-
Dipeptidyl-Aminopeptidases 8 and 9 Regulate Autophagy and Tamoxifen Response in Breast Cancer Cells.Cells Aug 2023The cytosolic dipeptidyl-aminopeptidases 8 (DPP8) and 9 (DPP9) belong to the DPPIV serine proteases with the unique characteristic of cleaving off a dipeptide...
The cytosolic dipeptidyl-aminopeptidases 8 (DPP8) and 9 (DPP9) belong to the DPPIV serine proteases with the unique characteristic of cleaving off a dipeptide post-proline from the -termini of substrates. To study the role of DPP8 and DPP9 in breast cancer, MCF-7 cells (luminal A-type breast cancer) and MDA.MB-231 cells (basal-like breast cancer) were used. The inhibition of DPP8/9 by 1G244 increased the number of lysosomes in both cell lines. This phenotype was more pronounced in MCF-7 cells, in which we observed a separation of autophagosomes and lysosomes in the cytosol upon DPP8/9 inhibition. Likewise, the shRNA-mediated knockdown of either DPP8 or DPP9 induced autophagy and increased lysosomes. DPP8/9 inhibition as well as the knockdown of the DPPs reduced the cell survival and proliferation of MCF-7 cells. Additional treatment of MCF-7 cells with tamoxifen, a selective estrogen receptor modulator (SERM) used to treat patients with luminal breast tumors, further decreased survival and proliferation, as well as increased cell death. In summary, both DPP8 and DPP9 activities confine macroautophagy in breast cancer cells. Thus, their inhibition or knockdown reduces cell viability and sensitizes luminal breast cancer cells to tamoxifen treatment.
Topics: Humans; Tamoxifen; Autophagy; Macroautophagy; MCF-7 Cells; Aminopeptidases; Neoplasms
PubMed: 37626841
DOI: 10.3390/cells12162031 -
International Journal of Oncology Mar 2021Ampelopsis megalophylla has been found to demonstrate anticancer activities in human cancer cells; however, the effect of total flavone extract (TFE), commonly used in...
Ampelopsis megalophylla has been found to demonstrate anticancer activities in human cancer cells; however, the effect of total flavone extract (TFE), commonly used in Traditional Chinese Medicine, remains unclear. Furthermore, there is limited information on its effects on breast cancer cell lines. The present study aimed to investigate the inhibitory effects of TFE in different human cancer cell lines. In addition, the underlying mechanisms and the signaling pathways involved were also investigated by determining tumor cell morphological changes, and differences in the cell cycle, apoptosis, mitochondrial transmembrane potential, and related protein expression levels in a breast cancer cell line. It was found that TFE inhibited proliferation in seven cancer cell lines (HeLa, A549, MCF‑7, HepG2, A2780, SW620 and MDA‑MB‑231 and demonstrated a strong inhibitory effect on MCF‑7 cell proliferation. Cell morphological changes were also observed and arrested at the G2/M phase following treatment with TFE at different concentrations. In addition, TFE disrupted the mitochondrial membrane potential and upregulated the expression level of apoptotic proteins, including caspase‑3, ‑8 and ‑9, the Bax/Bcl‑2 ratio, and Apaf‑1 in time‑dependent manner. These results indicated that TFE induced apoptosis of the MCF‑7 cells via a mitochondrial‑mediated apoptotic pathway. In conclusion, TFE is potentially effective in treating breast cancer.
Topics: Ampelopsis; Antineoplastic Agents, Phytogenic; Apoptosis; Breast Neoplasms; Female; Humans; MCF-7 Cells; Mitochondria; Plant Extracts
PubMed: 33469684
DOI: 10.3892/ijo.2021.5172 -
Biological & Pharmaceutical Bulletin 2022Solid tumors habitually harbor regions with insufficient oxygen away from vasculature. Hypoxia is an important factor that confers malignant phenotypes like...
Solid tumors habitually harbor regions with insufficient oxygen away from vasculature. Hypoxia is an important factor that confers malignant phenotypes like chemoresistance to tumor cells. We have demonstrated that cathepsin G (CG) stimulates cell aggregation in breast cancer MCF-7 cells by activating insulin-like growth factor-1 signaling. We investigated whether cancer cell aggregates induced by CG acquire hypoxia-dependent chemoresistance. Pimonidazole staining and hypoxia-inducible factor (HIF)-1α and -2α expression indicated that the core of the cell aggregates was hypoxic. Electrophoretic mobility shift and reporter assays showed that the CG-induced cell aggregates displayed transcriptional activity through HIF-responsive elements. Moreover, HIF target genes PGK1 and SLC2A1 demonstrated upregulated expression in CG-induced cell aggregates, indicating that the aggregates expressed functional HIF. Doxorubicin (DXR)-induced cytotoxicity was significantly lower in the cell aggregates induced by CG compared with monolayer cells under normoxia. Unexpectedly, the upregulation of P-glycoprotein expression, which is reported to be a HIF-target gene, and decreasing intracellular accumulation of DXR was not detected in the cell aggregates as opposed to in monolayer cells under normoxia. Additionally, reduction of DXR sensitivity in the aggregates was not suppressed by treatment with the HIF inhibitor, YC-1 and HIF-1α small interfering RNA (siRNA). Therefore, we conclude that cell aggregation induced by CG decreases DXR sensitivity via a HIF-independent mechanism.
