Chelidonin a Homochelidonin indukují buněčnou smrt prostřednictvím drah checkpoint a MAP kináz

This study focuses on the comparative in vitro cytotoxicity of chelidonine and homochelidonine on human cancer and non-cancer cells. Both alkaloids produced a decrease in cellular growth in a dose-dependent manner exhibiting greater potency in cancer cells. The growth inhibitory effect was evidenced in both ovarian carcinoma A2780 and lung fibroblast MRC-5 cells by inducing G2 and mitotic phase cell cycle arrest. Results indicated that the extent of apoptosis induced by chelidonine and homochelidonine was correlated to sensitivity to the antiproliferative activity of the evaluated compounds. Western blotting suggested that the cellular toxicological mechanism of chelidonine is related to the differential upregulation of phospho-Chk2, p21(Cip1/Waf1), phospho-ERK1/2 and phospho-p38 in various cell types, leading to alternations in the suppression of proliferation and either induction or prevention of apoptosis. Chelidonine showed the more potent effects and also affected the cell cycle checkpoints and MAPK signaling pathways within cells.Tato práce je zaměřena na srovnávací cytotoxicitu chelidoninu a homochelidoninu na lidských nádorových a nenádorových buňkách

Pankracin, alkaloid čeledi Amaryllidaceae montaninového typu, inhibuje proliferaci buněk plicního adenokarcinomu A549 a indukuje apoptózu u leukemických buněk MOLT-4

Pancracine, a montanine-type Amaryllidaceae alkaloid (AA), is one of the most potent compounds among natural isoquinolines. In previous studies, pancracine exhibited cytotoxic activity against diverse human cancer cell lines in vitro. However, further insight into the molecular mechanisms that underlie the cytotoxic effect of pancracine have not been reported and remain unknown. To fill this void, the cell proliferation and viability of cancer cells was explored using the Trypan Blue assay or by using the xCELLigence system. The impact on the cell cycle was determined by flow cytometry. Apoptosis was evaluated by Annexin V/PI and by quantifying the activity of caspases (-3/7, -8, and -9). Proteins triggering growth arrest or apoptosis were detected by Western blotting. Pancracine has strong antiproliferative activity on A549 cells, lasting up to 96 h, and antiproliferative and cytotoxic effects on MOLT-4 cells. The apoptosis-inducing activity of pancracine in MOLT-4 cells was evidenced by the significantly higher activity of caspases. This was transmitted through the upregulation of p53 phosphorylated on Ser392, p38 MAPK phosphorylated on Thr180/Tyr182, and upregulation of p27. The pancracine treatment negatively altered the proliferation of A549 cells as a consequence of an increase in G1-phase accumulation, associated with the downregulation of Rb phosphorylated on Ser807/811 and with the concomitant upregulation of p27 and downregulation of Akt phosphorylated on Thr308. This was the first study to glean a deeper mechanistic understanding of pancracine activity in vitro. Perturbation of the cell cycle and induction of apoptotic cell death were considered key mechanisms of pancracine action.Pankracin, alkaloid čeledi Amaryllidaceae (AA) montaninového typu, je jednou z nejúčinnějších sloučenin mezi přírodními isochinoliny. V předchozích studiích pankracin vykazoval cytotoxickou aktivitu proti různým lidským nádorovým buněčným liniím. Nicméně, detailní molekulární mechanismus, cytotoxického účinku pankracinu, však nebyl popsán a zůstává neznámý. K vyplnění této mezery byla zkoumána buněčná proliferace a viabilita nádorových buněk pomocí testu vylučování trypanové modře nebo pomocí systému xCELLigence. Vliv na buněčný cyklus byl stanoven pomocí průtokové cytometrie. Apoptóza byla hodnocena pomocí Annexinu V/PI a kvantifikací aktivity kaspáz (-3/7, -8 a -9). Proteiny spouštějící zástavu buněčného cyklu a apoptózu byly detekovány pomocí Western blotu. Pankracin má silnou antiproliferativní aktivitu na buňky A549, která trvá až 96 hodin, a antiproliferativní a cytotoxické účinky na buňky MOLT-4. Apoptózu indukující aktivita pankracinu u buněk MOLT-4 byla prokázána významně vyšší aktivitou kaspáz. Tato aktivita byla zprostředkována prostřednictvím zvýšené exprese p53 fosforylovaného na Ser392, p38 MAPK fosforylovaného na Thr180/Tyr182 a zvýšené exprese p27. Pankracin negativně ovlivnil proliferaci buněk A549 v důsledku zvýšené akumulace buněk v G1 fázi, spojené se sníženou expresí Rb fosforylovaného na Ser807/811 a se současnou zvýšenou expresí p27 a sníženou expresí Akt fosforylovaného na Thr308. Toto je první studie, která shromáždila hlubší poznatky o mechanismu působení pankracinu in vitro. Ovlivnění buněčného cyklu a indukce apoptózy jsou považovány za klíčové mechanismy působení pankracinu

