75 research outputs found
Growth inhibition and apoptosis induced by 2 phenoxymethyl-3H-quinazolin-4-one in HL-60 leukemia cells
Aim: The aim of the study was to investigate anticancer activity of newly synthesized 2-phenoxymethyl-3H-quinazolin-4-one (PMQ). Materials and Methods: Anticancer activity of PMQ was studied towards human HL-60 leukemia cells. Antiproliferative activity of PMQ was determined by direct counting of cells using trypan blue staining technique. Apoptosis and cell cycle profile changes were analysed using internucleosomal DNA fragmentation assay and flow cytometry. Activation of caspases and changes in glutathione level were monitored using colorimetric or luminiscent methods. Results: PMQ induced concentration-dependent cytotoxicity in leukemia cells, with IC50 of 10.8 Β± 0.9 Β΅M. DNA flow cytometry analysis and DNA ladder formation assay indicated that PMQ actively induced apoptosis of cells accompanied by a block of cells in G2/M phase and a marked loss of cells in G0/G1 and S phases. Additionally, the activities of caspase-3 and caspase-9 were increased significantly and a markedly increased level of oxidized glutahione was observed. Inhibition of glutahione synthesis using buthionine sulfoximine sensitized leukemia cells to PMQ, confirming the involvement of ROS in PMQ-induced apoptosis. Conclusion: The results of this study clearly demonstrate that PMQ is a promising anticancer drug showing cytostatic and apoptotic effects toward HL-60 leukemia cells mainly through mitochondrial/caspase-9 dependent pathway.Π¦Π΅Π»Ρ: ΠΈΠ·ΡΡΠΈΡΡ Π°Π½ΡΠΈΠΏΡΠΎΠ»ΠΈΡΠ΅ΡΠ°ΡΠΈΠ²Π½ΡΡ Π°ΠΊΡΠΈΠ²Π½ΠΎΡΡΡ Π½ΠΎΠ²ΠΎΠ³ΠΎ ΡΠΈΠ½ΡΠ΅Π·ΠΈΡΠΎΠ²Π°Π½Π½ΠΎΠ³ΠΎ 2-ΡΠ΅Π½ΠΎΠΊΡΠΈΠΌΠ΅ΡΠΈΠ»-3Π-Ρ
ΠΈΠ½Π°Π·ΠΎΠ»ΠΈΠ½-4-ΠΎΠ½Π° (PMQ).
ΠΠ°ΡΠ΅ΡΠΈΠ°Π»Ρ ΠΈ ΠΌΠ΅ΡΠΎΠ΄Ρ: Π°Π½ΡΠΈΠΏΡΠΎΠ»ΠΈΡΠ΅ΡΠ°ΡΠΈΠ²Π½ΡΡ Π°ΠΊΡΠΈΠ²Π½ΠΎΡΡΡ PMQ ΠΎΠΏΡΠ΅Π΄Π΅Π»ΡΠ»ΠΈ ΠΏΠΎ ΠΎΡΠ½ΠΎΡΠ΅Π½ΠΈΡ ΠΊ ΠΊΠ»Π΅ΡΠΊΠ°ΠΌ Π»Π΅ΠΉΠΊΠΎΠ·Π° Π»ΠΈΠ½ΠΈΠΈ HL-60
