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.
Long-term stability of retention times of a wide range of analytes has been evaluated using eight different stationary phases. These were from a single manufacturer to minimize the differences in silanol activity caused by the manufacturing process. The tested stationary phases included bridge ethylene hybrid, 2-ethylpyridine bridge ethylene hybrid with direct modification of silica particles, bidentate crosslinked charged surface hybrid fluorophenyl, bidentate crosslinked high strength silica C18, and propanediol linked phases including diol (pure propanediol linker), and three phases based on diol further modified with 2-picolylamine, diethylamine, and 1-aminoanthracene group. Retention times were monitored at the first injection, after three, nine, twelve months, and after the column regeneration via washing with pure water. The analyses were carried out using three different mobile phases, including methanol, methanol with 10 mmol/L ammonium formate, and methanol with 0.1% ammonium hydroxide. No overall decreasing or increasing trends were observed after evaluating individual contributing parameters such as analyte, stationary phase, and organic modifier. Our results suggest that the silyl-ether formation is not the only factor contributing to changes in the stationary phase pore surface. Indeed, the adsorption of mobile phase additives is probably another significant factor. That was also confirmed by the regeneration procedure using water, which is likely to reverse the silyl-ether formation to achieve the original retention. However, the retention times returned to the original values for all analytes only on three columns. Retention times on other columns remained shifted within ± 15 % RSD depending on the analyte properties and the nature of organic modifier. The retention time variations observed for each analyte group, i.e., acids, bases, and neutrals, were interpreted for each stationary phase. We concluded that the sterically protected surfaces exhibited significantly smaller changes in the retention times. Although the regeneration procedure effect depended on the column type, the results suggested beneficial effect of water. However, as the adsorption of additives on the column surface is an additional factor leading to retention time variations, the recommendation of using only one additive and/or organic modifier in each column will clearly improve the long-term repeatability of the retention times.
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.
- MeSH
- aktivace enzymů účinky léků MeSH
- antitumorózní látky chemie farmakologie MeSH
- apoptóza účinky léků MeSH
- berberinové alkaloidy chemie farmakologie MeSH
- estery chemie MeSH
- fosforylace účinky léků MeSH
- kaspasy metabolismus MeSH
- kontrolní body buněčného cyklu účinky léků MeSH
- kyseliny karboxylové chemie MeSH
- lidé MeSH
- membránový potenciál mitochondrií účinky léků MeSH
- mikrotubuly účinky léků metabolismus MeSH
- nádorové buněčné linie MeSH
- proliferace buněk účinky léků MeSH
- signální transdukce účinky léků MeSH
- vztah mezi dávkou a účinkem léčiva MeSH
- zlomy DNA účinky léků MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
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 μM 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 μM 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.
- MeSH
- alkaloidy chemie farmakologie terapeutické užití MeSH
- Amaryllidaceae chemie metabolismus MeSH
- antitumorózní látky fytogenní chemie farmakologie terapeutické užití MeSH
- apoptóza účinky léků MeSH
- Ehrlichův tumor farmakoterapie mortalita patologie MeSH
- Kaplanův-Meierův odhad MeSH
- léky antitumorózní - screeningové testy MeSH
- lidé MeSH
- myši MeSH
- nádorové buněčné linie MeSH
- proliferace buněk účinky léků MeSH
- transplantace heterologní MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
