scholarly journals Evaluation of Ginkgo biloba and Flunixin in BIM Gene Expression and Viability of Ovarian Cancer Cell A2780s

Author(s):  
Farnoosh Soleamani ◽  
Elham Salehi ◽  
Majid Morovati-Sharifabad ◽  
Fatemeh Sarkargar ◽  
Gholamhosein Pourghanbari

Background: Ovarian cancer is the deadliest gynecologic cancer. Studies on the therapeutic properties of Ginkgo biloba and flunixin showed that these drugs, singly or in combination with other drugs, have anti-cancer activities. Different genes are involved in apoptosis regulation. The BIM gene is one of the most important regulators of this process. BIM has different roles, including cell cycle regulation, apoptosis induction, deoxyribonucleic acid recombination, chromosomal segregation, and cell aging. Methods: This study evaluated the viability percentage of the A2780s cell line with Ginkgo biloba and flunixin at different concentrations, compared to that of the control group. Then, the half-maximal inhibitory concentration (IC50) values of Ginkgo biloba and flunixin were determined within 24 h. Then, the expression of the BIM gene was evaluated using a real-time polymerase chain reaction (PCR). Results: The IC50 results showed that Ginkgo biloba and flunixin significantly reduced cell life (P < 0.01) depending on time and concentration. The results of real-time PCR showed that cell treatment with Ginkgo biloba and flunixin significantly increased BIM expression. Conclusions: The results of this experiment indicated that BIM gene expression was increased in cancer cells treated with Ginkgo biloba and flunixin, compared to that reported for control cells. Therefore, with further research in the future, these compounds can be used for the development of ovarian anti-cancer drugs.

2021 ◽  
Vol In Press (In Press) ◽  
Author(s):  
Maryam Yahyaie ◽  
Majid Morovati-Sharifabad ◽  
Elham Salehi ◽  
Fatemeh Sarkargar ◽  
Gholamhosein Pourghanbari

Background: Ovarian cancer is the second leading cause of death in Iran compared with other gynecological diseases. Considering the role of cyclooxygenase (COX) enzymes and prostaglandin E2 production in tumor lesions, nonsteroidal anti-inflammatory drugs (NSAIDs) show antitumor properties by inhibiting COX. Furthermore, some compounds can serve as non-selective inhibitors of COX (such as ketoprofen) and prevent cancer development. Human epididymis protein 4 (HE4) is one of the most sensitive tumor markers known in the study of the disease of ovarian epithelial cancer. The expression of HE4 increases in different types of ovarian cancer. Objectives: The aim of this study was to determine the anti-cancer effects of ketoprofen on the viability of ovarian cancer cells and expression of HE4 gene. Methods: To calculate half-maximal inhibitory concentration (IC50), A2780S cells were treated with different concentrations of ketoprofen for 24 hours, then the cells were incubated with appropriate concentrations of IC50 for 24, 48, and 72 hours. Real-time polymerase chain reaction (PCR) was used to measure changes caused by the effect of drugs on HE4 gene expression and analyzed by the 2-ΔΔCT method. Results: The IC50 level of ketoprofen for 24 hours was 583.7 μM. According to real-time PCR results, treatment of cells with ketoprofen reduced HE4 expression. Conclusions: HE4 gene expression decreased in cells treated with ketoprofen compared with the cells in the control group, which proves the anti-cancer activity of ketoprofen and a reduction in the viability of ovarian cancer cells.


2020 ◽  
Author(s):  
Shahan Mamoor

Ovarian cancer is the most lethal gynecologic cancer (1). We sought to identify genes associated with high-grade serous ovarian cancer (HGSC) by comparing global gene expression profiles of normal ovary with that of primary tumors from women diagnosed with HGSC using published microarray data (2, 3). We identified the forkhead box L2, (FOXL2) (4) as among the genes whose expression was most different in HGSC ovarian tumors. FOXL2 expression was significantly lower in ovarian tumors relative to normal ovary. FOXL2 has established roles in ovarian development (4, 5), and the FOXL2 gene is mutated in granulosa-cell tumors of the ovary (6). These data indicate FOXL2 might also be perturbed, at the level of gene expression, in high-grade serous ovarian cancers.


