Tag Archives: breast cancer
Calix[4]arene chalcone amide C-1011 elicits differential effects on the viability of 4T1 mouse breast adenocarcinoma cells with different levels of adaptor protein Ruk/CIN85 expression
L. G. Babich1*, S. G. Shlykov1, O. A. Yesypenko2, A. O. Bavelska-Somak1,
A. G. Zahoruiko1, I. R. Horak1, L. B. Drobot1, S. O. Kosterin1
1Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
2Institute of Organic Chemistry, National Academy of Sciences of Ukraine, Kyiv;
*e-mail: babich@biochem.kiev.ua
Received: 07 February 2021; Accepted: 01 July 2022
According to our earlier data, calix[4]arene chalcone amides modulate Ca ions exchange in the myometrium mitochondria and the level of inner membrane polarization that can potentially affect cell survival. To test this hypothesis, we studied the effect of calix[4]arene with 4 chalcone amide groups on mitochondria membrane polarization and viability of 4T1 mouse breast adenocarcinoma cells, a surrogate model of human triple-negative breast cancer, and on its highly malignant subline overexpressing the adaptor protein Ruk/CIN85. Mitochondria membrane potential was measured by flow cytometry, and cell viability was assessed using Trypan blue dye exclusion. It was shown that mitochondrial membranes of control (Mock) cells had a higher polarization level (67.80 ± 8.82 r.u., n = 5) compared to 4T1 cells with up-regulation of Ruk/CIN85 (RukUp cells) (25.42 ± 2.58 r.u., n = 4). Upon incubation of cells with 1 μM calix[4]arene C-1011, the CCCP-sensitive component of mitochondrial membranes polarization decreased (by almost 50%) in 4T1 Mock cells and did not change in RukUp cells compared with the control. It was demonstrated that 1 μM calix[4]arene C-1011 suppressed the viability of 4T1 Mock cells by 45%, but did not affect RukUp cells considerably. It was suggested that calix[4]arene chalcone amide С-1011 decreased mouse breast adenocarcinoma 4T1 cell viability at least by affecting mitochondrial membrane polarization.The data obtained indicate the prospects of further studies of calix[4]arene chalcone amide as a potential anticancer drug candidate.
Extracellular vesicles produced by mouse breast adenocarcinoma 4T1 cells with up- or down-regulation of adaptor protein Ruk/CIN85 differentially modulate the biological properties of 4T1 WT cells
A. Yu. Zhyvolozhnyi1,2*, I. R. Horak1, D. S. Geraschenko1, M. O. Gomozkova3,
O. O. Hudkova1, S. J. Vainio2, A. A. Samoylenko2, L. B. Drobot1
1Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
2Faculty of Biochemistry and Molecular Medicine, University of Oulu, Finland;
3Brigham Young University-Idaho, Rexburg, USA;
*e-mail: ppndl2@gmail.com
Received: 02 November 2021; Accepted: 12 November 2021
Extracellular vesicles (EVs) are secreted by most cell types under both physiological and pathological conditions and were proposed to be actively involved in intercellular communication. The mode of EVs action is dependent on their cargos composition. EVs play an important role in tumor initiation, recurrence, metastasis and therapeutic resistance. EVs marker proteins Alix and Tsg101 and cortactin are the binding partners of adaptor protein Ruk/CIN85. The present study aims to analyze the regulatory effects of EVs produced by 4T1 cells with overexpression (RukUp) or down-regulation (RukDown) of adaptor protein Ruk/CIN85 on proliferation rate, migration and invasion activity of parental 4T1 WT cells. EVs from conditioned medium of 4T1 RukUp or RukDown cells were isolated by differential centrifugation followed by further purification using Exo-spin™ kit (Cell Guidance Systems). The number and size of EVs were characterized by NTA (Malvern Panalytical NanoSight NM300) instrument. The content of marker proteins and Ruk/CIN85 in isolated EVs was analyzed by Western-blotting. The viability, migration and invasion activity of 4T1 WT cells were studied using MTT-test, scratch-test and Boyden chamber assay, respectively. It was demonstrated for the first time that adaptor protein Ruk/CIN85 is a constitutive component of EVs produced by 4T1 cells. It was also shown that EVs produced by 4T1 cells with different levels of Ruk/CIN85 expression are characterized by a specific profile of the content of its multiple molecular forms. It turned out that the ability of EVs to modulate the proliferative activity, motility and invasiveness of 4T1 WT cells was tightly correlated with the biological properties of 4T1 cells that produce EVs (highly aggressive 4T1 RukUp cells or weakly invasive 4T1 RukDown cells). Our data suggest that adaptor protein Ruk/CIN85 is not only a constitutive component of cargos composition of EVs produced by tumor cells but, depending on its content in EVs, plays an active role in the control of carcinogenesis.