Π² ΡΠ΅ΡΡΠ΅ Ρ ΡΡΠΈΠΏΠ°Π½ΠΎΠ²ΡΠΌ ΡΠΈΠ½ΠΈΠΌ ΠΏΡΠΈ ΡΡΠ°Π½Π΄Π°ΡΡΠ½ΠΎΠΌ ΠΏΠΎΠ΄ΡΡΠ΅ΡΠ΅ ΠΊΠ»Π΅ΡΠΎΠΊ. ΠΠΏΠΎΠΏΡΠΎΠ· ΠΈ ΠΊΠ»Π΅ΡΠΎΡΠ½ΡΠΉ ΡΠΈΠΊΠ» ΠΎΡΠ΅Π½ΠΈΠ²Π°Π»ΠΈ Ρ ΠΏΠΎΠΌΠΎΡΡΡ ΠΏΡΠΎΡΠΎΡΠ½ΠΎΠΉ
ΡΠΈΡΠΎΠΌΠ΅ΡΡΠΈΠΈ ΠΈ Π°Π½Π°Π»ΠΈΠ·Π° ΡΡΠ°Π³ΠΌΠ΅Π½ΡΠ°ΡΠΈΠΈ Π²Π½ΡΡΡΠΈΡΠ΄Π΅ΡΠ½ΠΎΠΉ ΠΠΠ. ΠΠΊΡΠΈΠ²Π°ΡΠΈΡ ΠΊΠ°ΡΠΏΠ°Π· ΠΈ ΠΈΠ·ΠΌΠ΅Π½Π΅Π½ΠΈΡ ΡΡΠΎΠ²Π½Ρ Π³Π»ΡΡΠ°ΡΠΈΠΎΠ½Π° ΠΎΠΏΡΠ΅Π΄Π΅Π»ΡΠ»ΠΈ
ΠΊΠΎΠ»ΠΎΡΠΈΠΌΠ΅ΡΡΠΈΡΠ΅ΡΠΊΠΈΠΌΠΈ ΠΈΠ»ΠΈ Π»ΡΠΌΠΈΠ½ΠΈΡΡΠ΅Π½ΡΠ½ΡΠΌΠΈ ΠΌΠ΅ΡΠΎΠ΄Π°ΠΌΠΈ. Π Π΅Π·ΡΠ»ΡΡΠ°ΡΡ: PMQ ΠΈΠ½Π΄ΡΡΠΈΡΡΠ΅Ρ Π΄ΠΎΠ·ΠΎΠ·Π°Π²ΠΈΡΠΈΠΌΡΡ ΡΠΈΡΠΎΡΠΎΠΊΡΠΈΡΠ½ΠΎΡΡΡ
Π² ΠΊΠ»Π΅ΡΠΊΠ°Ρ
Π»ΠΈΠ½ΠΈΠΈ HL-60 (IC50 ΠΏΡΠΈ 10,8 Β± 0,9 ΞΌM). ΠΡΠΈ ΠΏΡΠΎΠ²Π΅Π΄Π΅Π½ΠΈΠΈ Π°Π½Π°Π»ΠΈΠ·Π° ΠΠΠ Ρ ΠΏΡΠΈΠΌΠ΅Π½Π΅Π½ΠΈΠ΅ΠΌ ΠΏΡΠΎΡΠΎΡΠ½ΠΎΠΉ ΡΠΈΡΠΎΠΌΠ΅ΡΡΠΈΠΈ
ΠΈ ΠΎΠΏΡΠ΅Π΄Π΅Π»Π΅Π½ΠΈΠ΅ΠΌ ΡΠΎΡΠΌΠΈΡΠΎΠ²Π°Π½ΠΈΡ Π°ΠΏΠΎΠΏΡΠΈΡΠ΅ΡΠΊΠΎΠΉ Π»Π΅ΡΡΠ½ΠΈΡΡ Π±ΡΠ»ΠΎ ΠΏΠΎΠΊΠ°Π·Π°Π½ΠΎ, ΡΡΠΎ PMQ Π°ΠΊΡΠΈΠ²Π½ΠΎ ΠΈΠ½Π΄ΡΡΠΈΡΡΠ΅Ρ Π°ΠΏΠΎΠΏΡΠΎΠ· ΠΈ Π±Π»ΠΎΠΊΠ°Π΄Ρ
ΠΊΠ»Π΅ΡΠΎΡΠ½ΠΎΠ³ΠΎ ΡΠΈΠΊΠ»Π° Π² G2
/M ΡΠ°Π·Π΅ ΠΌΠΈΡΠΎΠ·Π° ΠΈ Π²ΡΡΠ°ΠΆΠ΅Π½Π½ΠΎΠΉ ΠΏΠΎΡΠ΅ΡΠ΅ΠΉ ΠΊΠ»Π΅ΡΠΎΠΊ Π² G0
/G1
ΠΈ S ΡΠ°Π·Π°Ρ
. ΠΡΠΎΠΌΠ΅ ΡΠΎΠ³ΠΎ, Π±ΡΠ»Π° Π΄ΠΎΡΡΠΎΠ²Π΅ΡΠ½ΠΎ ΠΏΠΎΠ²ΡΡΠ΅Π½Π°
Π°ΠΊΡΠΈΠ²Π½ΠΎΡΡΡ ΠΊΠ°ΡΠΏΠ°Π·Ρ-3 ΠΈ -9 ΠΈ Π²ΡΡΠ°ΠΆΠ΅Π½Π½ΠΎ ΡΠ²Π΅Π»ΠΈΡΠ΅Π½ ΡΡΠΎΠ²Π΅Π½Ρ ΠΎΠΊΠΈΡΠ»Π΅Π½Π½ΠΎΠ³ΠΎ Π³Π»ΡΡΠ°ΡΠΈΠΎΠ½Π°. ΠΡΠΈΠΌΠ΅Π½Π΅Π½ΠΈΠ΅ Π±ΡΡΠΈΠΎΠ½ΠΈΠ½ ΡΡΠ»ΡΡΠΎΠΊΡΠΈΠΌΠΈΠ½Π°
ΠΏΡΠΈΠ²Π΅Π»ΠΎ ΠΊ ΡΠ³Π½Π΅ΡΠ΅Π½ΠΈΡ ΡΠΈΠ½ΡΠ΅Π·Π° Π³Π»ΡΡΠ°ΡΠΈΠΎΠ½Π° ΠΈ ΠΏΠΎΠ²ΡΡΠ΅Π½ΠΈΡ ΡΡΠ²ΡΡΠ²ΠΈΡΠ΅Π»ΡΠ½ΠΎΡΡΠΈ ΠΊΠ»Π΅ΡΠΎΠΊ HL-60 ΠΊ PMQ, ΡΡΠΎ ΠΏΠΎΠ΄ΡΠ²Π΅ΡΠΆΠ΄Π°Π΅Ρ ΡΠ°ΠΊΡ
ΡΡΠ°ΡΡΠΈΡ Π Π€Π Π² PMQ-ΠΈΠ½Π΄ΡΡΠΈΡΠΎΠ²Π°Π½Π½ΠΎΠΌ Π°ΠΏΠΎΠΏΡΠΎΠ·Π΅. ΠΡΠ²ΠΎΠ΄Ρ: PMQ ΠΏΡΠΎΡΠ²ΠΈΠ» ΡΠ΅Π±Ρ ΠΊΠ°ΠΊ ΠΏΠΎΡΠ΅Π½ΡΠΈΠ°Π»ΡΠ½ΠΎΠ΅ ΠΏΡΠΎΡΠΈΠ²ΠΎΠΎΠΏΡΡ
ΠΎΠ»Π΅Π²ΠΎΠ΅ ΡΡΠ΅Π΄ΡΡΠ²ΠΎ
ΠΏΡΠΎΡΠΈΠ² ΠΊΠ»Π΅ΡΠΎΠΊ Π»Π΅ΠΉΠΊΠΎΠ·Π° ΡΠ΅Π»ΠΎΠ²Π΅ΠΊΠ° HL-60 Ρ Π²ΡΡΠ°ΠΆΠ΅Π½Π½ΡΠΌ ΡΠΈΡΠΎΡΡΠ°ΡΠΈΡΠ΅ΡΠΊΠΈΠΌ ΠΈ ΠΏΡΠΎΠ°ΠΏΠΎΠΏΡΠΈΡΠ΅ΡΠΊΠΈΠΌ Π΄Π΅ΠΉΡΡΠ²ΠΈΠ΅ΠΌ
Antiproliferative activity and apoptosis induced by 6-bromo-2-(morpholin-1-yl)-4-anilinoquinazoline on cells of leukemia lines
