scholarly journals MCF-7 Drug Resistant Cell Lines Switch Their Lipid Metabolism to Triple Negative Breast Cancer Signature

Cancers ◽  
2021 ◽  
Vol 13 (23) ◽  
pp. 5871
Author(s):  
Jose Adriá-Cebrián ◽  
Sandra Guaita-Esteruelas ◽  
Eric W.-F. Lam ◽  
Marta Rodríguez-Balada ◽  
Jordi Capellades ◽  
...  

Obesity and adipose tissue have been closely related to a poor cancer prognosis, especially in prostate and breast cancer patients. The ability of transferring lipids from the adipose tissue to the tumor cells is actively linked to tumor progression. However, different types of breast tumor seem to use these lipids in different ways and metabolize them in different pathways. In this study we have tracked by mass spectrometry how palmitic acid from the adipocytes is released to media being later incorporated in different breast cancer cell lines (MDA-MB-231, SKBR3, BT474, MCF-7 and its resistant MCF-7 EPIR and MCF-7 TAXR). We have observed that different lines metabolize the palmitic acid in a different way and use their carbons in the synthesis of different new lipid families. Furthermore, we have observed that the lipid synthesis pattern varied according to the cell line. Surprisingly, the metabolic pattern of the resistant cells was more related to the TNBC cell line compared to their sensitive cell line MCF-7. These results allow us to determine a specific lipid pattern in different cell lines that later might be used in breast cancer diagnosis and to find a better treatment according to the cancer molecular type.

2020 ◽  
Author(s):  
Adriane Feijo Evangelista ◽  
Renato J Oliveira ◽  
Viviane A O Silva ◽  
Rene A D C Vieira ◽  
Rui M Reis ◽  
...  

Abstract Introduction: Breast cancer is the most frequently diagnosed malignancy among women. However, the role of microRNA expression in breast cancer progression is not fully understood. In this study we examined predictive interactions between differentially expressed miRNAs and mRNAs in breast cancer cell lines representative of the common molecular subtypes. Integrative bioinformatics analysis identified miR-193 and miR-210 as potential regulatory biomarkers of mRNA in breast cancer. Several recent studies have investigated these miRNAs in a broad range of tumors, but the mechanism of their involvement in cancer progression has not previously been investigated. Methods: The miRNA-mRNA interactions in breast cancer cell lines were identified by parallel expression analysis and miRNA target prediction programs. The expression profiles of mRNA and miRNAs from luminal (MCF-7, MCF-7/AZ and T47D), HER2 (BT20 and SK-BR3) and triple negative subtypes (Hs578T e MDA-MB-231) could be clearly separated by unsupervised analysis using HB4A cell line as a control. Breast cancer miRNA data from TCGA patients were grouped according to molecular subtypes and then used to validate these findings. Expression of miR-193 and miR-210 was investigated by miRNA transient silencing assays using the MCF7, BT20 and MDA-MB-231 cell lines. Functional studies included, xCELLigence system, ApoTox-Glo triplex, flow cytometry and transwell assays were performed to determine cell proliferation, cytotoxicity, apoptosis, migration and invasion, respectively. Results: The most evident effects were associated with cell proliferation after miR-210 silencing in triple negative subtype cell line MDA-MB-231. Using in silico prediction algorithms, TNFRSF10 was identified as one of the potential downstream targets for both miRNAs. The TNFRSF10C and TNFRSF10D mRNA expression inversely correlated with the expression levels of miR-193 and miR210 in breast cell lines and breast cancer patients, respectively. Other potential regulated genes whose expression also inversely correlated with both miRNAs were CCND1, a mediator on invasion and metastasis, and the tumor suppressor gene RUNX3. Conclusion: In summary, our findings identify miR-193 and miR-210 as potential regulatory miRNA in different molecular subtypes of breast cancer and suggest that miR-210 may have specific role in MDA-MB-231 proliferation. Our results highlight important new downstream regulated targets that may serve as promising therapeutic pathways for aggressive breast cancers.


2021 ◽  
Author(s):  
Adriane Feijo Evangelista ◽  
Renato J Oliveira ◽  
Viviane A O Silva ◽  
Rene A D C Vieira ◽  
Rui M Reis ◽  
...  

