scholarly journals Viability Selection of Bovine Oocyte Using Annexin-V Assay

2013 ◽  
Vol 5 (6) ◽  
Author(s):  
Khairul Osman ◽  
Nurhaslina Hassan ◽  
Siti Fatimah Ibrahim ◽  
Chew Fang Nang ◽  
Zawawi Ismail
Keyword(s):  
Annexin V ◽  
Bovine Oocyte ◽  
Viability Selection ◽  
Annexin V Assay ◽  
Selection Of

scholarly journals The inhibition of ABCB1/MDR1 or ABCG2/BCRP enables doxorubicin to eliminate liver cancer stem cells

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Wang Yin ◽  
Dongxi Xiang ◽  
Tao Wang ◽  
Yumei Zhang ◽  
Cuong V. Pham ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cancer Stem Cells ◽  
Liver Cancer ◽  
High Performance ◽  
Annexin V ◽  
Intracellular Concentration ◽  
Liver Cancer Stem Cells ◽  
Chemotherapy Agent ◽  
Future Strategy ◽  
Annexin V Assay

AbstractTwo ATP-binding cassette transporters, ABCB1/MDR1 and ABCG2/BCRP, are considered the most critical determinants for chemoresistance in hepatocellular carcinoma. However, their roles in the chemoresistance in liver cancer stem cells remain elusive. Here we explored the role of inhibition of MDR1 or ABCG2 in sensitizing liver cancer stem cells to doxorubicin, the most frequently used chemotherapeutic agent in treating liver cancer. We show that the inhibition of MDR1 or ABCG2 in Huh7 and PLC/PRF/5 cells using either pharmacological inhibitors or RNAi resulted in the elevated level of intracellular concentration of doxorubicin and the accompanied increased apoptosis as determined by confocal microscopy, high-performance liquid chromatography, flow cytometry, and annexin V assay. Notably, the inhibition of MDR1 or ABCG2 led to the reversal of the chemoresistance, as evident from the enhanced death of the chemoresistant liver cancer stem cells in tumorsphere-forming assays. Thus, the elevation of effective intracellular concentration of doxorubicin via the inhibition of MDR1 or ABCG2 represents a promising future strategy that transforms doxorubicin from a traditional chemotherapy agent into a robust killer of liver cancer stem cells for patients undergoing transarterial chemoembolization.

scholarly journals A real-time, bioluminescent annexin V assay for the assessment of apoptosis

APOPTOSIS ◽  
2018 ◽  
Vol 24 (1-2) ◽  
pp. 184-197 ◽  
Author(s):  
Kevin Kupcho ◽  
John Shultz ◽  
Robin Hurst ◽  
Jim Hartnett ◽  
Wenhui Zhou ◽  
...  
Keyword(s):  
Real Time ◽  
Annexin V ◽  
Annexin V Assay

scholarly journals The maternal cytoplasmic environment may be involved in the viability selection of gametes and zygotes

Heredity ◽  
2012 ◽  
Vol 110 (4) ◽  
pp. 331-337 ◽  
Author(s):  
Z X Tang ◽  
X F Wang ◽  
M Z Zhang ◽  
Y H Zhang ◽  
D X Deng ◽  
...  
Keyword(s):  
Viability Selection ◽  
Selection Of

A novel approach for the selection of human sperm using annexin V-binding and flow cytometry

2009 ◽  
Vol 91 (4) ◽  
pp. 1285-1292 ◽  
Author(s):  
Christiaan F. Hoogendijk ◽  
Theunis F. Kruger ◽  
Patric J.D. Bouic ◽  
Ralf R. Henkel
Keyword(s):  
Flow Cytometry ◽  
Annexin V ◽  
Human Sperm ◽  
Novel Approach ◽  
Selection Of

TEGDMA Causes Apoptosis in Primary Human Gingival Fibroblasts

2003 ◽  
Vol 82 (10) ◽  
pp. 814-818 ◽  
Author(s):  
V. Janke ◽  
N. von Neuhoff ◽  
B. Schlegelberger ◽  
G. Leyhausen ◽  
W. Geurtsen
Keyword(s):  
Cell Death ◽  
Light Microscopy ◽  
Annexin V ◽  
In Vivo Studies ◽  
Dependent Manner ◽  
Persistent Inflammation ◽  
Proliferation And Apoptosis ◽  
Annexin V Assay ◽  
Triethyleneglycol Dimethacrylate

