Thymoquinone Effects on Cell Viability, Apoptosis and VEGF-A Gene Expression Level in AGS(CRL-1739) Cell Line

2019 ◽  
Vol 19 (6) ◽  
pp. 820-826 ◽  
Author(s):  
Mohsen Rashid ◽  
Forough Sanjarin ◽  
Farzaneh Sabouni
Keyword(s):  
Gene Expression ◽  
Cell Proliferation ◽  
Cell Line ◽  
Real Time ◽  
Mtt Assay ◽  
Nigella Sativa ◽  
Angiogenic Factor ◽  
Cancer Drug ◽  
Anti Cancer ◽  
Ags Cell Line

Background: Cancer is one of the most fatal diseases across the world and it was reported that 90% of cancer fatality depends on its angiogenesis potential. Black seed or Nigella sativa L. is a medicinal plant native to southwest Asia. N. sativa has been used for medicinal purposes for centuries and predominantly has bioactive components like Thymoquinone, which is used as a candidate for anti-cancer and anti-angiogenesis drugs. Methods: Callus was induced from leaf tissue, after that alcoholic extracts were prepared from three-month-old calluses. Thymoquinone content was measured by HPLC methods. AGS cell line was cultured and treated with standard Thymoquinone and extracts from callus. Then, cell proliferation, expression of angiogenic factor (VEGF-A gene), and apoptosis test were done by MTT assay, real-time PCR and Annexin-v kit, respectively. Results: HPLC found the maximum amount of Thymoquinone in the extract of leaf calluses, which were grown in the dark. MTT assay revealed that particular doses of extracts reduced cell proliferation. Real-time and Fluorescence- Activated Cell Sorting (FACS) results demonstrated that standard Thymoquinone and callus extracts down-regulated the VEGF-A gene expression, and all three induced apoptosis in the AGS cell line. Conclusion: It has been shown that TQ has pro-apoptotic and anti-metastatic effects on stomach cancer cell line, and these properties can introduce it as an anti-cancer drug in the near future.

scholarly journals Jianpi Yangwei decoction promotes apoptosis and suppresses proliferation of 5-fluorouracil resistant gastric cancer cells in vitro and in vivo

2020 ◽  
Vol 20 (1) ◽  
Author(s):  
Huijuan Tang ◽  
Wenjie Huang ◽  
Qiang Yang ◽  
Ying Lin ◽  
Yihui Chen ◽  
...  
Keyword(s):  
Gastric Cancer ◽  
Drug Resistance ◽  
Cell Line ◽  
Real Time ◽  
Mtt Assay ◽  
Xenograft Tumor ◽  
Rt Pcr ◽  
Induced Apoptosis

Abstract Background The exploration of new therapeutic agents targeting 5-Fu resistance may open a new opportunity to gastric cancer treatment. The objective is to establish a 5-Fu resistant gastric cancer cell line and observe the effect of Jianpi Yangwei decoction (JPYW) on its apoptosis and drug-resistance related proteins. Methods MTT assay was used to measure the effect of JPYW on the BGC823 cells proliferation, and the apoptosis was observed by flow cytometry and Hoechst fluorescence staining. The BGC823 xenograft tumor nude mice models were established, the apoptosis was detected by Tunel method. BGC-823/5-Fu was established by repeated low-dose 5-Fu shocks, the drug resistance index and proliferation were detected by the MTT assay; MDR1 mRNA was detected by real-time RT-PCR; Western blot was used to detect the ratio of p-AKT to AKT; The BGC823/5-Fu xenograft tumor nude mice models were established and apoptosis was measured. The expressions of MRP1, MDR1, ABCG2, AKT, p-AKT, caspase-3 and bcl-2 were detected by immunohistochemistry and the AKT mRNA expression was detected by real-time RT-PCR. Results JPYW induced apoptosis in BGC823 cells; Drug-resistant cell line BGC-823/5-Fu was sucessfully established; JPYW induced apoptosis of BGC823/5-Fu cells, down-regulated the expression of MRP1, MDR1 and ABCG2 in vitro and in vivo, and further decreased MDR1 expression when combined with pathway inhibitor LY294002 (P < 0.05); JPYW down-regulated the ratio of p-AKT to AKT in vitro in a dose-dependent manner, the same as after the combination with LY294002 (P < 0.05). Conclusion JPYW can induce apoptosis of BGC823 and BGC823/5-Fu cells, and down-regulate the expression of MDR1, MRP1, ABCG2 in vitro and in vivo. Its in vitro effect is related to the PI3K/AKT signaling pathway.

