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EN
We have demonstrated for the first time that the steroid metabolite, 2-methoxyestradiol (2-ME) is a powerful growth inhibitor of human osteosarcoma 143 B cell line by pleiotropic mechanisms involving cell cycle arrest at two different points and apoptosis. The ability of 2-ME to inhibit cell cycle at the respective points has been found concentration dependent. 1 μM 2-ME inhibited cell cycle at G1 phase while 10 μM 2-ME caused G2/M cell cycle arrest. As a natural estrogen metabolite 2-ME is expected to perturb the stability of microtubules (MT) in vivo analogously to Taxol - the MT binding anticancer agent. Contrary to 2-ME, Taxol induced accumulation of osteosarcoma cells in G2/M phase of cell cycle only. The presented data strongly suggest two different mechanisms of cytotoxic action of 2-ME at the level of a single cell.
EN
Camptothecin (CPT), an alkaloid natural product, extracted from Camptotheca acuminata bark, has been reported to have potential antitumor activity in diverse cancers. MicroRNAs (MiRNAs) are a class of short, non-coding RNAs that plays a crucial role in the normal physiology by attenuating translation. Here, we showed that the CPT modulates the expression of miRNAs in hepatocellular carcinoma cells (HCC). Microarray analysis reveals that CPT modulates the expression of as many as 39 miRNAs in HCC cells (Huh7), 27 miRNAs were downregulated whereas 12 miRNAs were upregulated. miR-16 is the key miRNA upregulated by CPT and targets key prosurvival proteins (MMP-2, MMP-9 and cyclin D1). Our results demonstrate that CPT is inhibiting cell viability of HCC cells significantly when compared with the untreated cells. Wound healing and colony formation assay confirm inhibition of cell migration and clonogenic property of Huh7 cells respectively, upon the dose-dependent treatment of CPT. Furthermore, the Boyden chamber assay analysis revealed a significant inhibition of number of invasive cells in CPT treated cells with comparison to untreated Huh7 cells. Mechanistically, CPT upregulates miR-16 expression which targets MMP-2, MMP-9, cyclin D1 downregulation and subsequently upregulates the expression of E-cadherin, TIMP1, p21, and p27, thereby inhibits cell migration, invasion and clonogenic property of HCC cells. In summary, CPT treatment in Huh7 cells decreases cell viability and upregulates miR-16 expression, which results in inhibition of cell migration, invasion and clonogenic property of cells, by decreasing MMP-2, MMP-9, cyclin D1 and increasing the expression of cell cycle-regulated proteins p21 and p27.
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2017
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vol. 64
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issue 2
371-374
EN
ABCC10 (MRP7) plays a role in cellular detoxification and resistance to anticancer drugs. Since ABCC10 gene transcription in human prostate cancer CWR22Rv1 cells was found dependent on E2F binding sequence motif, ABCC10 expression in G1 and S phases of the cell cycle of CWR22Rv1 cells, was analyzed. The cells were synchronized in G1 phase by double thymidine block and in S phase by thymidine/mimosine double block. ABCC10 mRNA level was found to be similar in S phase-synchronized and asynchronous cell populations. In G1 phase it decreased by 2.4- to 3-fold. It is thus inferred, that ABCC10 expression in CWR22Rv1 cells is not S phase-specific but is primarily associated with cell proliferation.
EN
The aim of this study was to expand our knowledge about anticancer activity of some polyamine derivatives with quinoline or chromane as terminal moieties. Tested compounds were evaluated in vitro towards metastatic human prostate adenocarcinoma (PC3), human carcinoma (DU145) and mammary gland adenocarcinoma (MCF7) cell lines. Cell viability was estimated on the basis of mitochondrial metabolic activity using water-soluble tetrazolium WST1 to establish effective concentrations of the tested compounds under experimental conditions. Cytotoxic potential of polyamine derivatives was determined by the measurement of lactate dehydrogenase activity released from damaged cells, changes in mitochondrial membrane potential, the cell cycle distribution analysis and apoptosis assay. It was revealed that the tested polyamine derivatives differed markedly in their antiproliferative activity. Bischromane derivative 5a exhibited a rather cytostatic than cytotoxic effect on the tested cells, whereas quinoline derivative 3a caused changes in cell membrane integrity, inhibited cell cycle progression, as well as induced apoptosis of prostate and breast cancer cells which suggest its potential application in cancer therapy.