Abstract Background: Breast cancer is the most frequently diagnosed malignancy among women. However, the role of microRNA (miRNA) expression in breast cancer progression is not fully understood. In this study we examined predictive interactions between differentially expressed miRNAs and mRNAs in breast cancer cell lines representative of the common molecular subtypes. Integrative bioinformatics analysis identified miR-193 and miR-210 as potential regulatory biomarkers of mRNA in breast cancer. Several recent studies have investigated these miRNAs in a broad range of tumors, but the mechanism of their involvement in cancer progression has not previously been investigated.Methods: The miRNA-mRNA interactions in breast cancer cell lines were identified by parallel expression analysis and miRNA target prediction programs. The expression profiles of mRNA and miRNAs from luminal (MCF-7, MCF-7/AZ and T47D), HER2 (BT20 and SK-BR3) and triple negative subtypes (Hs578T e MDA-MB-231) could be clearly separated by unsupervised analysis using HB4A cell line as a control. Breast cancer miRNA data from TCGA patients were grouped according to molecular subtypes and then used to validate these findings. Expression of miR-193 and miR-210 was investigated by miRNA transient silencing assays using the MCF7, BT20 and MDA-MB-231 cell lines. Functional studies included, xCELLigence system, ApoTox-Glo triplex assay, flow cytometry and transwell inserts were performed to determine cell proliferation, cytotoxicity, apoptosis, migration and invasion, respectively.Results: The most evident effects were associated with cell proliferation after miR-210 silencing in triple negative subtype cell line MDA-MB-231. Using in silico prediction algorithms, TNFRSF10 was identified as one of the potential regulated downstream targets for both miRNAs. The TNFRSF10C and TNFRSF10D mRNA expression inversely correlated with the expression levels of miR-193 and miR210 in breast cell lines and breast cancer patients, respectively. Other potential regulated genes whose expression also inversely correlated with both miRNAs were CCND1, a known mediator on invasion and metastasis, and the tumor suppressor gene RUNX3.Conclusions: In summary, our findings identify miR-193 and miR-210 as potential regulatory miRNA in different molecular subtypes of breast cancer and suggest that miR-210 may have a specific role in MDA-MB-231 proliferation. Our results highlight important new downstream regulated targets that may serve as promising therapeutic pathways for aggressive breast cancers


2016 ◽  
Vol 2 (1) ◽  
pp. 34
Author(s):  
Tarwadi . ◽  
Churiyah . ◽  
Olivia Bunga Pongtuluran ◽  
Fifit Juniarti ◽  
Fery Azis Wijaya