Quinazolines are known to be multitarget agents with broad spectrum of biological activity. Aim: To investigate anticancer activity of newly prepared 6-bromo-2-(morpholin-1-yl)-4-anilinoquinazoline (BMAQ) towards L1210, HL-60 and U-937 leukemia cells. Materials and Methods: Growth inhibition of BMAQ-treated cells was determined by cell counting using trypan blue staining technique. Apoptosis and cell cycle profile changes were analysed using internucleosomal DNA fragmentation assay, fluorescence microscopy and flow cytometry. Activity of caspase-3 was determined using colorimetric method. Results: Cell proliferation assay showed that BMAQ caused significant decrease of cell number in a dose-dependent manner. BMAQ induced cell death by apoptosis, based on results from DNA fragmentation, fluorescence microscopy and caspase-3 assays. Conclusion: Presented results clearly demonstrate that BMAQ is a promising anticancer agent with significant antiproliferative and apoptotic activities towards leukemia cells in vitro.ΠΠ²ΠΈΠ½Π°Π·ΠΎΠ»ΠΈΠ½Ρ ΠΈΠ·Π²Π΅ΡΡΠ½Ρ ΠΊΠ°ΠΊ Ρ
ΠΈΠΌΠΈΠΎΠΏΡΠ΅ΠΏΠ°ΡΠ°ΡΡ ΡΠΈΡΠΎΠΊΠΎΠ³ΠΎ ΡΠΏΠ΅ΠΊΡΡΠ° Π΄Π΅ΠΉΡΡΠ²ΠΈΡ. Π¦Π΅Π»Ρ: Π½Π° ΠΌΠΎΠ΄Π΅Π»ΡΡ
Π»Π΅ΠΉΠΊΠΎΠ·Π½ΡΡ
ΠΊΠ»Π΅ΡΠΎΠΊ Π»ΠΈΠ½ΠΈΠΉ L1210,
HL-60 ΠΈ U-937 ΠΈΠ·ΡΡΠΈΡΡ ΠΏΡΠΎΡΠΈΠ²ΠΎΠΎΠΏΡΡ
ΠΎΠ»Π΅Π²ΡΡ Π°ΠΊΡΠΈΠ²Π½ΠΎΡΡΡ Π½ΠΎΠ²ΠΎΠ³ΠΎ ΠΏΡΠ΅ΠΏΠ°ΡΠ°ΡΠ° 6-Π±ΡΠΎΠΌΠΎ-2-(ΠΌΠΎΡΡΠΎΠ»ΠΈΠ½-1-ΠΈΠ»)-4-Π°Π½Π°Π»ΠΈΠ½ΠΎΠΈΠ½Π°Π·ΠΎΠ»ΠΈΠ½Π°
(BMAQ). ΠΠ΅ΡΠΎΠ΄Ρ: ΠΈΠ½Π³ΠΈΠ±ΠΈΡΠΎΠ²Π°Π½ΠΈΠ΅ ΡΠΎΡΡΠ° ΠΊΠ»Π΅ΡΠΎΠΊ ΠΏΠΎΠ΄ Π΄Π΅ΠΉΡΡΠ²ΠΈΠ΅ΠΌ BMAQ ΠΈΠ·ΡΡΠ°Π»ΠΈ ΠΏΡΡΠ΅ΠΌ ΠΏΠΎΠ΄ΡΡΠ΅ΡΠ° ΠΊΠΎΠ»ΠΈΡΠ΅ΡΡΠ²Π° ΠΊΠ»Π΅ΡΠΎΠΊ,
ΠΎΠΊΡΠ°ΡΠ΅Π½Π½ΡΡ
ΡΡΠΈΠΏΠ°Π½ΠΎΠ²ΡΠΌ ΡΠΈΠ½ΠΈΠΌ. ΠΠΏΠΎΠΏΡΠΎΠ· ΠΈ ΠΈΠ·ΠΌΠ΅Π½Π΅Π½ΠΈΡ ΠΏΡΠΎΡΠΈΠ»Ρ ΠΊΠ»Π΅ΡΠΎΡΠ½ΠΎΠ³ΠΎ ΡΠΈΠΊΠ»Π° ΠΈΡΡΠ»Π΅Π΄ΠΎΠ²Π°Π»ΠΈ Ρ ΠΏΠΎΠΌΠΎΡΡΡ ΡΠ»ΡΠΎΡΠ΅ΡΡΠ΅Π½ΡΠ½ΠΎΠΉ
ΠΌΠΈΠΊΡΠΎΡΠΊΠΎΠΏΠΈΠΈ, ΡΠ»Π΅ΠΊΡΡΠΎΡΠΎΡΠ΅Π·Π° ΠΠΠ ΠΈ ΠΏΡΠΎΡΠΎΡΠ½ΠΎΠΉ ΡΠΈΡΠΎΠΌΠ΅ΡΡΠΈΠΈ. ΠΠΊΡΠΈΠ²Π½ΠΎΡΡΡ ΠΊΠ°ΡΠΏΠ°Π·Ρ-3 ΠΎΠΏΡΠ΅Π΄Π΅Π»ΡΠ»ΠΈ ΠΊΠΎΠ»ΠΎΡΠΈΠΌΠ΅ΡΡΠΈΡΠ΅ΡΠΊΠΈΠΌ