Scoulerine affects microtubule structure, inhibits proliferation, arrests cell cycle and thus culminates in the apoptotic death of cancer cells

Abstract Scoulerine is an isoquinoline alkaloid, which indicated promising suppression of cancer cells growth. However, the mode of action (MOA) remained unclear. Cytotoxic and antiproliferative properties were determined in this study. Scoulerine reduces the mitochondrial dehydrogenases activity of the evaluated leukemic cells with IC50 values ranging from 2.7 to 6.5 µM. The xCELLigence system revealed that scoulerine exerted potent antiproliferative activity in lung, ovarian and breast carcinoma cell lines. Jurkat and MOLT-4 leukemic cells treated with scoulerine were decreased in proliferation and viability. Scoulerine acted to inhibit proliferation through inducing G2 or M-phase cell cycle arrest, which correlates well with the observed breakdown of the microtubule network, increased Chk1 Ser345, Chk2 Thr68 and mitotic H3 Ser10 phosphorylation. Scoulerine was able to activate apoptosis, as determined by p53 upregulation, increase caspase activity, Annexin V and TUNEL labeling. Results highlight the potent antiproliferative and proapoptotic function of scoulerine in cancer cells caused by its ability to interfere with the microtubule elements of the cytoskeleton, checkpoint kinase signaling and p53 proteins. This is the first study of the mechanism of scoulerine at cellular and molecular level. Scoulerine is a potent antimitotic compound and that it merits further investigation as an anticancer drug

Cytotoxicita přirozeně se vyskytujících isochinolinových alkaloidů různých strukturních typů

Forty-six isoquinoline alkaloids, of eleven structural types isolated in our laboratory, have been evaluated for their cytotoxicity against two cancer cell lines (Caco-2 and Hep-G2 cancer cells), as well as against normal human lung fibroblast cells. Only scoulerine, aromoline, berbamine and parfumidine showed significant cytotoxic effects, but only scoulerine was active against both Caco-2 and Hep-G2 cells (IC50 values 6.44 +/- 0.87 and 4.57 +/- 0.42, respectively). Unfortunately, except for parfumidine, the other active alkaloids were also cytotoxic to the normal human lung fibroblast cells.Tato práce popisuje cytotoxicitu čtyřiceti šesti přirozeně se vyskytujících isochinolinových alkaloidů vůči nádorovým buněčným liniím a normálním lidským fibroblastům

Bersavin: Nový bisbenzylisochinolinový alkaloid s cytotoxickými, antiproliferativními a apoptózovými účinky na lidské leukemické buňky