Sambiloto (Andrographis paniculata) banyak digunakan untuk mengobati berbagai penyakit di Indonesia dan negara-negara Asia lainnya. Dalam studi ini, ekstrak metanol dan etanol sambiloto yang diperoleh dari B2PTO Tawangmangu telah diuji terhadap sel lini kanker payudara T47D dan MCF-7 dan sel lini normal fibroblast HFL-1 menggunakan reaksi enzimatik 3-(4,5-dimethylthiazoyl-2-yl) 2,5-diphenyltetrazoliumbromide (MTT). Uji in vitro terhadap sel lini normal fibroblast HFL-1 menunjukkan bahwa 50 ppm ekstrak metanol sambiloto tidak menghambat pertumbuhan sel. Tetapi, ekstrak metanol dan etanolnya menghasilkan IC50 yang relatif rendah pada sel lini kanker payudara, yaitu 111 ppm dan 122 ppm pada sel lini MCF-7 dan 70 ppm dan 197 ppm pada sel lini T47D. Selain itu, campuran ekstrak sambiloto yang mengandung 25% ekstrak Thyponium divaricatum dan Anredera cordifolia memberikan daya hambat pertumbuhan pada sel kanker payudara MCF-7 yang lebih besar, dengan nilai IC50 masing-masing adalah 68 ppm dan 34 ppm. Kesimpulannya, total ekstrak metanol atau etanol sambiloto yang diperoleh dari Tawangmangu memiliki potensi sebagai sumber senyawa anti-kanker serta perlu kajian lebih lanjut.Kata kunci: Ekstrak Andrographis paniculata, MTT, sel lini normal, sel lini kanker, aktivitas anti kanker ABSTRACTSambiloto (Andrographis paniculata) is widely used as medicine to treat various diseases in Indonesia and other Asian countries. In this study, methanolic and ethanolic extracts of sambiloto collected from B2PTO Tawangmangu have been tested againts breast cancer cell lines of T47D and MCF-7 and normal fibroblast cell line of HFL-1 using enzymatic reaction of 3-(4,5-dimethylthiazoyl-2-yl) 2,5-diphenyltetrazoliumbromide (MTT). In vitro assay performed on normal fibroblast of HFL-1 cell line showed that 50 ppm of methanolic extract of sambiloto did not inhibit cell growth. However, methanolic and ethanolic extracts of sambiloto gave relatively low of IC50 on breast cancer cell lines which were 111 ppm and 122 ppm on the MCF-7 cell lines and 70 ppm and 197 ppm on the T47D cell lines, respectively. In addition, the mixture of sambiloto extract containing 25% of Thyponium divaricatum and Anredera cordifolia extracts confered greater growth inhibition on breast cancer cell line of MCF-7, where IC50 values were 68 ppm and 34 ppm, respectively. In conclusion, the total methanolic or ethanolic extract of sambiloto collected from Tawangmangu has potency as a source of anti-cancer compounds and needs further study.Key words: Andrographis paniculata extract, MTT, normal cell line, cancer cell lines, anti-cancer activity


BMC Cancer ◽  
2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Adriane F. Evangelista ◽  
Renato J. Oliveira ◽  
Viviane A. O. Silva ◽  
Rene A. D. C. Vieira ◽  
Rui M. Reis ◽  
...  

Abstract Background Breast cancer is the most frequently diagnosed malignancy among women. However, the role of microRNA (miRNA) expression in breast cancer progression is not fully understood. In this study we examined predictive interactions between differentially expressed miRNAs and mRNAs in breast cancer cell lines representative of the common molecular subtypes. Integrative bioinformatics analysis identified miR-193 and miR-210 as potential regulatory biomarkers of mRNA in breast cancer. Several recent studies have investigated these miRNAs in a broad range of tumors, but the mechanism of their involvement in cancer progression has not previously been investigated. Methods The miRNA-mRNA interactions in breast cancer cell lines were identified by parallel expression analysis and miRNA target prediction programs. The expression profiles of mRNA and miRNAs from luminal (MCF-7, MCF-7/AZ and T47D), HER2 (BT20 and SK-BR3) and triple negative subtypes (Hs578T e MDA-MB-231) could be clearly separated by unsupervised analysis using HB4A cell line as a control. Breast cancer miRNA data from TCGA patients were grouped according to molecular subtypes and then used to validate these findings. Expression of miR-193 and miR-210 was investigated by miRNA transient silencing assays using the MCF7, BT20 and MDA-MB-231 cell lines. Functional studies included, xCELLigence system, ApoTox-Glo triplex assay, flow cytometry and transwell inserts were performed to determine cell proliferation, cytotoxicity, apoptosis, migration and invasion, respectively. Results The most evident effects were associated with cell proliferation after miR-210 silencing in triple negative subtype cell line MDA-MB-231. Using in silico prediction algorithms, TNFRSF10 was identified as one of the potential regulated downstream targets for both miRNAs. The TNFRSF10C and TNFRSF10D mRNA expression inversely correlated with the expression levels of miR-193 and miR210 in breast cell lines and breast cancer patients, respectively. Other potential regulated genes whose expression also inversely correlated with both miRNAs were CCND1, a known mediator on invasion and metastasis, and the tumor suppressor gene RUNX3. Conclusions In summary, our findings identify miR-193 and miR-210 as potential regulatory miRNA in different molecular subtypes of breast cancer and suggest that miR-210 may have a specific role in MDA-MB-231 proliferation. Our results highlight important new downstream regulated targets that may serve as promising therapeutic pathways for aggressive breast cancers


2021 ◽  
Author(s):  
Maryana Teufelsbauer ◽  
Clemens Lang ◽  
Adelina Plangger ◽  
barbara Rath ◽  
Doris Moser ◽  
...  