Effects of Phthalates on the Human Corneal Endothelial Cell Line B4G12

2012 ◽  
Vol 31 (4) ◽  
pp. 364-371 ◽  
Author(s):  
Tanja Krüger ◽  
Yi Cao ◽  
Søren K. Kjærgaard ◽  
Lisbeth E. Knudsen ◽  
Eva C. Bonefeld-Jørgensen
Keyword(s):  
Gene Expression ◽  
Cell Proliferation ◽  
Endothelial Cell ◽  
Cell Line ◽  
Corneal Endothelial Cell ◽  
Industrial Chemicals ◽  
Cell Secretion ◽  
Endothelial Cell Line ◽  
Butyl Phthalate

Phthalates are industrial chemicals used in many cosmetics. We evaluated an in vitro model for eye irritancy testing using the human corneal endothelial cell line B4G12. Cell proliferation and toxicity were assessed after exposing to di- n-butyl phthalate (DBP), benzyl butyl phthalate (BBP), di-2-ethylhexyl phthalate (DEHP), diisodecyl phthalate (DIDP), di- n-octyl phthalate (DnOP), and di-isononyl phthalate (DINP). Gene expression and secretion of inflammatory cytokines were evaluated after exposure to DBP. Decreased cell proliferation was observed for the phthalates DBP, BBP, and DEHP, and cell toxicity was observed for DBP and BBP. Upon DBP exposure at nontoxic concentrations, a significant increased gene expression and cytokine cell secretion were observed for interleukin-1β (IL-1β) and IL-8, and also an increased IL-6 secretion was observed. In conclusion, the human corneal endothelial cell line B4G12 may be a potential model for inflammatory eye irritancy testing of phthalates.

scholarly journals Visualizing Gene Expression in Real-Time

2005 ◽  
Vol 13 (3) ◽  
pp. 3-7
Author(s):  
Stephen W. Carmichael
Keyword(s):  
Gene Expression ◽  
Cell Line ◽  
Real Time ◽  
Temporal Resolution ◽  
Protein Sequence ◽  
Living Cells ◽  
Entire Process ◽  
Gene Chips ◽  
A Cell ◽  
Dna Template

Gene expression has been visualized for a few decades, but in static forms such as blots and gene chips. Susan Janicki, Toshiro Tsukamoto, Sim one Salghetti, William Tansey, Ravi Sachidanandam, Kannanganattu Prasanth, Thomas Ried, Yaron Shav-Tal, Edouard Bertrand, Robert Singer, and David Spector have recently designed a cell line in which gene expression can be observed with stunningly accurate spatial and temporal resolution. Gene expression is a cascade of events beginning with transcription of RNA from the DNA template and ending with translation into a protein sequence. Janicki et al. were able to visualize the entire process at the levels of DNA, RNA, and proteins in living cells!

Arginine Vasopressin Inhibition of Cyclin D1 Gene Expression Blocks the Cell Cycle and Cell Proliferation in the Mouse Y1 Adrenocortical Tumor Cell Line†

Biochemistry ◽  
2003 ◽  
Vol 42 (7) ◽  
pp. 2116-2121 ◽  
Author(s):  
Telma T. Schwindt ◽  
Fábio L. Forti ◽  
Maria Ap. Juliano ◽  
Luiz Juliano ◽  
Hugo A. Armelin
Keyword(s):  
Gene Expression ◽  
Cell Cycle ◽  
Cell Proliferation ◽  
Tumor Cell ◽  
Cyclin D1 ◽  
Cell Line ◽  
Arginine Vasopressin ◽  
Tumor Cell Line ◽  
Adrenocortical Tumor

scholarly journals A REVIEW ON DRUG NIMBUKA AS ANTIBACTERIAL, ANTICANCEROUS AND ADJUNCTIVE FOR CHEMOTHERAPY

2020 ◽  
pp. 333-334
Author(s):  
Dr. Naveena K S ◽  
Dr. Shrinath M. Vaidya
Keyword(s):  
Mortality Rate ◽  
Key Words ◽  
Cell Line ◽  
Vital Statistics ◽  
Cancer Drug ◽  
Citrus Limon ◽  
Abnormal Cell ◽  
Anti Cancer ◽  
Citrus Medica

