Endothelial progenitor cells (EPCs) and mesenchymal stem/stromal cells (MSCs) are associated with maintaining tissue homeostasis and tissue repair. Both types of cells contribute to tissue regeneration through the secretion of trophic factors (alone or in the form of microvesicles). This study investigated the isolation and biological properties of microvesicles (MVs) derived from human immortalized MSC line HATMSC1 of adipose tissue origin and EPC line. The human immortalized cell line derived from the adipose tissue of a patient with venous stasis was established in our laboratory using the hTERT and pSV402 plasmids. The human EPC line originating from cord blood (HEPC-CB.1) was established in our previous studies. Microvesicles were isolated through a sequence of centrifugations. Analysis of the protein content of both populations of microvesicles, using the Membrane-Based Antibody Array and Milliplex ELISA showed that isolated microvesicles transported growth factors and pro- and antiangiogenic factors. Analysis of the miRNA content of isolated microvesicles revealed the presence of proangiogenic miRNA (miR-126, miR-296, miR-378, and miR-210) and low expression of antiangiogenic miRNA (miR-221, miR-222, and miR-92a) using real-time RT-PCR with the TaqMan technique. The isolated microvesicles were assessed for their effect on the proliferation and proangiogenic properties of cells involved in tissue repair. It was shown that both HEPC-CB.1- and HATMSC1-derived microvesicles increased the proliferation of human endothelial cells of dermal origin and that this effect was dose-dependent. In contrast, microvesicles had a limited impact on the proliferation of fibroblasts and keratinocytes. Both types of microvesicles improved the proangiogenic properties of human dermal endothelial cells, and this effect was also dose-dependent, as shown in the Matrigel assay. These results confirm the hypothesis that microvesicles of HEPC-CB.1 and HATMSC1 origin carry proteins and miRNAs that support and facilitate angiogenic processes that are important for cutaneous tissue regeneration.

Active cellular transporters of harmful agents-multidrug resistance (mdr) proteins-are present in tumor, stem and endothelial cells, among others. While mdr proteins are broadly studied in tumor cells, their role in non-tumor cells and the significance of their action not connected with removal of harmful xenobiotics is less extensively documented. Proper assessment of mdr proteins expression is difficult. Mdr mRNA presence is most often evaluated but that does not necessarily correlate with the protein level. The protein expression itself is difficult to determine; usually cells with mdr overexpression are studied, not cells under physiological conditions, in which a low expression level of mdr protein is often insufficient for detection in vitro. Various methods are used to identify mdr mRNA and protein expression, together with functional tests demonstrating their biological drug transporting activities. Data comparing different methods of investigating expression of mdr mRNAs and their corresponding proteins are still scarce. In this article we present the results of a study concerning mdr mRNA and protein expression. Our goal was to search for the best method to investigate the expression level and functional activity of five selected mdr proteins-MDR1, BCRP, MRP1, MRP4 and MRP5-in established in vitro cell lines of human endothelial cells (ECs) and their progenitors. Endothelial cells demonstrated mdr presence at the mRNA level, which was not always confirmed at the protein level or in functional tests. Therefore, several different assays had to be applied for evaluation of mdr proteins expression and functions in endothelial cells. Among them functional tests seemed to be the most conclusive, although not very specific.