Abstract Metformin is used to treat patients with diabetes mellitus and that was found to lower the incidence of cancer. The present study investigated the effects of metformin on human bone-derived mesenchymal stromal cells (BM-MSC) and their breast cancer cell line interactions. BM-MSCs were tested for growth stimulation and migration controlling activity on four breast cancer cell lines employing MTT tests, migration scratch tests and assays of the expression of adipokines in Western Blot arrays. Compared to breast cancer cell lines, metformin significantly inhibited the proliferation of BM-MSC lines. Pretreatment of BM-MSCs with metformin showed variable effects on breast cancer cell lines depending on the specific BM-MSC cancer line combination. Metformin significantly impaired the migration of MDA-MB-231 and MDA-MB-436 in response to conditioned media (CM) of drug pretreated BM-MSCs. Metformin-induced alterations of adipokines by BM-MSC CM indicated increased osteogenic signaling and possibly impairment of metastasis. The anticancer activities of metformin seem to be the result of direct and indirect mechanisms. A lower metformin-induced protumor activity of BM-MSCs in the bone microenvironment seem to contribute to the anticancer effects of this drug in breast cancer patients.


2021 ◽  
Author(s):  
Maryam Akbarzadeh

Abstract Background Breast cancer is currently one of the most common types of cancer in women, and metastasis is the first cause of death in breast cancer patients. The epidermal growth factor (EGF) increases the invasion, growth, and migration of cancer cells. In the present study, melatonin, as a natural hormone, in EGF-induced tumor metastasis, was investigated. Methods First, MDA-MB-231 and MCF7 cells were cultured, and then the effects of melatonin on cell viability were determined by MTT assay. Transwell invasion assay was employed to identify the invasiveness of these breast cancer cell lines. Real-time RT-PCR then investigated the expression of MMP9 and MMP2. Cell proliferation was also determined under EGF and melatonin treatment using Ki67 assessment by flow cytometry. Results The rate of invasion and migration of EGF-treated cells increased in both groups, in which melatonin caused increased invasion by EGF in MCF7 cells. MMP9 and MMP2 expression increased significantly in both cell lines under EGF treatment, and melatonin increased these genes' expression in both cell lines (p <0.05). EGF increased the MMP9 and MMP2 gene expression, and melatonin increases EGF-induced expression(p <0.05). The EGF reduced the expression of the Ki67 protein in the MCF7 cell line, which was negatively affected by Melatonin and EGF. In contrast, along with Melatonin, EGF did not affect the proliferation of the MDA-MB-231 cell line. Conclusions Our results show that melatonin, as a natural compound, can increase the effects of EGF in the proliferation, migration, and invasion of cancer cells at low dosages.


Author(s):  
Chul Min Yang ◽  
Tomohiro Chiba ◽  
Bernd Groner

AbstractDistinct gene expression patterns, accompanied by particular epigenetic states, provide the basis for different stages of cellular differentiation. The programming of cells usually proceeds from stem cells to progenitor cells to differentiated progeny. The process, however, is not irreversible, and pluripotency can be reestablished in terminally differentiated cells through the expression of the reprogramming factors (RFs) octamer-binding transcription factor 3/4 (Oct4), sex-determining region Y box 2 (Sox2), Krüppel-like factor 4 (Klf4), and c-Myc. Tumor cell formation is characterized by partial differentiation, epigenetic alterations, and drastic changes in the transcriptional program. As the RF can cause pluripotency through cellular dedifferentiation and epigenetic alterations, it is possible that the activation of their genes might contribute to cellular transformation. The shared capacity for self-renewal between pluripotent stem cells and cancer stem cells is in line with this assumption. The deregulation of RF has been observed in poorly differentiated, high-grade breast cancer and is associated with unfavorable prognosis. Signal transducer and activator of transcription 3 (Stat3) mediates a wide variety of cellular functions including survival, proliferation, and differentiation and is constitutively activated in tumor cells of diverse origins. Stat3 is also accelerates somatic cell reprogramming. We investigated the connection between Stat3 activation and the expression of RF in the breast cancer cell lines MCF-7, SK-BR-3, MDA-MB-231, and MDA-MB-468 and the normal mammary epithelial cell line MCF-10A by real-time quantitative PCR and immunoblot analyses. We detected strong expression of Sox2 and Klf4 messenger RNA (mRNA) in MCF-7 cells and the expression of Oct4 mRNA in all the four cell lines. Immunoblot analyses revealed the strong protein expression of homeobox protein Nanog (Nanog), Oct4, and Sox2 in MCF-7 cells. This cell line only contains a low level of phosphorylated Stat3. We also examined the effect of the Stat3 inhibitor Stattic on the expression of RF and observed that it suppressed mRNA and protein expression of RF in MCF-7 cells. The expression levels of reprogramming proteins can strongly differ from their mRNA expression levels and do not necessarily correspond with the extent of Stat3 activation in the cell lines compared.