In present scenario Cancer contributes to highest mortality rate. According to the vital statistics of 2016, around 14 lakh deaths have occurred due to Cancer. Cancer is conditions were new growth and division of abnormal cell is going to happen. Nimbuka is the drug explained in Ayurveda by Acharya Bhavaprakasha as Krimigna and Prakruti-sthapaka. Nimbuka is anti-cancerous and adjunctive for chemotherapy and Rasayana (as per the information obtained from cell line studies). Nimbuka belongs to Rutaceae family which itself is proved having the anti-cancer drug property. KEY WORDS: Nimbuka, Anticancerous, Citrus medica, Citrus limon etc

scholarly journals Assessment of the synergic effect of Avastin and VEGF siRNA on inhibition of cell proliferation and angiogenic factor expression of human breast cancer cells

2021 ◽  
Author(s):  
Abdolkhalegh Deezagi ◽  
Bahar Ghorbani
Keyword(s):  
Breast Cancer ◽  
Cell Proliferation ◽  
Cell Death ◽  
Real Time ◽  
Real Time Pcr ◽  
Angiogenic Factor ◽  
Cell Counting ◽  
Synergic Effect ◽  
Vegf Expression ◽  
Cell Growth And Migration

Abstract Angiogenesis is an important process in tumor growth and metastasis and vascular endothelial growth factor (VEGF) plays an important role in this process. Several VEGF inhibitors have been developed as anticancer agents including humanized monoclonal antibodies, and various small molecules. The aim of this work was to investigate the effect of combination of VEGF siRNA and Avastin on breast cancer MCF-7 cell line behavior. For this purpose, the cells were treated with different concentrations of Avastin and/or VEGF siRNA and their combination. The cell survival and cell proliferation were assayed by cell counting, trypan blue and MTT tests. The cell migration was assayed by scratching test. VEGF expression was assayed by RT and real-time PCR and ELISA methods. Results indicated the significant increase in cell death following treatment with Avastin (50% cell death at 100 µg/ml). Cell death with VEGF siRNA transfection was lower than Avastin, however, it was significant. This result in VEGF siRNA + Avastin (100 µg/ml) treatment was greater compared to treatment with each of these compounds alone (47%). Scratching results also showed the synergic effect of VEGF siRNA and Avastin (57% decrease). Real-time PCR results showed that Avastin at concentrations of ≥ 50 µg/ml led to 2.5 to 7.5-fold decrease in VEGF expression levels. Also, treatment with VEGF siRNA led to 15.5-fold decrease in VEGF expression. Finally, VEGF expression following VEGF siRNA + Avastin treatment led to a significant 47.5-fold decrease in VEGF expression. It could be concluded that combination of VEGF siRNA and Avastin have a more significant impact on the inhibition of cell growth and migration and it can probably be used as an effective therapeutic approach.

scholarly journals Dysregulation of ANKRD11 Influenced Hematopoisis By Histone Acetylation-Mediated Gene Expression in Myelodysplastic Syndrome

Blood ◽  
2016 ◽  
Vol 128 (22) ◽  
pp. 4292-4292
Author(s):  
Youshan Zhao ◽  
Feng Xu ◽  
Juan Guo ◽  
Sida Zhao ◽  
Chunkang Chang ◽  
...  
Keyword(s):  
Gene Expression ◽  
Cell Proliferation ◽  
Histone Acetylation ◽  
Mrna Expression ◽  
Cell Line ◽  
Expression Levels ◽  
Target Sequencing ◽  
Cell Proliferation And Differentiation ◽  
Proliferation And Differentiation ◽  
Mrna Expression Levels

