conducted to inspect the binding energies between the chosen compounds and selected PLA_(2) isoforms.Results:GC-MS analyses showed that the Alstonia parvifolia crude extract consisted predominantly of acetylmarinobufogenin(14.89%),γ-sitosterol(10.44%),3-O-methyl-D-glucose(5.88%),3,5-dimethoxy-4-hydroxyphenylacetic acid(5.30%),(2α,5α)-17-methoxyaspidofractinin-3-one(AFM)(4.08%),and 2,3,5,6,7,8,9-heptahydro-1-phenyl-5-(p-chlorophenylimino)-1H-benzo[e][1,4]thiazepine(HPT)(1.37%).The principal elemental components of Alstonia parvifolia were Ca(4.012%)and K(1.496%),as exhibited by energy dispersive X-ray examination.Alstonia parvifolia showed significant free radical scavenging ability(IC_(50):0.287 mg/mL)and was non-cytotoxic to normal HDFn cells(IC_(50)>100μg/mL).Moreover,Alstonia parvifolia was favorably cytotoxic to MCF-7(IC_(50):4.42µg/mL),followed by H69PR,HT-29,and THP-1,with IC_(50) values of 4.94,5.07,and 6.27µg/mL,respectively.Alstonia parvifolia also displayed notable inhibition against PLA_(2) activity of Naja philippinensis Taylor venom with IC_(50) of(15.2±1.8)μg/mL.Docking and cluster analyses projected negative binding energies from AFM(−6.36 to−9.68 kcal/mol),HPT(−7.38 to−9.77 kcal/mol),and acetylmarinobufogenin(−7.22 to−9.59 kcal/mol).These calculations were for the particular interactions of Alstonia parvifolia constituents to PLA_(2) homologues where the utmost affinity was detected in HPT owing to the dipole interactions with amino acid residues.Conclusions:The bark extract of Alstonia parvifolia shows great potential as an anti-venom agent due to its low cytotoxic profile,remarkable PLA_(2) inhibition,and docking binding energies between its bioactive constituents and PLA_(2) homologues.