EN
The human SUV3 helicase (SUV3, hSUV3, SUPV3L1) is a DNA/RNA unwinding enzyme belonging to the class of DexH-box helicases. It localizes predominantly in the mitochondria, where it forms an RNA-degrading complex called mitochondrial degradosome with exonuclease PNP (polynucleotide phosphorylase). Association of this complex with the polyA polymerase can modulate mitochondrial polyA tails. Silencing of the SUV3 gene was shown to inhibit the cell cycle and to induce apoptosis in human cell lines. However, since small amounts of the SUV3 helicase were found in the cell nuclei, it was not clear whether the observed phenotypes of SUV3 depletion were of mitochondrial or nuclear origin. In order to answer this question we have designed gene constructs able to inhibit the SUV3 activity exclusively in the cell nuclei. The results indicate that the observed growth rate impairment upon SUV3 depletion is due to its nuclear function(s). Unexpectedly, overexpression of the nuclear-targeted wild-type copies of the SUV3 gene resulted in a higher growth rate. In addition, we demonstrate that the SUV3 helicase can be found in the HeLa cell nucleoli, but it is not detectable in the DNA-repair foci. Our results indicate that the nucleolar-associated human SUV3 protein is an important factor in regulation of the cell cycle.
EN
Non-small cell lung cancer (NSCLC) accounts for the majority (85%) of all lung cancers. Although many therapies are available, 35–50% of patients with stage I or II NSCLC develop recurrence and metastasis. This study was designed to investigate the anti-tumor activity of cyanidin (Cy) and peonidin (Pn) on NSCLC cells (SPCA-1). SPCA-1 cell proliferation, cell cycle and early apoptosis were investigated after treatment with Cy and Pn. The underlying signaling mechanism was also explored by detecting the levels of apoptosis-related proteins using enzyme-linked immunosorbent assay (ELISA). Cy and Pn inhibited the viability of SPCA-1 cells with an IC50 of 141.08 μg/mL and 161.31 μg/mL, respectively. Meanwhile, Cy and Pn induced cell cycle arrest at G2/M phase. Cy and Pn treatment significantly increased the levels of Bax, P53, and Caspase-3, while decreasing that of Bcl-2, thereby inhibiting the growth of SPCA-1 cells. In conclusion, Cy and Pn induced early apoptosis of NSCLC cells through regulation of the levels on Caspase-3, Bax, Bcl-2, and P53. These results suggest Cy and Pn as potential anticancer drugs for the treatment of lung cancer.
EN
Protein phosphatase 2A (PP2A) comprises a diverse family of phosphoserine-and phosphothreonine-specific phosphatases present in all eukaryotic cells. All forms of PP2A contain a catalytic subunit (PP2Ac) which forms a stable complex with the structural subunit PR65/A. The heterodimer PP2Ac-PR65/A associates with regulatory proteins, termed variable subunits, in order to form trimeric holoenzymes attributed with distinct substrate specificity and targeted to different subcellular compartments. PP2Ac activity can be modulated by reversible phosphorylation on Tyr307 and methylation on C-terminal Leu309. Studies on PP2A have shown that this enzyme may be implicated in the regulation of metabolism, transcription, RNA splicing, translation, differentiation, cell cycle, oncogenic transformation and signal transduction.
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Biogeneza rzęski pierwotnej

63%
PL
Rzęski pierwotne, struktury zbudowane na bazie cytoszkieletu mikrotubularnego, występują na powierzchni niemal wszystkich komórek ssaczych. Dzięki licznym receptorom błonowym, rzęski pierwotne pośredniczą w odbieraniu i przekazywaniu bodźców ze środowiska do wnętrza komórki, i tym samym odgrywają niezwykle ważną rolę w prawidłowym rozwoju i funkcjonowaniu większości tkanek i narządów. Tworzenie rzęski (ciliogeneza) to złożony, wieloetapowy i wielopoziomowo regulowany proces ściśle związany z cyklem komórkowym. Mutacje w genach kodujących białka strukturalne lub odpowiedzialne za prawidłowe funkcjonowanie rzęsek, jak również, regulujące przebieg ciliogenezy są przyczyną ich dysfunkcji, prowadzącej w efekcie do wielonarządowych chorób zwanych ciliopatiami.