Effect of IRAK1/4 inhibitor on IL-1β, IL-6, INF-γ and TNF-α expression in breast cancer cells of several lines
M. Rezaei1, B. Shahouzehi2,4, S. Rahemi1,3, H. Fallah1*, M. Salarkarimi1
1Department of Clinical Biochemistry, Afzalipour School of Medicine, Kerman University of Medical Sciences, Kerman, Iran;
2Cardiovascular Research Center, Institute of Basic and Clinical Physiology Sciences, Kerman University of Medical Sciences, Kerman, Iran;
3Physiology Research Center, Institute of Basic and Clinical Physiology Sciences, Kerman University of Medical Sciences, Kerman, Iran;
4Student Research Committee, School of Medicine, Kerman University of Medical Sciences, Kerman, Iran;
*e-mail: hf59ma@gmail.com
Received: 28 July 2020; Accepted: 07 July 2021
Recent studies have shown that inflammation mediated via interleukin-1 receptor-associated kinases (IRAKs) is associated with cancer cells drug resistance. We aimed to evaluate the expression of inflammatory cytokines as the potential mechanism involved in the development of cancer cells resistance to conventional chemotherapy drugs. Breast cancer cells of BT549, BT20 and MB468 lines were treated with IRAK 1/4 inhibitor alone or in combination with chemotherapeutic agents methotrexate and topotecan. Expression of IL-1β, IL-6, TNF-α, and IFN-γ genes was quantified by real-time PCR. It was found that IRAK1/4 inhibitor suppressed IL-1β expression in BT549 cells at most and had minimal effect on IL-6 expression in MB468 cells. For the first time we showed that concomitant use of IRAK1/4 inhibitor with topotecan and methotrexate reduced IL-1β, IFN γ, TNF-α and IL-6 expression in BT-20, BT-549, MB-468 cell lines compared to the controls. It is suggested that specific IRAK inhibitors in combination with conventional chemotherapy can be used in cancer treatment to increase drug sensitivity and decrease the risk of tumor recurrence.
miR-329-containing exosomes derived from breast tumor cells suppress VEGF and KDM1A expression in endothelial cells
N. Maleki1,2,3*, F. Karami1, S. Heyati2, M. HadiZadeh3, Gh. Parnian4*
1Department of Cellular and Molecular Biology, Faculty of Biological Sciences, Islamic Azad University-Tehran North Branch, Tehran, Iran;
*e-mail: dr.nargesmaleki@yahoo.com;
2Gynecology and reproductive biology Department, Kowsar poly-clinic, Tehran, Iran;
3Cancer Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran;
4Appletree Medical group, 275 Dundad W (Grange), Toronto, Ontario, Canada;
*e-mail: ghazalehparnian1@gmail.com
Received: 03 February 2021; Accepted: 07 July 2021
The exosomal transfer of miRNAs from tumor cells is considered to modulate VEGF expression and angiogenesis in endothelial cells. The aim of our investigation was to focus exclusively on the ability of specific exosomal miR329 to regulate angiogenesis within breast tumor. All experiments were done on MCF-7 and HUVEC cell lines. Exosomes were derived from MCF-7 cells both untreated and treated with tamoxifen that is an effecrive suppressor of hormone receptor-positive breast cancer. The level of miR32 and its targeted genes VEGF and lysine (K)-specific demethylase 1A (KDM1A) expression was estimated with q-RT-PCR. The PKH26 red fluorescent labeling kit was used to label the isolated exosomes and monitor their uptake. It was shown that the relative amount of miR-329 in exosomes was twice as large as in breast cancer cells. Fluorescence microscopy imaging presented that exosomes from MCF-7 cells were able to penetrate into endothelial cells and concentrate in the cytoplasm. It was observed that exosomes derived from untreated breast cancer cells induced KDM1A and VEGF gene expressions whereas exosomes from tamoxifen-treated cancer cells induced time-dependent decrease of KDM1A and VEGF expression in endothelial cells. It is assumed that the transfer of miR-329 containing exosomes from tamoxifen treated breast cancer cells to the endothelial cells could repress angiogenic molecular signaling pathway and be used as a supplementary strategy in breast cancer treatment.