ΠΌΠ΅ΡΠΎΠ΄ΠΎΠΌ. Π Π΅Π·ΡΠ»ΡΡΠ°ΡΡ: ΠΏΠΎΠΊΠ°Π·Π°Π½ΠΎ, ΡΡΠΎ BMAQ Π²ΡΠ·ΡΠ²Π°Π΅Ρ Π·Π½Π°ΡΠΈΡΠ΅Π»ΡΠ½ΠΎΠ΅ Π΄ΠΎΠ·ΠΎΠ·Π°Π²ΠΈΡΠΈΠΌΠΎΠ΅ ΡΠΌΠ΅Π½ΡΡΠ΅Π½ΠΈΠ΅ ΠΊΠΎΠ»ΠΈΡΠ΅ΡΡΠ²Π° Π»Π΅ΠΉΠΊΠΎΠ·Π½ΡΡ
ΠΊΠ»Π΅ΡΠΎΠΊ. ΠΡΠΈ ΡΡΠΎΠΌ ΠΊΠ»Π΅ΡΠΊΠΈ, ΠΎΠ±ΡΠ°Π±ΠΎΡΠ°Π½Π½ΡΠ΅ BMAQ, ΠΏΠΎΠ³ΠΈΠ±Π°ΡΡ ΠΏΡΡΠ΅ΠΌ Π°ΠΏΠΎΠΏΡΠΎΠ·Π°, ΡΡΠΎ Π΄Π°Π΅ΡΡΡ ΠΎΠ±ΡΠ°Π·ΠΎΠ²Π°Π½ΠΈΠ΅ΠΌ Π°ΠΏΠΎΠΏΡΠΎΡΠΈΡΠ΅ΡΠΊΠΈΡ
ΡΠ΅Π»Π΅Ρ, ΠΌΠ΅ΠΆΠ½ΡΠΊΠ»Π΅ΠΎΡΠΎΠΌΠ½ΠΎΠΉ ΡΡΠ°Π³ΠΌΠ΅Π½ΡΠ°ΡΠΈΠ΅ΠΉ ΠΠΠ ΠΈ Π°ΠΊΡΠΈΠ²Π°ΡΠΈΠ΅ΠΉ ΠΊΠ°ΡΠΏΠ°Π·Ρ-3. ΠΡΠ²ΠΎΠ΄Ρ: ΠΏΡΠ΅Π΄ΡΡΠ°Π²Π»Π΅Π½Π½ΡΠ΅ ΡΠ΅Π·ΡΠ»ΡΡΠ°ΡΡ
ΡΠ²ΠΈΠ΄Π΅ΡΠ΅Π»ΡΡΡΠ²ΡΡΡ ΠΎ ΡΠΎΠΌ, ΡΡΠΎ BMAQ ΠΎΠ±Π»Π°Π΄Π°Π΅Ρ Π°Π½ΡΠΈΠΏΡΠΎΠ»ΠΈΡΠ΅ΡΠ°ΡΠΈΠ²Π½ΠΎΠΉ ΠΈ ΠΏΡΠΎΠ°ΠΏΠΎΠΏΡΠΎΡΠΈΡΠ΅ΡΠΊΠΎΠΉ Π°ΠΊΡΠΈΠ²Π½ΠΎΡΡΡΡ Π² ΠΎΡΠ½ΠΎΡΠ΅Π½ΠΈΠΈ
Π»Π΅ΠΉΠΊΠΎΠ·Π½ΡΡ
ΠΊΠ»Π΅ΡΠΎΠΊ in vitro
Antiproliferative and Proapoptotic Activities of Methanolic Extracts from Ligustrum vulgare L. as an Individual Treatment and in Combination with Palladium Complex
The aim of this study is to examine the growth inhibitory effects of methanolic leaf and fruit extracts of L. vulgare on HCT-116 cells over different time periods and their synergistic effect with a Pd(apox) complex. The antiproliferative activity of plant extracts alone or in combination with the Pd(apox) complex was determined using MTT cell viability assay, where the IC50 value was used as a parameter of cytotoxicity. Results show that antiproliferative effects of L. vulgare extracts increase with extension of exposure time, with decreasing IC50 values, except for 72 h where the IC50 values for methanolic leaf extract were lower than for the fruit extract. The Pd(apox) complex alone had a weak antiproliferative effect, but combination with L. vulgare extracts caused stronger effects with lower IC50 values than with L. vulgare extracts alone. The type of cell death was explored by fluorescence microscopy using the acridin orange/ethidium bromide method. Treatments with plant extracts caused typical apoptotic morphological