2009 ◽  
Vol 2 (3) ◽  
pp. 160-165 ◽  
Author(s):  
Menka Khoobchandani ◽  
B. K. Ojeswi ◽  
Bhavna Sharma ◽  
Man Mohan Srivastava

The present study is aimed to investigate the effects ofChenopodium album(leaves) on the growth of estrogen dependent (MCF-7) and estrogen independent (MDA-MB-468) human breast cancer cell lines. The different solvent extracts (petroleum ether, ethyl acetate and methanol) were assessed for their cytotoxicity using TBE (Trypan blue exclusion) and MTT [3-(4, 5-dimethyl thiazol-2-yl)-2, 5-diphenyl tetrazolium] bioassay. These cells were cultured in MEM (minimum essential medium) medium and incubated with the dilution series of extracts (10–100 mg/ml) in CO2incubator at 37°C for 24 h. Among the various extracts studied for two cell lines, methanolic extract ofC. album(leaves) exhibited maximum antibreast cancer activity having IC50(the concentration of an individual compound leading to 50% inhibition) value 27.31 mg/ml against MCF-7 cell line. Significant percent inhibition (94.06%) in the MeOH extract ofC. album(leaves) at 48 h of exposure and concentration 100 mg/ml (p < 0.05) against MCF-7 breast cancer cell line, indicates the presence of some structural moiety responsible for this observed antiproliferative effect. In vivo study and structural elucidation of its bioactive principle are in progress. Our findings highlight the potential of this plant for its possible clinical use to counteract malignancy development as antibreast cancer bioagent.


2017 ◽  
Vol 2017 ◽  
pp. 1-11 ◽  
Author(s):  
Martha Daniela Mociño-Rodríguez ◽  
Jonnathan Guadalupe Santillán-Benítez ◽  
David Salomón Dozal-Domínguez ◽  
María Dolores Hernández-Navarro ◽  
Miriam Veronica Flores-Merino ◽  
...  

The development of breast cancer is influenced by the adipose tissue through the proteins leptin and adiponectin. However, there is little research concerning AdipoR1 and AdipoR2 receptors and the influence of leptin over them. The objective of this work was to analyze the expression of AdipoR1 and AdipoR2, modulated by differential concentrations of leptin in an obesity model (10 ng/mL, 100 ng/mL, and 1000 ng/mL) associated with breast cancer in MCF-7 and HCC1937 cell lines. Each cell line was characterized through immunohistochemistry, and the expression of AdipoR1 and AdipoR2 was analyzed by PCR in real time using TaqMan® probes. Leptin induced an increase in cell population of MCF-7 (23.8%, 10 ng/mL, 48 h) and HCC1937 (17.24%, 1000 ng/mL, 72 h). In MCF-7, the expression of AdipoR1 decreased (3.81%, 1000 ng/mL) and the expression of AdipoR2 increased by 13.74 times (10 ng/mL) with regard to the control. In HCC1937, the expression of AdipoR1 decreased by 86.28% (10 ng/mL), as well as the expression of AdipoR2 (50.3%, 100 ng/mL). In regard to the results obtained, it could be concluded that leptin has an effect over the expression of AdipoR1 and AdipoR2 mRNA.


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