BACKGROUND: Triple negative breast cancer (TNBC) is a particularly aggressive and difficult-to-treat subtype of breast cancer that requires the development of novel therapeutic strategies. To pave the way for such developments it is essential to characterize new molecular players in TNBC. MicroRNAs (miRNAs) constitute interesting candidates in this regard as they are frequently deregulated in cancer and contribute to numerous aspects of carcinogenesis. METHODS AND RESULTS: Here, we discovered that miR-4649-5p, a miRNA yet uncharacterized in breast cancer, is associated with better overall survival of TNBC patients. Ectopic upregulation of the otherwise very low endogenous expression levels of miR-4646-5p significantly decreased the growth, proliferation, and migration of TNBC cells. By performing whole transcriptome analysis and physical interaction assays, we were able to identify the phosphatidylinositol phosphate kinase PIP5K1C as a direct target of miR-4649-5p. Downregulation or pharmacologic inhibition of PIP5K1C phenocopied the growth-reducing effects of miR-4649-5p. PIP5K1C is known to play an important role in migration and cell adhesion, and we could furthermore confirm its impact on downstream PI3K/AKT signaling. Combinations of miR-4649-5p upregulation and PIP5K1C or AKT inhibition, using the pharmacologic inhibitors UNC3230 and capivasertib, respectively, showed additive growth-reducing effects in TNBC cells. CONCLUSION: In summary, miR-4649-5p exerts broad tumor-suppressive effects in TNBC and shows potential for combined therapeutic approaches targeting the PIP5K1C/PI3K/AKT signaling axis.
- MeSH
- fosfatidylinositol-3-kinasy metabolismus MeSH
- lidé MeSH
- mikro RNA * genetika metabolismus MeSH
- nádorové buněčné linie MeSH
- pohyb buněk genetika MeSH
- proliferace buněk genetika MeSH
- protoonkogenní proteiny c-akt genetika metabolismus MeSH
- regulace genové exprese u nádorů MeSH
- triple-negativní karcinom prsu * farmakoterapie genetika metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- MeSH
- imunoterapie ekonomika MeSH
- lidé MeSH
- onkologická péče - zařízení trendy MeSH
- triple-negativní karcinom prsu * genetika komplikace terapie MeSH
- zachování plodnosti metody MeSH
- Check Tag
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- zprávy MeSH
miRNA expression in triple-negative breast cancers (TNBC) has mainly been studied from a methodological viewpoint. However, it has not been considered that miRNA expression profile may be associated with a specific morphological entity inside every tumor. The verification of this hypothesis on a set of 25 TNBCs was the subject of our previous work, where we confirmed specific expression of the studied miRNAs in 82 samples of different morphologies including inflammatory infiltrate, spindle cell, clear cell, and metastases after RNA extraction and purification as well as microchip and biostatistical analysis. In the current work, we demonstrate a low suitability of in situ hybridization method for miRNA detection compared to RT-qPCR, and in detail discuss the biological role of 8 miRNAs with the most significant changes of expression.
- MeSH
- lidé MeSH
- mikro RNA * genetika metabolismus MeSH
- triple-negativní karcinom prsu * genetika patologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Breast cancer metastases are the main reason for women ́s highest cancer mortality. Even though tumor cell dissemination via circulating tumor cells (CTC) released from the primary site is a very ineffective process, distant metastases appear in 46% of triple-negative breast cancer (TNBC) patients corresponding to the disease aggressiveness. Laboratory models for functional testing which mimic the spread of metastatic cells are needed for efficient investigation of the underlying mechanisms and therapeutic intervention. Here, we describe novel isogenic variants LMC3 and CTC3 of human TNBC cell line MDA-MB-231 that were derived by repeated injection of tumor cells into the tail vein of immunodeficient mice and subsequent selection of metastatic cells from lung metastases. These variants have increased migration potential, altered expression profiles, and elevated tumorigenic potential. Moreover, cell line CTC3 readily produces metastases in the lungs and bone marrow and detectable viable circulating tumor cells in the blood. This model enables rapid and cost-efficient strategies for biomarker exploration and novel intervention approaches to limit the CTC presence in the blood and hence tumor dissemination.