Bersavine is the new bisbenzylisoquinoline alkaloid isolated from the Berberis vulgaris L. (Berberidaceae) plant. The results of cytotoxicity screening 48 h post-treatment showed that bersavine considerably inhibits the proliferation and viability of leukemic (Jurkat, MOLT-4), colon (HT-29), cervix (HeLa) and breast (MCF-7) cancer cells with IC50 values ranging from 8.1 to 11 mu M. The viability and proliferation of leukemic Jurkat and MOLT-4 cells were decreased after bersavine treatment in a time- and dose-dependent manner. Bersavine manifested concentration-dependent antiproliferative activity in human lung, breast, ovarian and hepatocellular carcinoma cell lines using a xCELLigence assay. Significantly higher percentages of MOLT-4 cells exposed to bersavine at 20 mu M for 24 h were arrested in the G1 phase of the cell cycle using the flow cytometry method. The higher percentage of apoptotic cells was measured after 24 h of bersavine treatment. The upregulation of p53 phosphorylated on Ser392 was detected during the progression of MOLT-4 cell apoptosis. Mechanistically, bersavine-induced apoptosis is an effect of increased activity of caspases, while reduced proliferation seems dependent on increased Chk1 Ser345 phosphorylation and decreased Rb Ser807/811 phosphorylation in human leukemic cells.Bersavin je nový bisbenzylisochinolinový alkaloid izolovaný z rostliny Berberis vulgaris L. (Berberidaceae). Výsledky cytotoxického screeningu po 48 hodinách ukázaly, že bersavin významně inhibuje proliferaci a životaschopnost leukemických (Jurkat, MOLT-4), tlustého střeva (HT-29), děložního čípku (HeLa) a karcinomu prsu (MCF-7) s hodnotami IC50 v rozmezí od 8,1 do 11 uM. Viabilita a proliferace leukemických buněk Jurkat a MOLT-4 byla po ovlivnění bersavinem snížena v závislosti na čase a dávce. Bersavin vykazoval na koncentraci závislou antiproliferativní aktivitu u lidských nádorových linií plic, prsu, vaječníků a hepatocelulárního karcinomu pomocí testu xCELLigence. Významně vyšší procento buněk MOLT-4 ovlivněných bersavinem v dávce 20 uM po dobu 24 hodin bylo zastaveno ve fázi G1 buněčného cyklu měřeného pomocí průtokové cytometrie. Vyšší procento apoptotických buněk bylo zjištěno za 24 hodin po ovlivnění bersavinem. Zvýšená exprese p53 fosforylovaného na Ser392 byla detekována během progrese apoptózy u buněk MOLT-4. Apoptóza indukovaná bersavinem je důsledkem zvýšené aktivity kaspáz, zatímco, snížená proliferace závisí pravděpodobně na zvýšené fosforylaci Chk1 Ser345 a snížené fosforylaci Rb Ser807 / 811 u lidských leukemických buněk

Protinádorový potenciál alkaloidů čeledi Amaryllidaceae testovaný na panelu lidských linií, pomocí analýzy buněk v reálném čase a na Ehrlichově nádoru myší

In this study, twenty-two Amaryllidaceae alkaloids were screened for their anticancer potential. All isolates were evaluated for antiproliferative activities on a panel of 17 human cell types of different tissue origin using WST-1 assay. In addition, we determined the antiproliferative effect with a real-time cell analysis xCELLigence system. Thereafter, to evaluate the barely known in vivo anticancer potential of the most potent molecule haemanthamine, a preliminary study was performed using an Ehrlich tumor-bearing mice model. The results showed that haemanthamine, lycorine and haemanthidine exerted the highest antiproliferative activity. The mean growth percent (GP) value after a single-dose 10 mu M treatment was for haemanthamine 21%, for lycorine 21% and for haemanthidine 27% that of untreated control cells (100%). Furthermore, haemanthamine, lycorine and haemanthidine exhibited significant cytotoxicities against all the tested cell lines with individual IC50 values in the micromolar range. Dynamic real-time measures of impedance by xCELLigence indicated that these three compounds suppress cell proliferation after 10 h of treatment at a concentration of 10 mM or higher. Regrettably, in a follow-up in vivo antitumor activity study, haemanthamine showed no statistically significant reduction in the tumor size with no prolongation of survival time of Ehrlich tumor-bearing mice. Taken together, these results provide a new clue and guidance for exploiting Amaryllidaceae alkaloids as anticancer agents.V této studii byl testován protinádorový potenciál 22 alkaloidů čeledi Amaryllidaceae na panelu lidských linií, pomocí analýzy buněk v reálném čase a na Ehrlichově nádoru myší