Topics: Humans; Cathepsin G; MCF-7 Cells; Doxorubicin; Cell Aggregation; RNA, Small Interfering; Hypoxia; Neoplasms
PubMed: 36450530
DOI: 10.1248/bpb.b22-00447 -
International Journal of Environmental... Jun 2021Contaminants of Emerging Concern (CECs) with estrogenic or estrogenic-like activity have been increasingly detected in aquatic environments and have been an issue of...
Contaminants of Emerging Concern (CECs) with estrogenic or estrogenic-like activity have been increasingly detected in aquatic environments and have been an issue of global concern due to their potential negative effects on wildlife and human health. This study used the MCF-7 cell proliferation assay (E-Screen) to assess the estrogenic activity profiles in Maryland Coastal Bays (MCBs), a eutrophic system of estuaries impacted by human activities. Estrogenic activity was observed in all study sites tested. Water samples from MCBs increased MCF-7 cell proliferation above the negative control from 2.1-fold at site 8, located in Sinepuxent Bay close to the Ocean City Inlet, to 6.3-fold at site 6, located in Newport Bay. The proliferative effects of the sediment samples over the negative control ranged from 1.9-fold at the Assateague Island National Seashore site to 7.7-fold at the Public Landing site. Moreover, elevated cell proliferation ( < 0.05) was observed when cells were co-exposed with 17ß-Estradiol (E2), while reduction in cell proliferation was observed when cells were co-exposed with the antagonist ICI 182, 780 suggesting that cell proliferative effects were primarily mediated by the estrogen receptor (ER). These results suggest the occurrence of some estrogenic or hormonal-like compounds in the MCBs and are consistent with our previous findings based on vitellogenin analyses.
Topics: Bays; Cell Proliferation; Estrogens; Humans; MCF-7 Cells; Maryland
PubMed: 34207818
DOI: 10.3390/ijerph18126254 -
Experimental Oncology Aug 2022Oncolytic virotherapy is an emerging biotherapeutic platform for selectively infecting cancer cells and triggering apoptosis in a number of malignant cells due to robust...
UNLABELLED
Oncolytic virotherapy is an emerging biotherapeutic platform for selectively infecting cancer cells and triggering apoptosis in a number of malignant cells due to robust viral replication. Studies related to the oncolytic activity of human orthopneumovirus (hOPV) are conflicting.
AIM
This study was designed to elucidate the possible role of hOPV in the modulation of cell growth and apoptosis in cancer cell lines including human epidermoid carcinoma (HEp-2), lung epithelial cell line (A549), and breast cancer cell line (MCF-7).
MATERIALS AND METHODS
The oncolytic activity of hOPV on cancer cells was studied in vitro. The virus titers were determined by tissue culture infectious dose (TCID50/mL) in A549 cell. The cytotoxic effect of the virus on HEp-2, A549, and MCF-7 was determined using MTT and trypan blue dye exclusion test assays. hOPV in the infected cells was detected using real-time reverse transcription polymerase chain reaction (rRT-PCR) and indirect immunofluorescence (IIF) assays. The relative expression of apoptosis-related genes (CASP-3, -8, -9, Bax, Bcl-2, Bcl-XL, TP53, P21) during virus infection was estimated using rRT-PCR assay in comparison with the house-keeping gene (GAPDH).
RESULTS
hOPV infection inhibited the growth of HEp-2, A549, and MCF-7 cells in a dose-and time-dependent manner. At a multiplicity of infection (MOI) of 5, hOPV reduced the viability of A549 cells to about 16%, HEp-2 to 22%, and MCF-7 to 28% (p = 0.001), while no significant inhibitory effect was observed when cells were infected at MOI of 1 and 2. hOPV mRNA and antigens were detected in infected HEp-2, A549, and MCF-7 cells by RT-PCR and IIF. Upon hOPV infection, expression of CASP-3, -8, -9, as well as Bax, TP53, and p21 mRNA increased while expression of Bcl-2, Bcl-xL anti-apoptotic genes decreased. In hOPV-infected A549 cells, the fold increase of CASP-8 and CASP-9, Bax, TP53, and P21 expression exceeded significantly compared to that in HEp-2 or MCF-7 cells.