- MeSH
- biologické markery MeSH
- lidé MeSH
- metastázy nádorů MeSH
- myši MeSH
- nádorové buněčné linie MeSH
- nádorové cirkulující buňky * metabolismus MeSH
- nádory plic * sekundární MeSH
- triple-negativní karcinom prsu * genetika patologie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Receptors of the large HER family play an important role in breast cancer, which is undergoing a gradual development in connection with biological development, both in the fi eld of diagnostics and therapy. Dimerization of HER-2 with other HER members, such as HER-3, is the biggest driver of tumor cell growth and survival. Numerous studies show that HER-3 gene overexpression correlates with poor prognosis. However, other studies have shown HER-3 overexpression to be a positive prognostic factor. HER-3 may confer resistance to certain EGFR or HER-2 receptor therapeutics. An interesting fact, however, is that HER-3 expression can serve as a marker in immunotherapy for triple-negative breast cancer (TNBC). It is thought to be involved not only in cell survival and proliferation, but also in the regulation of PD-L1 expression. In breast cancer, PD-L1 expression is heterogeneous and is generally associated with the presence of tumor-infi ltrating lymphocytes and a number of factors with poor prognosis such as young age, hormone receptor negativity, and high HER-2 expression and proliferation index. Our results showed amplifi cation of HER-3 (CERB3) in 2 out of a sample of 20 patients with TNBC, and 13 of 20 HER-2-positive patients. PD-L1 expression was demonstrated in 3 out of 13 HER-3-positive patients and 2 out of 2 HER-3-positive TNBC patients. There was a strong correlation between positive HER-3 and PD-L1 TNBC expression (p = < 0.001). Thus, the view of the HER-3 receptor will be much more complex, and the overexpression of this receptor appears to have both negative and positive prognostic and clinical impacts (Tab. 1, Ref. 17).
- MeSH
- amplifikace genu MeSH
- antigeny CD274 biosyntéza MeSH
- cytogenetické vyšetření MeSH
- dospělí MeSH
- erbB receptory škodlivé účinky MeSH
- imunoanalýza MeSH
- lidé středního věku MeSH
- lidé MeSH
- proteosyntéza * MeSH
- senioři MeSH
- triple-negativní karcinom prsu * etiologie genetika imunologie patofyziologie MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- senioři MeSH
- Publikační typ
- klinická studie MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
- Klíčová slova
- atezolumab, pembrolizumab,
- MeSH
- antigeny CD274 antagonisté a inhibitory MeSH
- bevacizumab MeSH
- geny BRCA1 účinky léků MeSH
- geny BRCA2 účinky léků MeSH
- lidé MeSH
- lokální recidiva nádoru farmakoterapie patologie MeSH
- PARP inhibitory terapeutické užití MeSH
- protokoly antitumorózní kombinované chemoterapie aplikace a dávkování MeSH
- taxoidy terapeutické užití MeSH
- triple-negativní karcinom prsu * epidemiologie farmakoterapie genetika patologie MeSH
- Check Tag
- lidé MeSH
- ženské pohlaví MeSH
- MeSH
- COVID-19 psychologie MeSH
- koordinovaný terapeutický postup normy organizace a řízení MeSH
- lékařská onkologie * metody organizace a řízení MeSH
- lidé MeSH
- triple-negativní karcinom prsu diagnostické zobrazování farmakoterapie genetika MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- novinové články MeSH
- rozhovory MeSH
A growing body of evidence supports the notion that cancer resistance is driven by a small subset of cancer stem cells (CSC), responsible for tumor initiation, growth, and metastasis. Both CSC and chemoresistant cancer cells may share common qualities to activate a series of self-defense mechanisms against chemotherapeutic drugs. Here, we aimed to identify proteins in chemoresistant triple-negative breast cancer (TNBC) cells and corresponding CSC-like spheroid cells that may contribute to their resistance. We have identified several candidate proteins representing the subfamilies of DNA damage response (DDR) system, the ATP-binding cassette, and the 26S proteasome degradation machinery. We have also demonstrated that both cell types exhibit enhanced DDR when compared to corresponding parental counterparts, and identified RAD50 as one of the major contributors in the resistance phenotype. Finally, we have provided evidence that depleting or blocking RAD50 within the Mre11-Rad50-NBS1 (MRN) complex resensitizes CSC and chemoresistant TNBC cells to chemotherapeutic drugs.