2021 ◽  
Author(s):  
Courtney Griffiths ◽  
Michelle Bilbao ◽  
Lauren Krill ◽  
Olga Ostrovsky

Early diagnosis and intervention are some of the longstanding challenges associated with ovarian cancer, which is the leading cause of gynecologic cancer mortality. While the majority of patients who present with advanced stage disease at time of diagnosis will initially respond to traditional combination platinum and taxane-based chemotherapy in conjunction with cytoreductive surgery, approximately 70% will ultimately recur due to chemoresistance within the first two years. Intratumor heterogeneity is proposed to be a leading factor in the development of chemoresistance and resultant poorer outcomes for those with recurrent or advanced stage disease. Both inherent and acquired mechanisms of chemoresistance are postulated to be a result of alterations in gene expression, also known as epigenetic modifications. Therefore, epigenetic therapy is a pivotal avenue which allows for reversal of chemoresistance in cancer through the targeting of aberrant mutations. In this chapter, we discuss how these epigenetic modifications prove to be promising targets in cancer therapy leading to heightened drug sensitivity and improved patient survival outcomes.


Author(s):  
I. Ilienko ◽  
◽  
D. Bazyka ◽  
N. Golyarnyk ◽  
L. Zvarych ◽  
...  

Objective. to establish the connection of radiation-induced changes in gene expression with the realized pathology of the broncho-pulmonary and cardiovascular systems in Chornobyl clean-up workers. Materials and methods. We examined 314 male Chornobyl clean-up workers (main group; age (58.94 ± 6.82) years (M ± SD); min 33, max 79 years; radiation dose (411.82 ± 625.41) mSv (M ± SD); min 1.74, max 3600 mSv) with various nosological forms of cardiovascular and broncho-pulmonary pathology (BPP) and 50 subjects of the control group: age (50.50 ± 5.73) years (M ± SD); min 41, max 67 years. The relative level of BCL2, CDKN2A, CLSTN2, GSTM1, IFNG, IL1B, MCF2L, SERPINB9, STAT3, TERF1, TERF2, TERT, TNF, TP53, CCND1, CSF2, VEGFA genes expression was determined in peripheral blood leukocytes by real-time PCR (7900 HT Fast Real-Time PCR System (Applied Biosystems, USA)). The «gene-disease» association was determined on statistical models stratified separately for each disease and gene. Logistic regression was used to calculate the odds ratio. Results. Increased GSTM1 gene expression and no changes in angiogenesis-related VEGFA gene expression were found in the main group of patients with coronary heart disease (CHD). It was established overexpression of TP53, VEGF and IFNG genes in the group of patients with arterial hypertension (AH). At combination of these diseases an increase of expression of СSF2, TERF1, TERF2 genes was established. The detected changes demonstrate an activation of the antioxidative defense system in patients with CHD, while AH is associated with the expression of genes of angiogenesis and immune inflammation. It was shown an increase in the expression of genes associated with apoptosis and kinase activity (BCL2, CLSTN2, CDKN2), immune inflammation (CSF2, IL1B, TNF) in Chornobyl clean-up workers with BPP. Expression of TP53 and GSTM1 (gene, associated with the glutathione system) was significantly upregulated in the group of individuals with chronic bronchitis, whereas in patients with chronic obstructive pulmonary disease, no increase was detected; the expression of SERPINB9 and MCF2L genes was downregulated. Conclusions. Changes in the expression of genes, associated with the development of somatic pathology in the remote period after irradiation, in particular the genes of the immune response and inflammatory reactions CSF2, IFNG, IL1B, TNF; expression of genes that regulate cell proliferation, aging and apoptosis TP53, BCL2, MCF2L, CDKN2A, SERPINB9, TERF1, TERF2, TERT; genes that regulate cell adhesion and angiogenesis CLSTN2, VEGF. Key words: gene expression, somatic pathology, radiation, Chornobyl.


Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 4088-4088
Author(s):  
Weidong Lu ◽  
Ursula A. Metulonis ◽  
Anne Doherty-Gilman ◽  
Hang Lee ◽  
Elizabeth Dean-Clower ◽  
...  

Abstract Purpose Ovarian cancer is the most lethal gynecologic cancer. Chemotherapy, the standard of care, has hematologic toxicity, primarily neutropenia. G-CSF is currently used to support white blood cell (WBC) and absolute neutrophil counts (ANC). Prior clinical trials from China suggest that acupuncture could ameliorate chemotherapy-induced leukopenia; the proposed mechanism is an increase in G-CSF levels. In the current study, we investigated the effect of acupuncture, administered during myelosuppressive therapy, on WBC and ANC counts in ovarian cancer patients. Patients and methods Twenty-one newly diagnosed or recurrent ovarian cancer patients were randomized to receive active versus sham acupuncture while undergoing standard IV platinum and taxane-containing chemotherapy. A standardized protocol with 9 acupuncture points was employed with manual and electroacupuncture stimulation. The frequency of acupuncture treatment was 2–3 times per week for a total of 10 sessions, starting 1 week before the 2nd cycle of chemotherapy. WBC and ANC counts were checked weekly at five time points. Serum G-CSF was collected four times during the study. Results Of 587 patients screened, 21 patients were enrolled and received either acupuncture or sham treatment. Patients in both the active and control arms had similar patient characteristics and treatment. Both median WBC and ANC values at nadir in the acupuncture arm were higher than in the control arm, but the differences were not statistically significant, after adjusting for the baseline difference. However, the median WBC in the acupuncture arm at recovery was statistically significantly higher than the control arm, after adjustment (8,600 cell/μL, range: 4,800–12,000 vs. 4,400 cell/μL range: 2,300–10,000) (p=0.045). The recovering median ANC in the patients receiving acupuncture also was higher, but this difference was not statistically significant (p=0.094). The median serum G-CSF at baseline for patients in the active vs. control arm was similar (37.3 pg/mL, range 28.6–393.3 vs. 32.0, range 11.8–211.3, respectively) (p=0.291). At the second time point, the 1st day of the 2nd cycle, the acupuncture group had a higher G-CSF value than the control group (p=0.121). At nadir, the acupuncture group still had a slightly higher G-CSF value than in the control group (p=0.796). However, at the recovery day, the 1st day of 3rd cycle, the G-CSF value in the acupuncture group was lower than in the control arm (p=0.729). No statistical significance in G-CSF value was found at each time point between the two groups. Conclusion The acupuncture protocol used in this study was feasible and safe. We report trends of higher WBC and ANC values during one cycle of myelosuppressive chemotherapy in ovarian cancer patients, suggesting a potential myeloprotective effect of acupuncture. However, current data do not support an acupuncture effect on G-CSF production. These findings warrant a larger study to explore the observed clinical trends and other potential underlying mechanisms.


2019 ◽  
Vol 19 (6) ◽  
pp. 820-826 ◽  
Author(s):  
Mohsen Rashid ◽  
Forough Sanjarin ◽  
Farzaneh Sabouni

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.


Author(s):  
Zeinolabedin Sharifian ◽  
Batool Hashemibeni ◽  
Majid Pourentezari ◽  
Ali Valiani ◽  
Mohammad Mardani ◽  
...  