Angiogenesis is a multi-stage process of new vessel development which involves migration, proliferation and differentiation of endothelial cells. Pathological angiogenesis plays a crucial role in the pathomechanism of various ischemic, malignant and inflammatory disorders. Among eye diseases, macular degeneration (AMD) and proliferative diabetic retinopathy are a major public health issue as the most common causes of blindness. Since angiogenesis plays a crucial role in these conditions, there has been an increased interest in evaluating anti-angiogenic agents in their treatment. The polyphenol resveratrol found in the skin of red grapes, red wine, peanuts and other natural sources, controls proliferation of the cells, induces differentiation and induces apoptosis in various malignant cell lines. Modulation of angiogenesis by this compound has been considered as a very exciting topic and subject of further investigations. The aim of our study was in vitro assessment of resveratrol's influence on proliferation, migration and invasion of an immortalized murine endothelial cell line from peripheral lymph node HEC clone a10. Resveratrol was shown to inhibit the proliferation of the endothelial cells in MTT (at 1, 10 and 50 µM) and AlamarBlue (at 50 µM) assays, and at a concentration of 50 µM significantly inhibited migration of endothelial cells. A concentration-dependent decrease in invasion potential of endothelial cells incubated with resveratrol 10 µM and 50 µM was detected. These promising in vitro results might encourage investigators to test efficacy and safety of resveratrol more extensively in the clinical practice, as a natural and safe anti-angiogenic agent.

Analogs of 1,25-dihydroxyvitamin D3 with a reversed configuration at C-1 or C-24 and E or Z geometry of the double bond at C-22 in the side chain or at C-5 in the triene system were examined for their antiproliferative activity in vitro against a spectrum of various human cancer cell lines. The analogs coded PRI-2201 (calcipotriol), PRI-2202 and PRI-2205, such as calcitriol and tacalcitol (used as a referential agents), revealed antiproliferative activity against human HL-60, HL-60/MX2, MCF-7, T47D, SCC-25 and mouse WEHI-3 cancer cell lines. The toxicity studies in vivo showed that PRI-2202 and PRI-2205 are less toxic than referential agents. Even at total doses of 2.5-5.0 mg/kg distributed during 5 successive days, no changes in body weight were observed. Calcitriol and tacalcitol showed toxicity in the same protocol at 100 times lower doses. Calcipotriol was lethal to all mice after administration of a total dose of 5.0 mg/kg. The analog PRI-2205 appeared to be more active in mouse Levis lung cancer tumor growth inhibition than calcitriol, calcipotriol or PRI-2202. This analog did not reveal calcemic activity at doses which inhibit tumor growth in vivo nor at higher doses.

Bacteriophages (phages) as bacterial viruses are generally believed to have no intrinsic tropism for mammalian cells. In this study the interactions between phages and various eukaryotic cells were investigated. Binding of phages to the membranes of cancer and normal blood cells was observed. Moreover, it was shown that the wild-type phage T4 (wtT4) and its substrain HAP1 with enhanced affinity for melanoma cells inhibit markedly and significantly experimental lung metastasis of murine B16 melanoma cells by 47% and 80%, respectively. A possible molecular mechanism of these effects, namely a specific interaction between the Lys-Gly-Asp motif of the phage protein 24 and beta3-integrin receptors on target cells is proposed. It was also shown that anti-beta3 antibodies and synthetic peptides mimicking natural beta3 ligands inhibit the phage binding to cancer cells. This is in line with the well-described beta3 integrin-dependent mechanism of tumor metastasis. It is concluded that the blocking of beta3 integrins by phage preparations results in a significant decrease in tumor invasiveness.

Synthesis and properties of rhodium complexes with nitrogen ligands [RhCl2(Hpz)4][RhCl4(Hpz)2] (1), [RhCl3(tpy)] (2), [RhCl3(tpta)]·H2O (3) and [Rh(tpy)2(Him)]Cl3·3H2O (4), have been described. X-ray structures of complexes 1–3 have been determined. IR, UV–Vis and 1H NMR spectra of the complexes have been discussed. Cytostatic activity of the complexes against HCV29T tumor cells increases in the series: 1<3<2<4. The cytostatic activity of complex 4 is greater than that of cisplatin. Interaction of the complexes with DNA has been investigated.

An entry from the Cambridge Structural Database, the world’s repository for small molecule crystal structures. The entry contains experimental data from a crystal diffraction study. The deposited dataset for this entry is freely available from the CCDC and typically includes 3D coordinates, cell parameters, space group, experimental conditions and quality measures.