changes in HCT-116 cells and co-treatments with Pd(apox) complex caused higher levels of apoptotic cells than treatment with plant extracts alone. The results indicate that L. vulgare is a considerable source of natural bioactive substances with antiproliferative activity on HCT-116 cells and which have a substantial synergistic effect with the Pd(apox) complex
Novel insight on the impact of enzymatic addition on organic loading rate in anaerobic digestion
Addition of enzymes to anaerobic digesters (ADs) has been reported as beneficial to the hydrolytic step of the process. Additional benefits have been described for bioadded reactors such as improved dewatering and lower energy requirements. This work aimed to assess the long-term and unaccounted effects of enzymatic addition on sludge digestion. Enzymesβ impacts were tested using different addition modes (bulk or gradual addition) and during operational changes on reactors operated for 295 days. Enzyme added in bulk, generated a 14% increase in biogas production (144 ml/gVSadded) compared to control (126 ml/gVSadded), whereas the same amount of enzyme added gradually produced a 10% increase (139 ml/gVSadded). These values however, where higher when the OLR was increase from 3 to 5.5 kg VS/(m3 day): 257, 212 and 149 ml/gVSadded for the enzyme added in bulk, the enzyme added gradually and the control respectively. Specific biogas yields (SBY), higher in bioadded reactors, were significantly different between control reactors and those reactors dosed in bulk. Furthermore, following OLR increase, the mode of enzyme addition resulted in different increases in gas production rate (GPR) when the enzyme was added in one dose compared to control and to a gradual addition, 121%, 32% and 93% respectively. These results offer a new hypothesis on the impact of bioadditions to AD during changing operational conditions, suggesting a potential stabilising effect of the enzymes in continuous systems