Background and Aims: Tissue engineering is a relatively novel field that has been intensely developing during recent years and has shown to be excessively promising when used for cartilage regeneration. Scaffolds represent important components for tissue engineering. Materials and Methods: The Poly Lactic-Co-Glycolic Acid (PLGA) impregnated with fibrin and hyaluronic acid (HA) produce hybrid scaffolds. human adipose-derived stem cells (hADSCs) were seeded in scaffolds and cultured in chondrogenic media. The viability of cells in different groups was assessed by MTT. The expression of chondrogenic related genes [Sox9, type II collagen (Col II), Aggrecan(AGG)] and type X collagen (Col X) was quantified by real-time polymerase chain reaction. Results: The results of the real-time PCR showed SOX9, AGG and Col X gene expression in the control groups being significantly lower than the other groups (p≤0.05). It also demonstrated Col II gene expression in the control group being significantly lower than the PLGA/Fibrin and PLGA/Fibrin/HA groups (p≤0.05). The Col X gene expression of cells in PLGA/HA and PLGA/Fibrin/HA groups significantly decreased in comparison with the PLGA/Fibrin group (p≤0.05). Conclusions: These conclusions indicate that administration of PLGA/ Fibrin and PLGA/HA scaffolds, particularly PLGA/Fibrin/ HA, motivates chondrogenesis in hADSCs. This can be diminished by decreasing hypertrophic markers and increasing characteristic markers of hyaline cartilage.


Drug Research ◽  
2017 ◽  
Vol 68 (04) ◽  
pp. 213-221 ◽  
Author(s):  
Samaneh Pirmoradi ◽  
Ezzatollah Fathi ◽  
Raheleh Farahzadi ◽  
Younes Pilehvar-Soltanahmadi ◽  
Nosratollah Zarghami

AbstractAging and losing cell survival is one of the main problems in cell therapy. Aging of Mesenchymal Stem Cells (MSCs) is associated with a rise in intracellular reactive oxygen species, decrease in telomerase reverse transcriptase (TERT) expression and finally eroded telomere ends. Given that the production of free radicals in the aging process is effective, the use of antioxidants can help in scavenging free radicals and prevent the aging of cells. The aim of this study is to evaluate the effects of curcumin on proliferation, aging and TERT expression of rat adipose tissue-derived stem cells (rADSC). rADSCs were isolated from inguinal rat adipose tissue and their viabilities were assessed by MTT after exposure to different concentrations of curcumin. Flow-cytometry was performed for investigating the cell surface markers. Adipogenic and osteogenic differentiation were carried out to evaluate the pluripotency of rADSCs. Telomerase expression and percentage of senescent cells were evaluated using real-time PCR and senescence-associated β-galactosidase activity, respectively. The results demonstrated significant proliferation of rADSCs after 48 h treatment with 1 and 5 µM curcumin. Additionally, these concentrations could significantly reduce the population doubling time and aging of rADSCs at different passages. The findings of SA-ß-gal staining showed that curcumin significantly decreased the number of senescent cells in the 5 and 7 cell passages. Moreover, expression levels of TERT increased in the presence of 1 and 5 µM curcumin than control group (P<0.001). As a conclusion, curcumin may be a good candidate to improve lifespan of rADSCs through promoting TERT gene expression.


2018 ◽  
Vol 5 (4) ◽  
pp. 31 ◽  
Author(s):  
Maryam Samareh Salavati Pour ◽  
Fatemeh Hoseinpour Kasgari ◽  
Alireza Farsinejad ◽  
Ahmad Fatemi ◽  
Roohollah Mirzaee Khalilabadi

Introduction: Mesenchymal stem cells (MSCs) are widely studied due to their self- renewal potential and capacity to differentiate into multiple tissues. However, they have a limited life span of several divisions in vitro, which alters various cellular characteristics and reduces their application. Aim: We evaluated the effect of platelet-derived microparticles on gene expression of hTERT, one of the main factors involved in aging and cell longevity. Materials and methods: Umbilical cord MSCs were used for this study. Cells were characterized by evaluating morphology via inverted microscope and identifying associated surface markers using flow cytometry. Platelet-derived microparticles were prepared by centrifuging platelet bags at varying speeds, and their concen- trations were determined by Bradford assay. At 30% confluency, MSCs were treated with 50 μg/mL of microparticles for five days. Then, RNA was extracted and cDNA was synthesized. Quantitative expression of hTERT was assessed using real-time polymerase chain reaction (PCR). Results: Fibroblast-like cells were isolated from umbilical cord tissue and MSCs were identified by the presence of mesenchymal surface markers via flow cytometry. Real- time PCR showed that gene expression of hTERT increased by more than three times when treated with platelet-derived microparticles, in comparison to expression of the control group. Conclusion: We concluded that platelet-derived microparticles may be a potentially safe and effective method to increase hTERT gene expression in MSCs, ultimately prolonging their life span in vitro. 


Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 5212-5212
Author(s):  
Panagiotis Theodorou Diamantopoulos ◽  
Katerina Polonyfi ◽  
Nikolaos Spanakis ◽  
Athanassios Galanopoulos ◽  
Georgia Diamantopoulou ◽  
...  

Abstract Abstract 5212 Background: In all Epstein-Barr (EBV)-associated malignancies, the virus displays a latency program of infection and a restricted pattern of gene expression. Among the products of these genes, latent membrane protein 1 (LMP1) is a potent transforming protein with several different roles. LMP1 has been shown in cell lines to stimulate apoptosis. Survivin, a member of the inhibitor of apoptosis (IAP) family, is an important regulator of the mitochondrial apoptotic pathway, while oxidative stress (OS) is a cellular condition particularly relevant to cell aging. In the present study we enrolled patients with non-EBV-related low grade B-cell lymphoproliferative diseases. The aim was to detect (1) the viral load of EBV-positive patients, (2) the expression of LMP1 oncoprotein, (3) the possible apoptotic properties of LMP1 by correlating the levels of survivin with LMP1 expression, and (4) the levels of oxidative stress in LMP1-positive and negative patients. Patients and Methods: Forty eight Greek patients with EBV-unrelated low grade B-cell leukemic lymphomas, were enrolled in the study (chronic lymphocytic leukemia: 27, marginal zone lymphoma: 12, mantle cell lymphoma: 4, hairy cell leukemia: 2, follicular lymphoma: 2, lymphoplasmacytic lymphoma: 1). The majority of patients (61.2%) were treatment-naïve, while the rest had not received any treatment for at least 6 months. DNA from peripheral blood was tested by quantitative real time (qRT) PCR for the EBV-R gene. RNA from EBV-positive patients was examined by RT-PCR and qRT PCR for LMP-1, while using qRT PCR we measured survivin expression in all patients. Densitometric analysis (DA) was used for semi-quantification of the survivin gene expression. The results were expressed relative to the expression of ABL housekeeping gene. The control group included 30 EBV-negative healthy adults. Oxidative stress was measured in the serum of all patients using the PerOx (TOS/TOC) Kit, by Immunodiagnostik. Non parametric methods (Mann-Whitney test) were used for statistical analysis of the results. Results: Twenty five (25) men and 23 women, with a median age of 74 (51–87 years old) were studied. EBV positivity was detected in 19/48 (39.6%) patients, and LMP1 was expressed in 13/19 (68.4%) EBV-positive patients. Survivin levels were lower in LMP1-positive patients vs LMP1-negative patients (2-tailed p=0.009). The oxidative stress was lower (261.4 μmol/L) in LMP1-positive patients vs LMP1-negative patients (372.3 μmol/L), (2-tailed p=0.014). Discussion: The literature lacks information about the expression of LMP1 in the peripheral blood of patients with non-EBV-related low grade B-cell leukemic lymphomas. Previous studies in LMP1-positive lymphoma cell lines have shown the apoptotic functions of LMP1 during type II latency. In this study LMP1-positive patients express statistically significant lower levels of survivin vs LMP1-negative patients. This finding is in accordance to the hypothesis that LMP1 oncoprotein can induce apoptosis. LMP1-positive patients had lower levels of oxidative stress compared to LMP1-negative patients. According to our findings, in non-EBV-related lymphomas, LMP1 may increase apoptosis and decrease the levels of oxidative stress. Disclosures: No relevant conflicts of interest to declare.


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