Numerous vitamin D3 analogues have been synthesised in recent years in order to obtain compounds with a favourable biological and therapeutic (antipsoriatic and/or antitumour) activity. Our results showed that pre-treatment for 72 hours of HL-60 human promyelocytic leukaemia cells with calcitriol or its new analogues significantly potentiated their sensitivity to the antiproliferative effect in vitro of cisplatin, doxorubicin or genistein. Moreover, for all cytotoxic agents tested a synergistic antiproliferative effect was observed. This effect was expressed as a significant decrease of the ID50 (inhibitory dose 50%) values for each cytotoxic agent applied after pretreatment with calcitriol or its analogues of HL-60 cells in comparison with the effect of cytotoxic agent applied alone. The observed in vitro potentiated antiproliferative effect of cytotoxic drugs used in combination with vitamin D or its analogues may raise the question as to whether such an effect could be expected in the in vivo situation.

Reactions of [Rh2(µ-OAc)4(H2O)2] ([1·(H2O)2]) with tris(3-methoxyphenyl)phosphine at 1:1 and 1:2 molar ratios yield, first, the appropriate adducts: [1·(H2O){P(C6H4-3-OMe)3}] and [1·{P(C6H4-3-OMe)3}2], and then [Rh2(µ-OAc)3{µ-(C6H3-3-OMe)P(C6H4-3-OMe)2}(HOAc)2] ([2·(HOAc)2]), and [Rh2(µ-OAc)2{µ-(C6H3-3-OMe)P(C6H4-3-OMe)2}2(HOAc)2] ([3·(HOAc)2]) complexes, respectively. They have been characterized by spectroscopic methods. The molecular structure of [3·(HOAc)(H2O)] has been determined crystallographically. The complexes [3·(HOAc)2], [Rh2(µ-OAc)3{µ-(C6H3-4-OMe)P(C6H4-4-OMe)2}(HOAc)2] ([4·(HOAc)2]), and [Rh2(µ-OAc)2{µ-(C6H3-4-OMe)P(C6H4-4-OMe)2}2(HOAc)2] ([5·(HOAc)2]) reversibly react with CO giving mono- and biadducts. Antitumor activity of binuclear rhodium(II) compounds [3·(HOAc)2], [Rh2(µ-OAc)3{µ-(C6H3-2-O)P(C6H3-2-OMe)2}(HOAc)] ([6·(HOAc)]), and [Rh2(µ-OAc)3{µ-(C6H3-6-OMe-2-O)P[(C6H3-2,6-(OMe)2]2}(HOAc)] ([7·(HOAc)]) have been investigated in vitro. The most active agent for investigated tumor lines is complex [6·(HOAc)]. It shows higher activity than cisplatin (cis-[PtCl2(NH3)2]). Antitumor activity decreases in the series: [6·(HOAc)] > [7·(HOAc)] > [3·(HOAc)2]. Activity of all investigated rhodium(II) complexes is higher than that of [1·(H2O)2].Key words: dirhodium(II) complexes, functionalized phosphines, aryl phosphines, ferrocenylmethylphosphines, adducts with CO, antitumor activity, orthometallation reactions.

The results of studies on antiproliferative activity in vitro of nine new platinum(II) complexes against cells of eight human and six murine neoplastic cell lines are described. New complexes with the anionic rest originating from enantiomeric forms of hydroxydicarboxylic malic acid were synthesized to obtain agents with increased water solubility and decreased toxicity. Three compounds, coded 1-3, with ethylenediamine as a neutral ligand, showed cytotoxic activity against 12 out of 14 target cell lines. Their cytotoxic activity was similar or even slightly higher than that of the reference carboplatin. The remaining six compounds, coded 4-9, with 1-alkylimidazole as a neutral ligand, revealed rather low cytotoxic activity, and only against the cells of the human bladder cancer cell line Hu1703He, ovarian cancer cell line OAW-42 and mouse leukemia P388. Most of them appeared to be negative against all other cell lines. No compounds, including reference carboplatin, showed any cytotoxicity against the cells of the T47D human breast cancer cell line or B16F-10 mouse melanoma cell line. The results obtained are in accordance with common opinion, i.e. that the presence of neutral amine ligands with NH groups is required for the cytotoxic activity of platinum complexes. Compounds with a primary amine (ethylenediamine) showed higher cytotoxic activity in vitro than complexes with a tertiary amine (1alkylimidazole).