- MeSH
- chemorezistence účinky léků genetika MeSH
- cisplatina aplikace a dávkování MeSH
- cyklofosfamid aplikace a dávkování MeSH
- DNA vazebné proteiny genetika MeSH
- doxorubicin aplikace a dávkování MeSH
- enzymy opravy DNA genetika MeSH
- homologní protein MRE11 genetika MeSH
- hydrolasy působící na anhydridy kyselin genetika MeSH
- jaderné proteiny genetika MeSH
- lidé MeSH
- nádorové kmenové buňky účinky léků metabolismus MeSH
- poškození DNA účinky léků MeSH
- přežití po terapii bez příznaků nemoci MeSH
- proteiny buněčného cyklu genetika MeSH
- triple-negativní karcinom prsu farmakoterapie genetika MeSH
- Check Tag
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Triple negative breast cancers (TNBC) are a morphologically and genetically heterogeneous group of breast cancers with uncertain prediction of biological behavior and response to therapy. Epithelial to mesenchymal transition (EMT) is a dynamic process characterized by loss of typical epithelial phenotype and acquisition of mesenchymal characteristics. Aberrant activation of EMT can aggravate the prognosis of patients with cancer, however, the mechanisms of EMT and role of microRNAs (miRNAs) in EMT activation is still unclear. The aim of our study was to analyze miRNA expression within areas of TNBCs with cellular morphology that may be related to the EMT process and discuss possible associations. Out of all 3953 re-examined breast cancers, 460 breast cancers were diagnosed as TNBC (11.64%). With regard to complete tumor morphology preservation, the tissue samples obtained from core-cut biopsies and influenced by previous neoadjuvant therapy were excluded. We assembled a set of selected 25 cases to determine miRNA expression levels in relation to present focal spindle cell and apocrine cell morphology within individual TNBCs. We used descriptive (histological typing and morphology), morphometric, molecular (microdissection of tumor and non-tumor morphologies, RNA isolation and purification, microchip analysis) and bioinformatic analysis (including pathway analysis). The results were verified by quantitative real-time PCR (RT-qPCR) on an extended set of 70 TNBCs. The majority of TNBCs were represented by high-grade invasive carcinomas of no special type (NST) with medullary features characterized by well-circumscribed tumors with central necrosis or fibrosis and frequent tendency to spindle-cell and/or apocrine cell transformation. Apocrine and spindle cell transformation showed a specific miRNA expression profile in comparison to other tumor parts, in situ carcinoma or non-tumor structures, particularly down-regulated expression of hsa-miRNA-143-3p and hsa-miRNA-205-5p and up-regulated expression of hsa-miR-22-3p, hsa-miRNA-185-5p, and hsa-miR-4443. Apocrine cell tumor morphology further revealed decreased expression of hsa-miR-145-5p and increased expression of additional 14 miRNAs (e.g. hsa-miR-182-5p, hsa-miR-3135b and hsa-miR-4417). Pathway analysis for target genes of these miRNAs revealed several shared biological processes (i.e. Wnt signaling, ErbB signaling, MAPK signaling, endocytosis and axon guidance), which may in part contribute to the EMT and tumor progression. We provide the first miRNA expression profiling of specific tissue morphologies in TNBC. Our results demonstrate a specific miRNA expression profile of apocrine and spindle cell morphology which can exhibit a certain similarity with the EMT process and may also be relevant for prognosis and therapy resistance of TNBC.
- MeSH
- apokrinní žlázy * mikrobiologie patologie MeSH
- dospělí MeSH
- epitelo-mezenchymální tranzice * genetika MeSH
- lidé středního věku MeSH
- lidé MeSH
- mikro RNA * genetika MeSH
- regulace genové exprese u nádorů genetika MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- signální dráha Wnt genetika MeSH
- stanovení celkové genové exprese MeSH
- triple-negativní karcinom prsu * genetika patologie MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- ženské pohlaví MeSH