Porovnání cytotoxicity chelidoninu a homochelidoninu, dimethoxy analoga izolovaného z Chelidonium majus L. (Papaveraceae), vůči lidským leukemickým a plicním nádorovým buňkám

In this study the effects of naturally occurring homochelidonine in comparison to chelidonine on cell cycle progression and cell death in leukemic T-cells with different p53 status are described. We found that homochelidonine and chelidonine displayed significant cytotoxicity in examined blood cancer cells with the exception of HEL 92.1.7 and U-937 exposed to homochelidonine. Unexpectedly, homochelidonine and chelidonine-induced cytotoxicity was more pronounced in Jurkat cells contrary to MOLT-4 cells. Homochelidonine showed an antiproliferative effect on A549 cells but it was less effective compared to chelidonine. Biphasic dose-depended G1 and G2/M cell cycle arrest along with the population of sub-G1 was found after treatment with homochelidonine in MOLT-4 cells. In variance thereto, an increase in G2/M cells was detected after treatment with homochelidonine in Jurkat cells. Treatment with chelidonine induced cell cycle arrest in the G2/M cell cycle in both MOLT-4 and Jurkat cells. MOLT-4 and Jurkat cells treated with homochelidonine and chelidonine showed features of apoptosis such as phosphatidylserine exposure, a loss of mitochondrial membrane potential and an increase in the caspases -3/7, -8 and -9. Western blots indicate that homochelidonine and chelidonine exposure activates Chk1 and Chk2. Studies conducted with fluorescence microscopy demonstrated that chelidonine and homochelidonine inhibit tubulin polymerization in A549 cells. Collectively, the data indicate that chelidonine and homochelidonine are potent inducers of cell death in cancer cell lines, highlighting their potential relevance in leukemic cells.Tato práce porovnává cytotoxicitu chelidoninu a homochelidoninu vůči lidským leukemickým a plicním nádorovým buňkám. Výsledky studie naznačují, že chelidonin a homochelidonin jsou účinnými aktivátory apoptózy u nádorových buněčných linií

Deriváty beta-Crinane Amaryllidaceae Alkaloid Haemanthamine jako vícecílové cílené ligandy pro Alzheimerovu chorobu

Twelve derivatives 1a-1m of the beta-crinane-type alkaloid haemanthamine were developed. All the semisynthetic derivatives were studied for their inhibitory potential against both acetylcholinesterase and butyrylcholinesterase. In addition, glycogen synthase kinase 3 beta (GSK-3 beta) inhibition potency was evaluated in the active derivatives. In order to reveal the availability of the drugs to the CNS, we elucidated the potential of selected derivatives to penetrate through the blood-brain barrier (BBB). Two compounds, namely 11-O-(2-methylbenzoyl)-haemanthamine (1j) and 11-O-(4-nitrobenzoyl)-haemanthamine (1m), revealed the most intriguing profile, both being acetylcholinesterase (hAChE) inhibitors on a micromolar scale, with GSK-3 beta inhibition properties, and predicted permeation through the BBB. In vitro data were further corroborated by detailed inspection of the compounds' plausible binding modes in the active sites of hAChE and hBuChE, which led us to provide the structural determinants responsible for the activity towards these enzymes.Bylo vyvinuto dvanáct derivátů la-lm alkaloidního hematoaminu beta-krinanu. Všechny semisyntetické deriváty byly studovány na inhibiční potenciál proti acetylcholinesteráze i butyrylcholinesteráze. Kromě toho byla v aktivních derivátech hodnocena inhibiční schopnost glykogen syntázy kinázy 3 beta (GSK-3 beta). Abychom odhalili dostupnost léčiv pro CNS, objasnili jsme potenciál vybraných derivátů proniknout hematoencefalickou bariérou (BBB). Dvě sloučeniny, jmenovitě 11-O- (2-methylbenzoyl) -haemanthamin (1j) a 11-O- (4-nitrobenzoyl) -haemanthamin (1m), odhalily nejzajímavější profil, obě jsou inhibitory acetylcholinesterázy (hAChE) na mikromolárním v měřítku, s inhibičními vlastnostmi GSK-3 beta a předpovězenou permeací BBB. Data in vitro byla dále potvrzena podrobnou inspekcí věrohodných vazebných režimů sloučenin v aktivních místech hAChE a hBuChE, což nás vedlo k poskytnutí strukturálních determinant odpovědných za aktivitu vůči těmto enzymům