Synthesis and Anticancer Activity of Novel 9-<i>O</i>-Substituted Berberine Derivatives
Berberine is a bioactive isoquinoline alkaloid derived from many plants. Although berberine has been shown to inhibit growth and induce apoptosis of several tumor cell lines, its poor absorption and moderate activity hamper its full therapeutic potential. Here, we describe the synthesis of a series of 9-O-substituted berberine derivatives with improved antiproliferative and apoptosis-inducing activities. An analysis of novel berberine derivatives by EPR spectroscopy confirmed their similar photosensitivity and analogous behavior upon UVA irradiation as berberine, supporting their potential to generate ROS. Improved antitumor activity of novel berberine derivatives was revealed by MTT assay, by flow cytometry and by detection of apoptotic DNA fragmentation and caspase-3 activation, respectively. We showed that novel berberine derivatives are potent inhibitors of growth of HeLa and HL-60 tumor cell lines with IC50 values ranging from 0.7 to 16.7 µM for HL-60 cells and 36 to >200 µM for HeLa cells after 48 h treatment. Further cell cycle analysis showed that the observed inhibition of growth of HL-60 cells treated with berberine derivatives was due to arresting these cells in the G2/M and S phases. Most strikingly, we found that berberine derivative 3 (9-(3-bromopropoxy)-10-methoxy-5,6-dihydro-[1,3]dioxolo[4,5-g]isoquino[3,2-a] isoquinolin-7-ylium bromide) possesses 30-fold superior antiproliferative activity with an IC50 value of 0.7 µM and 6-fold higher apoptosis-inducing activity in HL-60 leukemia cells compared to berberine. Therefore, further studies are merited of the antitumor activity in leukemia cells of this berberine derivative
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