scholarly journals Potential Metabolite Nymphayol Isolated from Water Lily (Nymphaea stellata) Flower Inhibits MCF-7 Human Breast Cancer Cell Growth via Upregulation of Cdkn2a, pRb2, p53 and Downregulation of PCNA mRNA Expressions

Metabolites ◽  
2020 ◽  
Vol 10 (7) ◽  
pp. 280
Author(s):  
Laila Naif Al-Harbi ◽  
Pandurangan Subash-Babu ◽  
Manal Abdulaziz Binobead ◽  
Maha Hussain Alhussain ◽  
Sahar Abdulaziz AlSedairy ◽  
...  

Controlled production of cyclin dependent kinases (CDK) and stabilization of tumor suppressor genes are the most important factors involved in preventing carcinogenesis. The present study aimed to explore the cyclin dependent apoptotic effect of nymphayol on breast cancer MCF-7 cells. In our previous study, we isolated the crystal from a chloroform extract of Nymphaea stellata flower petals and it was confirmed as nymphayol (17-(hexan-2-yl)-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-3-ol) using x-ray diffraction (XRD), Fourier transform infrared (FTIR), and mass spectroscopy (MS) methods. The cytotoxic effect of nymphayol on MCF-7 cells were analyzed using the 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. The cellular and nuclear damage was determined using propidium iodide (PI) and acridine orange/ethidium bromide (AO/ErBr) staining. Tumor suppressor and apoptosis related mRNA transcript levels were determined using real-time polymerase chain reaction (RT-PCR). Nymphayol potentially inhibits MCF-7 cell viability up to 78%, and the IC50 value was observed as 2.8 µM in 24 h and 1.4 µM in 48 h. Treatment with nymphayol significantly increased reactive oxygen species (ROS) level and the tunnel assay confirmed DNA damage. We found characteristically 76% apoptotic cells and 9% necrotic cells in PI and AO/ErBr staining after 48 h treatment with 2.8 µM of nymphayol. Gene expression analysis confirmed significantly (p ≤ 0.001) increased mRNA levels of cyclin dependent kinase inhibitor 2A (Cdkn2a), retinoblastoma protein 2 (pRb2), p53, nuclear factor erythroid 2-factor 2 (Nrf2), caspase-3, and decreased B-cell lymphoma 2 (Bcl-2), murine double minute 2 (mdm2), and proliferating cell nuclear antigen (PCNA) expression after 48 h. Nymphayol effectively inhibited breast cancer cell viability, and is associated with early expression of Cdkn2a, pRb2, and activation of p53 and caspases.

2018 ◽  
Vol 10 (1) ◽  
pp. 335
Author(s):  
Sandy Vitria Kurniawan ◽  
Lies Sugiarti ◽  
Septelia Inawati Wanandi ◽  
Melva Louisa

Objective: This study was designed to analyze the role of piperine in modulating P-glycoprotein mRNA expression when added in combination withtamoxifen to breast cancer cells in culture.Methods: MCF-7 breast cancer cells were treated with 1 μM tamoxifen with or without piperine (12.5, 25, or 50 μM) or verapamil 50 μM (P-glycoproteininhibitor positive control) for up to 12 days. We assessed the cell viability and isolated total RNA from them. We quantified P-glycoprotein expressionsusing quantitative reverse transcription polymerase chain reaction.Results: Administration of various doses of piperine decreased MCF-7 breast cancer cell viability. Piperine, when given in combination with tamoxifen,decreased the expression of P-glycoprotein mRNA in cells compared with the expression in cells treated with tamoxifen only. The effects were shownto be dose dependent.Conclusion: Piperine prevents the development of breast cancer cell tamoxifen resistance, probably through its inhibition of P-glycoprotein expression.


1997 ◽  
Vol 82 (6) ◽  
pp. 1790-1798 ◽  
Author(s):  
Rama Natarajan ◽  
Robert Esworthy ◽  
Wei Bai ◽  
Jia-Li Gu ◽  
Sharon Wilczynski ◽  
...  