Flavones Inhibit the Activity of AKR1B10, a Promising Therapeutic Target for Cancer Treatment

AKR1B10 is an NADPH-dependent reductase that plays an important function in several physiological reactions such as the conversion of retinal to retinol, reduction of isoprenyl aldehydes, and biotransformation of procarcinogens and drugs. A growing body of evidence points to the important role of the enzyme in the development of several types of cancer (e.g., breast, hepatocellular), in which it is highly overexpressed. AKR1B10 is regarded as a therapeutic target for the treatment of these diseases, and potent and specific inhibitors may be promising therapeutic agents. Several inhibitors of AKR1B10 have been described, but the area of natural plant products has been investigated sparingly. In the present study almost 40 diverse phenolic compounds and alkaloids were examined for their ability to inhibit the recombinant AKR1B10 enzyme. The most potent inhibitorsapigenin, luteolin, and 7-hydroxyflavonewere further characterized in terms of IC₅₀, selectivity, and mode of action. Molecular docking studies were also conducted, which identified putative binding residues important for the interaction. In addition, cellular studies demonstrated a significant inhibition of the AKR1B10-mediated reduction of daunorubicin in intact cells by these inhibitors without a considerable cytotoxic effect. Although these compounds are moderately potent and selective inhibitors of AKR1B10, they constitute a new structural type of AKR1B10 inhibitor and may serve as a template for the development of better inhibitors

Antiproliferative activity and apoptosis-inducing mechanism of Amaryllidaceae alkaloid montanine on A549 and MOLT-4 human cancer cells

The isoquinoline alkaloids found in Amaryllidaceae are attracting attention due to attributes that can be harnessed for the development of new drugs. The possible molecular mechanisms by which montanine exerts its inhibitory effects against cancer cells have not been documented. In the present study, montanine, manthine and a series of 15 semisynthetic montanine analogues originating from the parent alkaloid montanine were screened at a single test dose of 10 μM to explore their cytotoxic activities against a panel of eight cancer cell lines and one non-cancer cell line. Among montanine and its analogues, montanine and its derivatives 12 and 14 showed the highest cytostatic activity in the initial single-dose screening. However, the native montanine exhibited the greatest antiproliferative activity against cancer cells, with a lower mean IC50 value of 1.39 µM, compared to the displayed mean IC50 values of 2.08 µM for 12 and 3.57 µM for 14. Montanine exhibited the most potent antiproliferative activity with IC50 values of 1.04 µM and 1.09 µM against Jurkat and A549 cell lines, respectively. We also evaluated montanine’s cytotoxicity and cell death mechanisms. Our results revealed that montanine triggered apoptosis of MOLT-4 cells via caspase activation, mitochondrial depolarisation and Annexin V/PI double staining. The Western blot results of MOLT-4 cells showed that the protein levels of phosphorylated Chk1 Ser345 were upregulated with increased montanine concentrations. Our findings provide new insights into the mechanisms underlying the cytostatic, cytotoxic and pro-apoptotic activities of montanine alkaloids in lung adenocarcinoma A549 and leukemic MOLT-4 cancer cell types