Abstract Background and Object In addition to histone deacetylation, the importance of histone over-acetylation induced oncogene transcription in initiation and progression of myelodysplastic syndrome (MDS) has been proposed recently. Our previous whole-exome sequencing identified a new somatic mutation, ANKRD11, an important factor in histone acetylation regulation. Its roles in MDS pathophysiology need to be clarified. Methods The next generation target sequencing (Including ANKRD11) was carried out in 320 patients with MDS using the MiSeq Benchtop Sequencer. ANKRD11 mRNA expression in bone marrow of MDS was measured by real-time PCR. Loss and gain of function assay were carried out in myeloid cell lines K562, MEG-01£¬or SKM-1 to observe the influence on cell proliferation and differentiation . The levels of histone acetylation at H3 and H4 were detected by Western blot. Results Target sequencing in a cohort of 320 MDS patients identified 14 of ANKRD11 mutations (4.38%, Fig.1), which were confirmed by Sanger sequencing. Meanwhile, no ANKRD11 mutations in 100 normal controls were defined. ANKRD11 mutations occurred frequently in exons 10 and 9. The mRNA expression levels of ANKRD11 were significantly decreased in MDS patients, especially in ANKRD11mutant patients (Fig.2). ANKRD11 knockdown in K562 and MEG-1 resulted in growth inhibition, cell cycle arrest and erythroid/megakaryocytic differentiation retardant. In MDS cell line SKM-1, the arrested differentiation was rescued by over-expression of ANKRD11. Consistent with a role for ANKRD11 in histone acetylation, ANKRD11 KD increased acetylation of histones H3 and H4 at H3K14 and H4K5 and resulted in the upregulation of genes involved in differentiation inhibilation (SOX6, P21, et al). Finally, the ANKRD11 KD-mediated influence on cell proliferation and differentiation were reversed by inhibiting histone acetyltransferase activity. Conclusion Our assay defined that ANKRD11 was a crucial chromatin regulator that suppress histone acetylation and then decrease gene expression during myeloid differentiation, providing a likely explanation for its role in MDS pathogenesis. This study further support histone acetylase inhibitor as a potential treatment in MDS. Figure ANKRD11mutation distribution (a) and coexist with other mutations (b). Figure. ANKRD11mutation distribution (a) and coexist with other mutations (b). Figure The mRNA expression levels of ANKRD11in our MDS (A, C) subset and GEO data (B). Figure. The mRNA expression levels of ANKRD11in our MDS (A, C) subset and GEO data (B). Changes of histone acetylation in ANKRD11-KD cell line (MEG-01). ANKRD11 KD significantly increased acetylation of histones H3 and H4 at H3K14 and H4K5. Changes of histone acetylation in ANKRD11-KD cell line (MEG-01). ANKRD11 KD significantly increased acetylation of histones H3 and H4 at H3K14 and H4K5. Disclosures No relevant conflicts of interest to declare.

scholarly journals Furanoic Lipid F-6, A Novel Anti-Cancer Compound that Kills Cancer Cells by Suppressing Proliferation and Inducing Apoptosis

Cancers ◽  
2019 ◽  
Vol 11 (7) ◽  
pp. 960 ◽  
Author(s):  
Jassim M. Al-Hassan ◽  
Yuan Fang Liu ◽  
Meraj A. Khan ◽  
Peiying Yang ◽  
Rui Guan ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Cancer Cells ◽  
Cell Lines ◽  
Cancer Cell ◽  
Neutrophil Extracellular Trap ◽  
Cancer Drug ◽  
Drug Candidate ◽  
Cell Recovery ◽  
Network Analyses ◽  
Anti Cancer

Identifying novel anti-cancer drugs is important for devising better cancer treatment options. In a series of studies designed to identify novel therapeutic compounds, we recently showed that a C-20 fatty acid (12,15-epoxy-13,14-dimethyleicosa-12,14-dienoic acid, a furanoic acid or F-6) present in the lipid fraction of the secretions of the Arabian Gulf catfish skin (Arius bilineatus Val.; AGCS) robustly induces neutrophil extracellular trap formation. Here, we demonstrate that a lipid mix (Ft-3) extracted from AGCS and F-6, a component of Ft-3, dose dependently kill two cancer cell lines (leukemic K-562 and breast MDA MB-231). Pure F-6 is approximately 3.5 to 16 times more effective than Ft-3 in killing these cancer cells, respectively. Multiplex assays and network analyses show that F-6 promotes the activation of MAPKs such as Erk, JNK, and p38, and specifically suppresses JNK-mediated c-Jun activation necessary for AP-1-mediated cell survival pathways. In both cell lines, F-6 suppresses PI3K-Akt-mTOR pathway specific proteins, indicating that cell proliferation and Akt-mediated protection of mitochondrial stability are compromised by this treatment. Western blot analyses of cleaved caspase 3 (cCasp3) and poly ADP ribose polymerase (PARP) confirmed that F-6 dose-dependently induced apoptosis in both of these cell lines. In 14-day cell recovery experiments, cells treated with increasing doses of F-6 and Ft-3 fail to recover after subsequent drug washout. In summary, this study demonstrates that C-20 furanoic acid F-6, suppresses cancer cell proliferation and promotes apoptotic cell death in leukemic and breast cancer cells, and prevents cell recovery. Therefore, F-6 is a potential anti-cancer drug candidate.

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