Abstract The interaction of growth factors, such as epidermal growth factor (EGF) with their receptors, on breast cancer cells can lead to the hydrolysis of phospholipids and release of fatty acids, such as arachidonic acid, which can be further metabolized by the lipoxygenase (LO) pathway. Several LO products have been shown to stimulate oncogenes and have mitogenic and chemotactic effects. In this study, we have evaluated the regulation of 12-LO activity and expression in breast cancer cells and tissues. Leukocyte-type 12-LO messenger RNA (mRNA) expression was studied by a specific RT-PCR method in matched, normal, uninvolved and cancer-involved breast tissue RNA samples from six patients. In each of these six patients, the cancer-involved section showed a much higher level of 12-LO mRNA than the corresponding normal section. 12-LO mRNA levels also were greater in two breast cancer cell lines, MCF-7 and COH-BR1, compared with the nontumorigenic breast epithelial cell line, MCF-10F. The growth of the MCF-7 cells was significantly inhibited by two specific LO blockers but not by a cyclooxygenase blocker. Treatment of serum-starved MCF-7 cells with EGF for 4 h led to a dose-dependent increase in the formation of the 12-LO product, 12-hydroxyeicosatetraenoic acid. EGF treatment also increased the levels of the leukocyte-type 12-LO protein expression at 24 h. These results suggest that activation of the 12-LO pathway may play a key role in basal and EGF-induced breast cancer cell growth.


2022 ◽  
Vol 12 (5) ◽  
pp. 989-995
Author(s):  
Ke Chunlin ◽  
Dong Feng ◽  
Wang Peirong

Objective: The purpose of our study was to evaluate Enhancement Mechanism of Dihydromyricetin (DMY) on the Inhibitory Role of Cisplatin Towards Breast Cancer Cell Activity. Materials and Methods: The MCF-7 were divided into NC, DMY, Cis and DMY+Cis groups. Using relative methods (MTT, TUNEL, Transwell, flow cytometry and wound healing) to evaluate MCF-7 cell biological activities including cell viability, apoptosis, invasion cell number and wound healing rate. The relative proteins expressions including FOXO-1, Noxa, Bim, Cyto C, Caspase-3, Caspase-9 and Apaf-1 were evaluated by WB assay. Results: MCF-7 cell viability, invasion cell number and wound healing rates were significantly depressed and apoptosis rate were significantly increased in DMY, Cis and DMY+Cis groups (P < 0.01, respectively). Compared with Cis group, cell viability, invasion cell number and wound healing rates were significantly depressed and apoptosis rate were significantly increased in DMY+Cis group (P < 0.05, respectively). Conclusion: Dihydromyricetin can effectively enhance the inhibitory effect of cisplatin on breast cancer cells.


2017 ◽  
Vol 95 (4) ◽  
pp. 468-473 ◽  
Author(s):  
Seyedeh Tayebeh Rahideh ◽  
Mohammad Keramatipour ◽  
Mitra Nourbakhsh ◽  
Fariba Koohdani ◽  
Mostafa Hoseini ◽  
...  

Nobiletin (NOB) is one of the polymethoxyflavones mainly found in citrus fruits. Aromatase or cytochrome P450 (CYP19) enzyme catalyzes the last and rate-limiting step in estrogen biosynthesis. This study was carried out to investigate the effect of NOB on the activity and expression of aromatase, and to compare this property with letrozole (LET) as aromatase inhibitor in the MCF-7 breast cancer cell line. Cell viability was assessed with 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) assays. Aromatase enzyme activity based on the conversion of androgenic substrate testosterone into 17β-estradiol was determined. CYP19 gene expression was measured by quantitative real-time PCR. MTT assays demonstrated that NOB at a concentration of 100 μmol/L decreased cell viability in a time-dependent manner (P < 0.05). NOB significantly inhibited aromatase at the concentration of 0.1 μmol/L (P = 0.013), whereas other concentrations had no effect. Treatment with 10 μmol/L and 1 μmol/L of NOB for 48 h significantly increased (P = 0.001) and decreased (P = 0.02) relative aromatase expression, respectively. The combination of LET and NOB had no effect on aromatase. This study showed for the first time that NOB decreases the activity and expression of aromatase at low concentrations in MCF-7 breast cancer cells.