Previous in vivo studies have revealed that resins may generate a persistent inflammation of oral tissues and cell death as well. Apoptosis is an important regulated process that results in rapid cell death. This study tested the hypothesis that the comonomer triethyleneglycol-dimethacrylate (TEGDMA) causes apoptosis. The effects of TEGDMA on proliferation and apoptosis in primary oral fibroblasts were analyzed by light microscopy and flow cytometry (FACS; Annexin V-assay). TEGDMA at 5 and 7.5 mM inhibited proliferation after 24 hrs. No increased frequency of apoptosis or necrosis was observed with 1 mM or 2.5 mM TEGDMA after 24 hrs. Apoptosis and Annexin V-positive cells were observed with 5 mM and 7.5 mM TEGDMA by light microscopy after 24 hrs. A dramatic increase in apoptotic cells was detected by FACS after 24 hrs with 7.5 mM TEGDMA. Thus, TEGDMA was cytotoxic and “apoptotic” in a dose- and time-dependent manner.

scholarly journals Biological Evaluation of Arylsemicarbazone Derivatives as Potential Anticancer Agents

2019 ◽  
Vol 12 (4) ◽  
pp. 169
Author(s):  
Anne Cecília Nascimento da Cruz ◽  
Dalci José Brondani ◽  
Temístocles I´talo de Santana ◽  
Lucas Oliveira da Silva ◽  
Elizabeth Fernanda da Oliveira Borba ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Cell Lines ◽  
Anticancer Agents ◽  
Cell Cycle Progression ◽  
Annexin V ◽  
Biological Evaluation ◽  
G1 Phase ◽  
Cellular Functions ◽  
Ic50 Values ◽  
Annexin V Assay

Fourteen arylsemicarbazone derivatives were synthesized and evaluated in order to find agents with potential anticancer activity. Cytotoxic screening was performed against K562, HL-60, MOLT-4, HEp-2, NCI-H292, HT-29 and MCF-7 tumor cell lines. Compounds 3c and 4a were active against the tested cancer cell lines, being more cytotoxic for the HL-60 cell line with IC50 values of 13.08 μM and 11.38 μM, respectively. Regarding the protein kinase inhibition assay, 3c inhibited seven different kinases and 4a strongly inhibited the CK1δ/ε kinase. The studied kinases are involved in several cellular functions such as proliferation, migration, cell death and cell cycle progression. Additional analysis by flow cytometry revealed that 3c and 4a caused depolarization of the mitochondrial membrane, suggesting apoptosis mediated by the intrinsic pathway. Compound 3c induced arrest in G1 phase of the cell cycle on HL-60 cells, and in the annexin V assay approximately 50% of cells were in apoptosis at the highest concentration tested (26 μM). Compound 4a inhibited cell cycle by accumulation of abnormal postmitotic cells at G1 phase and induced DNA fragmentation at the highest concentration (22 μM).

Carvacrol Induced Program Cell Death and Cell Cycle Arrest in Androgen-Independent Human Prostate Cancer Cells via Inhibition of Notch Signaling

2019 ◽  
Vol 19 (13) ◽  
pp. 1588-1608 ◽  
Author(s):  
Fahad Khan ◽  
Vipendra K. Singh ◽  
Mohd Saeed ◽  
Mohd A. Kausar ◽  
Irfan A. Ansari
Keyword(s):  
Prostate Cancer ◽  
Cell Cycle ◽  
Notch Signaling ◽  
Cancer Cells ◽  
Prostate Cancer Cell ◽  
Cancer Cell Line ◽  
Annexin V ◽  
Prostate Cancer Cell Line ◽  
Prostate Cancer Cells ◽  
Annexin V Assay

