IN VITRO DETERMINATION OF THE APOPTOTIC EFFECT OF HEPARIN ON LYMPHOBLASTS USING DNA ANALYSIS AND MEASUREMENTS OF Fas AND Bcl-2 PROTEINS BY FLOW CYTOMETRY

2004 ◽  
Vol 21 (5) ◽  
pp. 383-391 ◽  
Author(s):  
Erol Erduran ◽  
Yavuz Tekelioglu ◽  
Yusuf Gedik ◽  
İsmail Bektaş ◽  
Sadan Hacisalihoglu
Keyword(s):  
Flow Cytometry ◽  
Dna Analysis ◽  
Apoptotic Effect

Effect of reticulocytosis on lactate dehydrogenase isoenzyme distribution in serum: in vivo and in vitro studies

1990 ◽  
Vol 36 (9) ◽  
pp. 1638-1641 ◽  
Author(s):  
S C Kazmierczak ◽  
W J Castellani ◽  
F Van Lente ◽  
E D Hodges ◽  
B Udis
Keyword(s):  
Flow Cytometry ◽  
Lactate Dehydrogenase ◽  
Cell Types ◽  
In Vitro Studies ◽  
Hemolytic Disease ◽  
Dehydrogenase Isoenzyme ◽  
Disease Analysis

Abstract We investigated the effect of reticulocytosis on the lactate dehydrogenase (LD; EC 1.1.1.27) isoenzyme LD1/LD2 ratio in patients with and without evidence of hemolytic disease. Analysis of sera from patients with reticulocytosis and in vivo hemolysis showed a mean LD1/LD2 ratio of 0.92 compared with a ratio of 0.69 in patients with in vivo hemolysis and normal reticulocyte counts. Determination of LD isoenzymes in erythrocyte lysate revealed significantly increased LD1/LD2 ratios for patients with marked reticulocytosis compared with those for patients with normal-to-minimal increases in reticulocytes. Finally, separation of mature erythrocytes and reticulocytes by flow cytometry revealed marked differences in the LD1/LD2 isoenzyme distribution between these two cell types. The ability of hemolysis to cause a "flipped" LD1/LD2 ratio is dependent on the proportion of the hemolyzed cells that are reticulocytes.

scholarly journals Upregulation of pERK and c-JUN by γ-tocotrienol and not α-tocopherol are essential to the differential effect on apoptosis in prostate cancer cells.

2020 ◽  
Author(s):  
Christine Moore ◽  
Victoria Palau ◽  
Rashid Mahboob ◽  
Janet Lightner ◽  
William Stone ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Cancer Cells ◽  
Prostate Cancer Cells ◽  
Cytotoxic Effects ◽  
Jnk Pathway ◽  
Androgen Sensitivity ◽  
Apoptotic Effect ◽  
Select Trial

Abstract Background: α-tocopherol (AT) and γ-tocotrienol (GT3) are vitamin E isoforms considered to have potential chemopreventive properties. AT has been widely studied in vitro and in clinical trials with mixed results. The latest clinical study (SELECT trial) tested AT in prostate cancer patients, determined that AT provided no benefit, and could promote cancer. Conversely, GT3 has shown antineoplastic properties in several in vitro studies, with no clinical studies published to date. GT3 causes apoptosis via upregulation of the JNK pathway; however, inhibition results in a partial block of cell death. We compared side by side the mechanistic differences in these cells in response to AT and GT3.Methods: The effects of GT3 and AT were studied on androgen sensitive LNCaP and androgen independent PC-3 prostate cancer cells. Their cytotoxic effects were analyzed via MTT and confirmed by metabolic assays measuring ATP. Cellular pathways were studied by immunoblot. Quantitative analysis and the determination of relationships between cell signaling events were analyzed for both agents tested. Non-cancerous prostate RWPE-1 cells were also included as a control. Results: The RAF/RAS/ERK pathway was significantly activated by GT3 in LNCaP and PC-3 cells but not by AT. This activation is essential for the apoptotic affect by GT3 as demonstrated the complete inhibition of apoptosis by MEK1 inhibitor U0126. Phospho-c-JUN was upregulated by GT3 but not AT. No changes were observed on AKT for either agent, and no release of cytochrome c into the cytoplasm was detected. Caspases 9 and 3 were efficiently activated by GT3 on both cell lines irrespective of androgen sensitivity, but not in cells dosed with AT. Cell viability of non-cancerous RWPE-1 cells was affected neither by GT3 nor AT. Conclusions: c-JUN is a recognized master regulator of apoptosis as shown previously in prostate cancer. However, the mechanism of action of GT3 in these cells also include a significant activation of ERK which is essential for the apoptotic effect of GT3. The activation of both, ERK and c-JUN, is required for apoptosis and may suggest a relevant step in ensuring circumvention of mechanisms of resistance related to the constitutive activation of MEK1.