CONCLUSIONS
Our results provide evidence that hOPV could be a potential candidate for oncolytic virotherapy.
Topics: Apoptosis; Humans; MCF-7 Cells; Neoplasms; Proto-Oncogene Proteins c-bcl-2; RNA, Messenger; bcl-2-Associated X Protein
PubMed: 35964639
DOI: 10.32471/exp-oncology.2312-8852.vol-44-no-2.18084 -
Journal of the Egyptian National Cancer... Dec 2022Mammosphere formation assay has become a versatile tool to quantify the activity of putative breast cancer stem cells in non-adherent in vitro cultures. However,...
BACKGROUND
Mammosphere formation assay has become a versatile tool to quantify the activity of putative breast cancer stem cells in non-adherent in vitro cultures. However, optimizing the suspension culture system is crucial to establish mammosphere cultures from primary breast tumors.
METHODS
This study aimed at determining the self-renewal and sphere-forming potential of breast cancer stem-like cells derived from human primary invasive ductal carcinoma and normal breast tissue samples, and MCF-7 breast cancer cell line using an optimal suspension culture system. Mammosphere-forming efficiency of the mammospheres generated from the tissue samples and cell line were compared. We evaluated the expression of CD44/CD24/ and CD49f/EpCAM/ phenotypes in the stem-like cells by flow cytometry. CK-18, CK-19, α-SMA, and EpCAM marker expression was assessed using immunohistochemical staining.
RESULTS
Breast epithelial cells isolated from the three samples formed two-dimensional spheroids in suspension cultures. Interestingly, mammospheres formed from patient-derived primary breast tumors were enriched in breast cancer stem-like cells with the phenotype CD44/CD24/ and exhibited a relatively more number of large spheres when compared to the normal breast stem cells. MCF-7-derived SCs were more aggressive and resulted in the formation of a significantly higher number of spheroids. The expression of CK-18/CK-19 and α-SMA/EpCAM proteins was confirmed in breast cancer tissues.
CONCLUSIONS
Thus, the use of primary tumor specimens and breast cancer cell lines as suitable models for elucidating the breast cancer stem cell activity was validated using mammosphere culture system.
Topics: Humans; Female; MCF-7 Cells; Breast; Breast Neoplasms; Neoplastic Stem Cells
PubMed: 36504339
DOI: 10.1186/s43046-022-00152-1 -
Nutrition and Cancer 2021Carotenoids found in fruits and vegetables are compounds with significant biological activities. Epidemiological studies report that these compounds have significant...
Carotenoids found in fruits and vegetables are compounds with significant biological activities. Epidemiological studies report that these compounds have significant anticancer effects, as well reducing the risk of cancer. In the present study, we aimed to determine the effects of capsanthin, an important carotenoid of paprika, on expressions of proteins playing roles in the mitochondrial apoptosis pathway, in addition to its possible cytotoxic and genotoxic effects in MCF-7 cells. Furthermore, possible oxidant/anti-oxidant roles of capsanthin on MCF-7 cells were investigated. The viability of MCF-7 cells was significantly decreased after 24 h of capsanthin application. After Comet analysis, it was determined that the capsanthin caused DNA damage on a dose-dependent manner. Furthermore, Western blot analysis showed that capsanthin application increased p53 and Bax protein expressions and caused a decrease in Bcl-2 protein level. Capsanthin treatment decreased catalase and glutathione levels but increased lipid peroxidation. These results show that the capsanthin causes oxidative stress and DNA damage, and increases mitochondrial apoptotic mechanism-mediated cell death after p53 and Bax protein activations.
Topics: Apoptosis; DNA Damage; Humans; MCF-7 Cells; Xanthophylls
PubMed: 32933334
DOI: 10.1080/01635581.2020.1819347 -
Journal of Gastrointestinal Cancer Dec 2019The importance of earthworm in treatment of various diseases has been proven in ancient literatures. Nowadays, with advances in biotechnology, earthworm is considered a...
BACKGROUND AND AIM
The importance of earthworm in treatment of various diseases has been proven in ancient literatures. Nowadays, with advances in biotechnology, earthworm is considered a rich natural source of many biomolecules that possesses antioxidant and antitumor activities. The present study aimed to evaluate the antitumor activity of earthworm powder (Lumbricus terrestris) against two cell lines, breast cancer cells (MCF-7) and prostate cancer cells (PC-3).
METHODS
Fully matured earthworms (L. terrestris) were collected from soil in Baghdad, Iraq. To assess the cytotoxicity of earthworm powder, the MTT assay was conducted on cancerous (MCF-7 and PC-3 cells) and normal cell line (WRL68 cells) lines.