Background: Several studies have revealed that abnormal activation of Notch signaling is closely related with the development and progression of prostate cancer. Although there are numerous therapeutic strategies, a more effective modality with least side effects is urgently required for the treatment of prostate cancer. Carvacrol is a monoterpenoid phenol and majorly present in the essential oils of Lamiaceae family plants. Many previous reports have shown various biological activities of carvacrol like antioxidant, antiinflammatory and anticancer properties. Recently, we have shown potent anticancer property of carvacrol against prostate cancer cell line DU145. In the current study, we report the chemopreventive and therapeutic potential of carvacrol against another prostate cancer cell line PC-3 with its detailed mechanism of action. Methods: To determine the effect of the carvacrol on prostate cancer cells, the cell viability was estimated by MTT assay and cell death was estimated by LDH release assay. The apoptotic assay was performed by DAPI staining and FITC-Annexin V assay. Reactive Oxygen Species (ROS) was estimated by DCFDA method. Cell cycle analysis was performed by flow cytometry. Gene expression analysis was performed by quantitative real time PCR. Results: Our results suggested that the carvacrol treatment significantly reduced the cell viability of PC-3 cells in a dose- and time-dependent manner. The antiproliferative action of carvacrol was correlated with apoptosis which was confirmed by nuclear condensation, FITC-Annexin V assay, modulation in expression of Bax, Bcl-2 and caspase activation. The mechanistic insight into carvacrol-induced apoptosis leads to finding of elevated level of Reactive Oxygen Species (ROS) and mitochondrial membrane potential disruption. Cell cycle analysis revealed that carvacrol prevented cell cycle in G0/G1 that was associated with decline in expression of cyclin D1 and Cyclin-Dependent Kinase 4 (CDK4) and augmented expression of CDK inhibitor p21. Having been said the role of hyperactivation of Notch signaling in prostate cancer, we also deciphered that carvacrol could inhibit Notch signaling in PC-3 cells via downregulation of Notch-1, and Jagged-1. Conclusion: Thus, our previous and current findings have established the strong potential of carvacrol as a chemopreventive agent against androgen-independent human prostate cancer cells.

Monocytic Differentiation of Myeloid Leukemia Cell Lines Induced by ATRA and 5-Aza-2'-Deoxycitidine.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 3111-3111
Author(s):  
Atsushi Fujiki ◽  
Toshihiko Imamura ◽  
Yoshifumi Hirashima ◽  
Mitsuru Miyachi ◽  
Shigeki Yagyu ◽  
...  
Keyword(s):  
Growth Inhibition ◽  
Cell Lines ◽  
Myeloid Leukemia ◽  
Annexin V ◽  
G1 Arrest ◽  
Inhibition Rate ◽  
Monocytic Differentiation ◽  
Differentiation Therapy ◽  
Specific Pcr ◽  
Annexin V Assay

Abstract Abstract 3111 Poster Board III-48 Backgrounds and Introduction The CCAAT/enhancer binding protein (C/EBP)αa is a transcriptional factor of hematopoietic system and plays a key role in monocytic differentiation. Recently, several studies have reported that C/EBPαa expression is down-regulated in acute myeloid leukemia (AML), leading to block of granulocytic and monocytic differentiation. Differentiation therapy of ATRA is highly effective for acute promyelocytic leukemia (APL). The mechanism of induction of differentiation in the treatment of APL is induction of a set of transcriptional factors which are responsible for myeloid differentiation. However, ATRA alone is not sufficient to treat another type of AML. Thus, it is worth to explore the agents which intensify the efficacy of ATRA. To assess the possibility of differentiation therapy in AML, except for APL, we evaluated the efficacy of demethylation agent combined with ATRA for various AML cell lines. Materials and Methods The five AML cell lines (K562, U-937, HL-60, THP-1, and KOCL48 expressing MLL-AF4) were treated with 50nM 5-Aza-2'-deoxycytidine (5-Aza) for two days and 1 mM ATRA for additional five days. Then, we analyzed cell growth with counting nuclei using Coulter counter. The cell cycle analysis was also performed by flow cytometry (FCM). In addition, Annexin V assay was performed to determine whether apoptosis occurred or not. To assess whether monocytic differentiation was induced or not, the expression of CD11b was evaluated by FCM. In addition, the expression of transcriptional factors, such as C/EBPαa, PU.1 and c-myc ) were analyzed by real time PCR analysis. Methylation specific PCR was also performed to evaluate the methylation status of promoter region of C/EBPαa. Results HL-60 was highly sensitive to ATRA (growth inhibition rate: 80%). THP-1 and KOCL-48 were moderately sensitive to ATRA (growth inhibition rate: 50-60%). Addition of 5-Aza induced suppression of the growth in these two cell lines efficiently (growth inhibition rate: 80%). K562 and U937 were resistant to ATRA (growth inhibition rate: 10-20%). Addition of 5-Aza induced suppression of cell-growth in U937 (growth inhibition rate: 70%). However, 5-Aza did not show the effect in K562. Morphological studies revealed the characteristic features, such as extended cytoplasm with vacuoles, fine granules and irregular shaped nucleus, were evident in four cell lines which was sensitive to 5-Aza. FCM analysis revealed intensification of CD11b expression. In addition, real time PCR determined the increased expression of C/EBPαa and PU.1 (Fold change: 2-3.0) in the four cell lines except for K562. On the other hand, the expression level of c-myc was decreased under treatment with ATRA and 5-Aza (Fold change: 0.2). Cell cycle analysis revealed G1 arrest was occurred. Annexin V assay also revealed that combination therapy of ATRA and 5-Aza induced apoptosis in theses cell lines except for K562. Methylation specific PCR did not identified hypermetylation of pormorter region of C/EBPαa in these cell lines except for K562. Conclusion Addition of 5-Aza to ATRA induced further expression of C/EBPαa and PU.1 efficiently, leading to monocytic differentiation in AML cell lines. Monocytic differentiation was accompanied with G1 arrest through down-regulation of c-myc, and apoptosis was induced finally. Combination of ATRA and 5-Aza might be effective therapeutic option even for AML which is resistant to differentiation therapy with ATRA. Disclosures No relevant conflicts of interest to declare.