The antiproliferative in vitro activity of side-chain modified analogues of 1,25-dihydroxyvitamin D3 was examined in order to select compounds with potential antitumour activity. Analogues PRI-1906, PRI-1907, PRI-1909, PRI-2191, PRI-2192, PRI-2193 and PRI-2194 were examined for their antiproliferative activity in vitro against a spectrum of various human cancer cell lines using the MTT technique. In addition, analogues PRI-1906 and PRI-2191 were screened against cells of human leukaemia HL-60 line and against normal human skin fibroblasts. Calcitriol and these two analogues revealed strong antiproliferative activity against these two targets with maximal growth inhibition of 68% for HL-60 cells and of 60% for fibroblasts, and this effect was dose dependent. All analogues tested, except PRI-1909, revealed antiproliferative activity against human carcinoma cell lines of breast origin applied, namely against T47D and MCF-7. The maximal growth inhibition of 49% for T47D cell line and 39% for MCF-7 line was observed, and this effect was dose dependent. The inhibitory doses of the analogues tested were compared with the indices for calcitriol. Analogue PRI-1906 revealed the strongest antiproliferative activity against these four target cell lines (HL-60, fibroblasts, MCF-7, and T47D). The novel analogues of calcitriol, similarly to calcitriol, appeared to be not active against other human cancer cell lines tested (including those originated from lung, colon, prostate, urinary bladder, ovary, pancreas, stomach and kidney) revealing an antiproliferative activity not exceeding 20%. The mechanism of the observed antiproliferative effect of calcitriol and its analogues in vitro remains unclear, however, it may be related to their effect on cell differentiation. The appearance of antigen CD14 and CD11b expression after exposure to calcitriol and its new analogues confirmed their effect on cell differentiation.

Four families of human in vitro cell lines were tested for minisatellite restriction fragment length polymorphism (RFLP) using multilocus probes MZ1.3 and/or 33.15 after digestion of DNA with restriction enzymes HinfI or HaeIII. These results confirmed that (i) the RFLP pattern is relatively stable in established cell lines and, therefore, could be used as a specific marker of a cell line identity, (ii) the use of MZ1.3 and 33.15 probes permits the identification of hybridomas and (iii) one of the cell lines tested, a lymphoblastoid cell line HAJ, may possess a hot spot of mutation.

Cytostatic activity of newly synthesized aziridinyl substituted cyclophosphazenes was tested in two in vitro systems. First, the compounds were tested by total cell protein inhibition test on human KB tumor cell line. Selected active compounds were tested in a clonogenic assay on murine L1210 leukemia cells. Out of 18 compounds tested, 12 were newly synthesized, 3 were their substrates and 4 were compounds with known cytostatic activity, used as a positive controls. Four of the newly synthesized compounds, designated with symbols: BANF-4Az, BBNF-4Az. T-oFDA-Az and W-10 revealed cytostatic activity for KB cells (ED50 = 3.4-19 micrograms/mL) and were tested in clonogenic assay. One of these compounds, namely T-oFDA-Az was subsequently selected for the further steps of screening for antitumor activity in animal tumor models.

Cytostatic activity of 48 new amino acid analogs and derivatives of phosphonic acid type was tested against KB cell line in vitro. The tests were performed according to the recommended international protocol for screening of chemical agents against tissue culture system. Three of the compounds tested: No. No, 29, 33 and 32 revealed cytostatic activity at the concentrations: 63, 150, and 180 micrograms ml-1 respectively (ID50).