Comparison the effects of Sambucus ebulus leaf and fruit extracts on Leishmania major in-vitro

2019 ◽  
Vol 20 ◽  
Author(s):  
Samira Kadkhodamasoum ◽  
Farahnaz Bineshian ◽  
Amir KarimiPour ◽  
Pooya Tavakoli ◽  
Masoud Foroutan ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Leaf Extract ◽  
Leishmania Major ◽  
Drug Candidate ◽  
Fruit Extract ◽  
Fruit Extracts ◽  
Sambucus Ebulus ◽  
Apoptotic Effect ◽  
Ic50 Concentration

Background;: Leishmaniasis is one of the major diseases caused by the intracellular parasite of Leishmania. It has become one of the most dangerous health problems today. Our aim of the present study is to compare the effects of Sambucus ebulus leaf and fruit extracts on Leishmania major in vitro. Methods: In this study, we used MTT, promastigote and amastigote assay to evaluate the effect of different concentrations of the extract on parasite and we compared their effects. The flow cytometry technique was also used to detect the apoptotic effect of the extracts on promastigotes. Results: According to MTT experiment IC50 concentration of leaf and fruit extracts on parasite was 157 μg/ml and 265 μg/ml, respectively. After analysis by flow cytometry, leaf and fruit extracts also showed apoptosis effect. Leaf and fruit extract caused 40.2 and 2.67 percent apoptosis. Conclusion: Based on the above assessment, we determined that the S. ebulus leaf extract has a more toxic effect on promastigotes and amstigotes than its fruit extract and maybe in the future that be used as a drug candidate.

scholarly journals Upregulation of pERK and c-JUN by γ-tocotrienol and not α-tocopherol are essential to the differential effect on apoptosis in prostate cancer cells.

2020 ◽  
Author(s):  
Christine Moore ◽  
Victoria Palau ◽  
Rashid Mahboob ◽  
Janet Lightner ◽  
William Stone ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Cancer Cells ◽  
Prostate Cancer Cells ◽  
Cytotoxic Effects ◽  
Jnk Pathway ◽  
Androgen Sensitivity ◽  
Apoptotic Effect ◽  
Select Trial

Abstract Background: α-tocopherol (AT) and γ-tocotrienol (GT3) are vitamin E isoforms considered to have potential chemopreventive properties. AT has been widely studied in vitro and in clinical trials with mixed results. The latest clinical study (SELECT trial) tested AT in prostate cancer patients, determined that AT provided no benefit, and could promote cancer. Conversely, GT3 has shown antineoplastic properties in several in vitro studies, with no clinical studies published to date. GT3 causes apoptosis via upregulation of the JNK pathway; however, inhibition results in a partial block of cell death. We compared side by side the mechanistic differences in these cells in response to AT and GT3. Methods: The effects of GT3 and AT were studied on androgen sensitive LNCaP and androgen independent PC-3 prostate cancer cells. Their cytotoxic effects were analyzed via MTT and confirmed by metabolic assays. Cellular pathways were studied by immunoblot. Quantitative analysis and the determination of relationships between cell signaling events were analyzed for both agents tested. Non-cancerous prostate RWPE-1 cells were also included as a control. Results: The RAF/RAS/ERK pathway was significantly activated by GT3 in LNCaP and PC-3 cells but not by AT. This activation is essential for the apoptotic affect by GT3 as demonstrated the complete inhibition of apoptosis by MEK1 inhibitor U0126. Phospho-c-Jun was upregulated by GT3 but not AT. No changes were observed on AKT for either agent, and no release of cytochrome c into the cytoplasm was detected. Caspases 9 and 3 were efficiently activated by GT3 on both cell lines irrespective of androgen sensitivity, but not in cells dosed with AT. Cell viability of non-cancerous RWPE-1 cells was affected neither by GT3 nor AT. Conclusions: c-JUN is a recognized master regulator of apoptosis as shown previously in prostate cancer. However, the mechanism of action of GT3 in these cells also include a significant activation of ERK which is essential for the apoptotic effect of GT3. The activation of both, ERK and c-JUN, is required for apoptosis and may suggest a relevant step in ensuring circumvention of mechanisms of resistance related to the constitutive activation of MEK1.

scholarly journals Cytotoxicity, Antiproliferative and Apoptotic Effect of n-Hexane Fraction of Lime Parasite (Dendrophtoe pentandra)

Molekul ◽  
2019 ◽  
Vol 14 (1) ◽  
pp. 1
Author(s):  
Subandrate Subandrate ◽  
Masayu Farah Diba ◽  
Salni Salni ◽  
Triwani Triwani ◽  
Sri Nita
Keyword(s):  
Flow Cytometry ◽  
Cytotoxic Activity ◽  
Doubling Time ◽  
T47d Cell ◽  
Mtt Method ◽  
Folk Remedy ◽  
Proliferation And Apoptosis ◽  
Cell Doubling Time ◽  
Apoptotic Effect

Breast cancer is one of the biggest causes of death in women in the world. Lime parasite (Dendrophtoe pentandra (L.) Miq.), a folk remedy used by Indonesian people, is believed to be efficacious as anticancer drug. This research aims to know the activity of n-hexane fractions of lime parasite in inhibiting the proliferation and apoptosis of T47D cells in vitro. Cytotoxic test with MTT method assay from n-hexane fractions used a multilevel concentration. Antiproliferative test was carried out by the method of MTT assay and cell doubling time was calculated at the time of duplication. Apoptotic test was done with concentration of 1 IC50and ½ IC50which was analyzed by flow cytometry. The results reveals that fractions of lime parasite have cytotoxic activity with concentration of IC50is included in moderatecytotoxic level. The result of the doubling time of the optimum fraction of n-hexane is in 31 hours with the concentration of ¼IC50. Results for the flow cytometry shows the fraction of n-hexane does not induce apoptosis in cells of T47D.  Those results show that the active fraction of lime parasite has cytotoxic activity which is able to inhibit proliferation, but does not induce apoptosis of T47D cell.

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