2020 ◽  
Author(s):  
Marzieh Marzbany ◽  
Amir Hossein Norooznezhad ◽  
Zohreh Hoseinkhani ◽  
Azadeh Mahnam ◽  
Kiumaras Eslampia ◽  
...  

Abstract Background: Testis-specific gene antigen (TSGA10) mainly involves in spermatogenesis and embryogenesis. In the new defined roles, being a tumor suppressor agent or a cancer/testis antigen (CTA) is still unclear for this protein. The current study aimed to examine exact role of TSGA10 as a tumor suppressor or CTA in breast cancer and evaluate the role of microenvironment on its expression. Methods: This study evaluated the expression of TSGA10 and hypoxia-inducible factor 1α (HIF-1α) in two different breast cancer cell lines (MCF-7 and MDA-MB23) as well as their control (MCF10A) using real-time PCR. Moreover, expression of the mentioned genes evaluated in samples obtained from tumoral tissues with two types of controls: paired (tumor-free margin) and unpaired (healthy individuals). Also, in order to asses TSGA10 levels in the tumoral tissues, western blotting was performed. Furthermore, to evaluate the role epigenetic changes on TSGA10 expression, breast cancer cell lines were treated with a histone deacetylase inhibitor (HDACI) as well as H2O2 for oxidative stress induction. Results: The current study evaluated 36 patients diagnosed with breast cancer as well as 10 healthy controls. According to the results, it was shown that 35 (97.7%) and 1 (2.8%) of patients were diagnosed with ductal and lobular carcinomas respectively. The TSGA10 levels in the tumoral samples showed 1.38±0.014-fold decrease and 1.41±0.127-fold increase compared with their paired (P<0.001) and unpaired (P<0.001) controls respectively. Moreover, results of blotting in tumoral tissues expressed significant decrease in TSGA10 levels in comparison to the paired controls (P<0.01). Among the cell lines, TSGA10 expression in MCF-7 and MDA-MB23 cells had 4.9±0.283 and 4.21±0.163 folds of decrease in normoxic and 4.7±.0.283 and 7.1±0.141 folds of expression reduction in hypoxic condition respectively (all P<0.0001). Furthermore, the results showed that HIF-1α expression was up-regulated in both normoxic (P<0.01) and hypoxic (P<0.01) conditions. Also, TSGA10 expression increased up to 7.39±0.156 folds in MCF-7 cells after HDACI treatment (all P<0.01). However, MDA-MB23 cells firstly experienced a decrease and then a notable increase in TSGA10 expression (all P<0.01). Conclusion: Results of current study showed that TSGA10 seems to be tumor suppressor, however, further studies are necessary.


Author(s):  
Abd Elmoneim O. Elkhalifa ◽  
Eyad Al-Shammari ◽  
Mohammad Jahoor Alam ◽  
Jerold C. Alcantara ◽  
Mushtaq Ahmad Khan ◽  
...  

Objective: Cancer chemoprevention with phytochemicals such as “lutein” derived from the vegetable okra could prove beneficial. Therefore, the objective of this study was to perform a meta-analysis of “lutein” against the breast cancer cell lines (MCF-7) and to establish the possible development of lutein based nutraceuticals. Methodology: A literature survey was performed using online data bases such as PubMed, Google scholar, and EMBASE, from 2000 to 2020 by using keywords such as “Lutein”, “Anticancer activity”, “Breast cancer cell lines”, and “MCF-7”. Studies reported lutein anticancer potentials against MCF-7 were included in the study. Results: Out of 28 studies, 7 research articles fulfilled the inclusion criteria. Meta-analysis data indicated that, a lutein concentration at ≥1 µM was able to reduce the MCF-7 cell viability of 59.837 with a 95% confidence interval (CI): 48.331 to 71.343. Additionally, a forest plot of the cumulative studies also indicated that impact of lutein concentration to reduce the MCF-7 cell viability was around 60%. Moreover, the I2 value of lutein was 74%, which is a considerable heterogeneity. Conclusion: Therefore, based upon the meta-analysis data, the conclusion is that dietary lutein supplementation and fortification of food with clinical data could be an approach to develop a nutraceutical product for preventive, as well as for adjunct therapeutic purposes in various breast cancer subtypes. 


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