RESULTS
It was revealed that earthworm powder exerts cytotoxic effects against two cancer cell lines. The viability of MCF-7 and PC-3 cells decreased with increasing the concentration. The IC50 values for PC-3 and MCF-7 cell lines were 265.5 and 965.9 μg/ml, respectively, while the earthworm powder exhibited no cytotoxicity against the WRL68 cells. According to the analysis of the results of the multiple cytotoxicity assay (HCS), the treatment of PC-3 cells with 100, 200, and 400 μg/ml of earthworm powder for 24 h at 37 °C led to cell death by changing the permeability of mitochondrial membrane resulting in cytochrome C release and inducing apoptosis.
CONCLUSION
The results of the present study contribute additional evidence for the antitumor activity of earthworm extracts. Therefore, further research should concentrate on isolating and identifying the earthworm's active biomolecules that have antitumor activity by investigating the molecular mechanism, genetics, and pathways responsible for the antitumor activity of these biomolecules.
Topics: Animals; Antineoplastic Agents; Cell Proliferation; Cell Survival; Drug Screening Assays, Antitumor; Humans; Iraq; MCF-7 Cells; Neoplasms; Oligochaeta; PC-3 Cells; Powders; Tissue Extracts
PubMed: 31321664
DOI: 10.1007/s12029-019-00268-z -
International Journal of Molecular... Feb 2022To target breast cancer (BC), epigenetic modulation could be a promising therapy strategy due to its role in the genesis, growth, and metastases of BC. Valproic acid...
To target breast cancer (BC), epigenetic modulation could be a promising therapy strategy due to its role in the genesis, growth, and metastases of BC. Valproic acid (VPA) is a well-known histone deacetylase inhibitor (HDACi), which due to its epigenetic focus needs to be studied in depth to understand the effects it might elicit in BC cells. The aim of this work is to contribute to exploring the complete pharmacological mechanism of VPA in killing cancer cells using MCF-7. LC-MS/MS metabolomics studies were applied to MCF-7 treated with VPA. The results show that VPA promote cell death by altering metabolic pathways principally pentose phosphate pathway (PPP) and 2'deoxy-α-D-ribose-1-phosphate degradation related with metabolites that decrease cell proliferation and cell growth, interfere with energy sources and enhance reactive oxygen species (ROS) levels. We even suggest that mechanisms such as ferropoptosis could be involved due to deregulation of L-cysteine. These results suggest that VPA has different pharmacological mechanisms in killing cancer cells including apoptotic and nonapoptotic mechanisms, and due to the broad impact that HDACis have in cells, metabolomic approaches are a great source of information to generate new insights for this type of molecule.
Topics: Apoptosis; Breast Neoplasms; Chromatography, Liquid; Female; Histone Deacetylase Inhibitors; Humans; MCF-7 Cells; Metabolomics; Tandem Mass Spectrometry; Valproic Acid
PubMed: 35269790
DOI: 10.3390/ijms23052645 -
Molecules (Basel, Switzerland) May 2022Novel 4-amino-thieno[2,3-d]pyrimidine-6-carboxylates substituted at the second position were prepared by cyclocondensation of 2-amino-3-cyano-thiophene and aryl nitriles...
Novel 4-amino-thieno[2,3-d]pyrimidine-6-carboxylates substituted at the second position were prepared by cyclocondensation of 2-amino-3-cyano-thiophene and aryl nitriles in an acidic medium. The design of the target compounds was based on structural optimization. The derivatives thus obtained were tested in vitro against human and mouse cell lines. The examination of the compound effects on BLAB 3T3 and MFC-10A cells showed that they are safe, making them suitable for subsequent experiments to establish their antitumor activity. The photoirritancy factor of the compounds was calculated. Using the MTT test, the antiproliferative activity to MCF-10A, MCF-7 and MDA-MB-231 cell lines was estimated. The best antiproliferative effect in respect to the MCF-7 cell line revealed compound 2 with IC 4.3 ± 0.11 µg/mL (0.013 µM). The highest selective index with respect to MCF-7 cells was shown by compound (SI = 19.3), and to MDA-MB-231 cells by compound (SI = 3.7). Based on energy analysis, the most stable conformers were selected and optimized by means of density functional theory (DFT). Ligand efficiency, ligand lipophilicity efficiency and the physicochemical parameters of the target 4-amino-thienopyrimidines were determined. The data obtained indicated that the lead compound among the tested substances is compound .
Topics: Animals; Antineoplastic Agents; Breast Neoplasms; Carboxylic Acids; Cell Line, Tumor; Female; Humans; Ligands; MCF-7 Cells; Mice; Pyrimidines
PubMed: 35630793
DOI: 10.3390/molecules27103314