scholarly journals The Effect of Lycopene Preexposure on UV-B-Irradiated Human Keratinocytes

2016 ◽  
Vol 2016 ◽  
pp. 1-15 ◽  
Author(s):  
Andreia Ascenso ◽  
Tiago Pedrosa ◽  
Sónia Pinho ◽  
Francisco Pinho ◽  
José Miguel P. Ferreira de Oliveira ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Annexin V ◽  
Human Keratinocytes ◽  
Protective Role ◽  
Cell Cycle Distribution ◽  
Skin Cells ◽  
A Cell ◽  
Previously Treated ◽  
Annexin V Assay ◽  
And Oxidative Stress

Lycopene has been reported as the antioxidant most quickly depleted in skin upon UV irradiation, and thus it might play a protective role. Our goal was to investigate the effects of preexposure to lycopene on UV-B-irradiated skin cells. Cells were exposed for 24 h to 10 M lycopene, and subsequently irradiated and left to recover for another 24 h period. Thereafter, several parameters were analyzed by FCM and RT-PCR: genotoxicity/clastogenicity by assessing the cell cycle distribution; apoptosis by performing the Annexin-V assay and analyzing gene expression of apoptosis biomarkers; and oxidative stress by ROS quantification. Lycopene did not significantly affect the profile of apoptotic, necrotic and viable cells in nonirradiated cells neither showed cytostatic effects. However, irradiated cells previously treated with lycopene showed an increase in both dead and viable subpopulations compared to nonexposed irradiated cells. In irradiated cells, lycopene preexposure resulted in overexpression ofBAXgene compared to nonexposed irradiated cells. This was accompanied by a cell cycle delay at S-phase transition and consequent decrease of cells in G0/G1 phase. Thus, lycopene seems to play a corrective role in irradiated cells depending on the level of photodamage. Thus, our findings may have implications for the management of skin cancer.

Anti-Tumour Effect of Zoledronate in Cells from B Chronic Lymphoytic Leukemia and Low-Grade Lymphoma Patients.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 4830-4830
Author(s):  
Deborah Vidal Vasconcellos ◽  
Karina Lani Silva ◽  
Eric Delfraro de Paula Castro ◽  
Arthur Coelho Moellman ◽  
Maria do Socorro Pombo-de-Oliveira ◽  
...  
Keyword(s):  
Annexin V ◽  
Previous Treatment ◽  
Lymphocytic Leukemia ◽  
Low Grade ◽  
Complementary Role ◽  
Apoptotic Effect ◽  
Previously Treated ◽  
Annexin V Assay ◽  
Anti Tumour Effect ◽  
In Cells

Abstract Recent studies demonstrate that bisphosphonates - anti-resorptive drugs - have direct anti-tumour effect in many tumour cell lines, including hematopoietic ones. The aim of this study was to evaluate the apoptotic effect of Zoledronate in cells from B chronic lymphocytic leukemia (B-CLL) and low-grade lymphoma (LGL) patients. Samples of 19 B-CLL or LGL patients were incubated with Zoledronate 100 μM in RPMI medium supplemented with fetal bovine serum 10% at 370C for 12h. Apoptosis using Annexin V assay, by flow cytometry (FC), was observed in 8 out of 19 (42,1%) patients despite previous treatment. Multidrug resistance (MDR) phenotype was performed using rhodamine-123 efflux assay by FC. Our results demonstrate that Zoledronate 100 m M can induce apoptosis in B malignant lymphocytes despite previous treatment and MDR phenotype. So, patients previously treated with distinct therapeutic agents or not can potentially benefit from the anti-tumour effect of Zoledronate. This is the first work demonstrating the anti-tumour effect of Zoledronate in newly obtained cells from patients with B-CLL and LGL. These results in association with evidences from recent studies suggest that Zoledronate may have an important and complementary role in hematological malignant diseases.

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