EN
Cilia are highly specialized, microtubule-based protrusions, extended on cell surface in almost all mammalian cell types. They function as cell antennae that receive and transmit signals from the environment to the cell body. Cilia formation, so-called ciliogenesis is strictly controlled at multiple levels by a number of proteins, and correlated with the cell cycle progression. Cilia dysfunctions cause a wide range of human diseases, called ciliopathies. Moreover, ciliary defects may lead to obesity and cancer. In this article, we summarize current knowledge concerning cilia function and structure, regulation of ciliogenesis, and the most important signaling pathways and diseases affected by cilia dysfunction.
EN
O-GlcNAcylation is a post-translational modification involving the addition of a N-acetylglucosamine moiety to the serine/threonine residues of cytosolic or nuclear proteins. Two enzymes are responsible for cyclic O-GlcNAcylation: O-GlcNAc transferase (OGT) which catalyzes the addition of the GlcNAc moiety from UDP-GlcNAc to target proteins and O-GlcNAcase (OGA) which catalyses the hydrolytic removal of the sugar moiety from proteins. Dynamic and reversible O-GlcNAcylation is emerging as an important regulator of diverse cellular processes, such as signal transduction, metabolism, transcription, translation, proteasomal degradation and cell cycle. O-GlcNAcylation occurs on serine or threonine residues of proteins at sites that may also be phosphorylated. Therefore, an extensive crosstalk exists between phosphorylation and O-GlcNAcylation. Recent studies indicate that increased O-GlcNAcylation is a general feature of cancer. Elevated O-GlcNAcylation (hyper-OGlcNAcylation) occurs in many human malignancies including solid tumors such as lung, prostate, breast, colorectal, liver, pancreatic cancers as well as non-solid cancers such as chronic lymphocytic leukemia. The changes in O-GlcNAcylation are associated with the changes in OGT and OGA expression levels. Hyper-O-GlcNAcylation may be linked to the various hallmarks of cancer, including cancer cell proliferation, survival, invasion, metastasis and metabolism. This paper reviews recent findings related to O-GlcNAc-dependent regulation of signaling pathways, cell cycle, transcription factors, and metabolic enzymes in cancer cells.
PL
O-GlcNAcylacja jest odwracalną potranslacyjną modyfikacją białek polegającą na przyłączeniu wiązaniem O-glikozydowym pojedynczych reszt β-N-acetyloglukozaminy (GlcNAc) do seryny lub treoniny. W proces O-GlcNAcylacji włączone są dwa enzymy: O-GlcNAc transferaza (OGT), enzym odpowiedzialny za przyłączanie reszt N-acetyloglukozaminy i β-N-acetyloglukozaminidaza (OGA), która katalizuje reakcję odłączania reszt GlcNAc. Dynamiczna i odwracalna O-GlcNAcylacja odgrywa istoną rolę w regulacji szeregu procesów komórkowych, takich jak przekazywanie sygnału, metabolizm, transkrypcja, translacja, degradacja białek w proteasomach i cykl komórkowy. Ponieważ O-GlcNAcylacja dotyczy reszt seryny lub treoniny, które znajdują się w miejscach rozpoznawanych przez kinazy białkowe, wpływa ona na poziom fosforylacji wielu białek i isnieje ścisła zależność pomiędzy tymi modyfikacjami. Ostanie badania wskazują, że w komórkach nowotworowych dochodzi do znacznego zwiększenia poziomu O-GlcNAcylacji. Hiper-O-GlcNAcylację stwierdzono w różnych typach nowotworów, włączając w to guzy lite np. płuc, prostaty, piersi, jelita grubego, trzustki, wątroby a także białaczki np. przewlekłą białaczkę limfatyczną. Zaburzenia O-GlcNAcylacji związane są ze zmianami w komórkach nowotworowych ekspresji enzymów odpowiedzialnych za ten proces, tj. OGT i OGA. Hiper-O-GlcNAcylacja wpływa na proliferację, przeżycie i metabolizm komórek nowotworowych, jak również zwiększa ich zdolność do inwazji i metastazy. Prezentowana praca stanowi przegląd aktualnych informacji dotyczących roli O-GlcNAcylacji w regulacji szlaków przekazywania sygnałów, cyklu komórkowego, czynników transkrypcyjnych oraz enzymów i innych białek związanych z metabolizmem komórek nowotworowych.
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