Adaptor protein Ruk/CIN85 affects redox balance in breast cancer cells
I. R. Horak*, N. V. Latyshko, O. O. Hudkova, T. O. Kishko,
O. V. Khudiakova, D. S. Gerashchenko, T. D. Skaterna,
I. P. Krysiuk, S. G. Shandrenko, L. B. Drobot
Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
*e-mail: iryna.horak@gmail.com
Received: 25 February 2020; Accepted: 15 May 2020
Excessive reactive oxygen species (ROS) production may lead to damage of cellular proteins, lipids and DNA, and cause cell death. Our previous findings demonstrated that increased level of adaptor protein Ruk/CIN85 contributes to breast cancer cells malignancy. The aim of this study was to investigate the role of Ruk/CIN85 in the maintaining of the redox balance in cancer cells. Mouse breast adenocarcinoma 4T1 cells with different levels of Ruk/CIN85 expression were used as a model in this study. Activities of catalase (CAT), glutathione peroxidase (GPx), superoxide dismutase (SOD), aldehyde dehydrogenase (ALDH) and formaldehyde dehydrogenase (FALDH), as well as H2O2 and aldehydes content were measured using fluorometric assays. Gene expression correlations between Ruk/CIN85 and antioxidant enzymes in breast cancer samples were analyzed using ist.medisapiens transcriptomic database. It was demonstrated that Ruk/CIN85-overexpressing 4T1 cells were characterized by increased production of H2O2 and reduced activities of CAT, GPx and SOD. Overexpression of Ruk/CIN85 resulted in decreased content of aldehydes together with increased activity of ALDH, while in Ruk/CIN85-knocked down 4T1 cells, activities of ALDH and FALDH were decreased. The data of transcriptomic analysis revealed the correlations between SH3KBP1 expression and CAT, GPX4, ALDH1A1, ALDH1L1, ALDH2, GSR, SOD1 in human breast carcinomas samples. The obtained results indicate that adaptor protein Ruk/CIN85 affects redox balance in mouse breast adenocarcinoma 4T1 cells.
Adaptor protein Ruk/CIN85 modulates resistance to doxorubicin of murine 4T1 breast cancer cells
I. R. Horak, D. S. Gerashchenko, L. B. Drobot
Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
e-mail: iryna.horak@gmail.com
The acquisition of chemoresistance in the course of tumor progression includes activation of membrane ABC transporters, detoxification enzymes, cell cycle deceleration and activation of specific signaling pathways such as Akt/mTOR, MAPK, NF-κB. Adaptor proteins play an essential role in the assembly of supramolecular signaling complexes, maintaining and directing the intracellular signaling. One of such proteins, called Ruk/CIN85, is strongly associated with malignant transformation and metastasis. In present study we investigated the Ruk/CIN85 effect of up/down-regulation on the transforming potential and doxorubicin resistance of highly aggressive mouse breast adenocarcinoma 4T1 cells. It was demonstrated that 4T1 cells overexpressing Ruk/CIN85 possessed increased resistance to doxorubicin (in the range of concentrations 0.1–10.0 µM) while knockdown cells were the most sensitive. Also, high levels of Ruk/CIN85 in 4T1 cells positively correlated with their ability to form colonies in semi-solid agar. Ruk/CIN85-overexpressing cells formed four times more colonies in comparison with Ruk/CIN85 nockdown cells, the growth of which revealed higher resistance to doxorubicin action.
Biological effects of thyroid hormones
T. S. Saatov, A. A. Abduavaliev
Institute of Bioorganic Chemistry, Uzbekistan Academy of Sciences, Tashkent;
e-mail: t.saatov@yandex.ru
The article presents the findings from the study on multifunctional effects of thyroid hormones in relation to normal and malignantly transformed tissues and cells. Both “rapid” and «slow» effects of thyroid hormones including calorigenic effects and effects over adenylate cyclase – cAMP system have been described. Thyroxin (Т4) has been established capable to inhibit proliferation and to induce apoptosis of cells carrying Т4 receptors on their membranes as well as to change course of metabolic processes under its effect. Spectrum of Т4 targets is quite broad to include not only cells of hormone-producing organs, to name those of the breast and the colon, but also other types of cells to name melanin-containing ones; Т4 effects resulting in reconstruction of presentation of regulatory proteins on the cell membrane surface to ultimately activate the process of cell apoptosis. Our findings help determine alternative paths for hormonal regulation of cell proliferation and apoptosis of cells of hormone-dependent tumors, breast cancer, in particular, upon impossibility to regulate the processes by conventional methods. This facilitates understanding mechanisms for activation of signal system of the breast cancer’s cells by hormones upon changes in expression of receptors on the cells’ surface, making possible development of novel strategy for replacement therapy of hormone-dependent tumors upon low efficacy of drug therapy.