Studier & Utbildning

Oscar mejlade över en ny studie om trippelnegativ bröstcancer. En riktigt elak bröstcancer med dåliga odds för överlevnad.

Epidermal growth factor potentiates EGFR(Y992/1173)-mediated therapeutic response of triple negative breast cancer cells to cold atmospheric plasma-activated medium

Abstract

Cold atmospheric plasma (CAP) holds promise as a cancer-specific treatment that selectively kills various types of malignant cells. We used CAP-activated media (PAM) to utilize a range of the generated short- and long-lived reactive species. Specific antibodies, small molecule inhibitors and CRISPR/Cas9 gene-editing approaches showed an essential role for receptor tyrosine kinases, especially epidermal growth factor (EGF) receptor, in mediating triple negative breast cancer (TNBC) cell responses to PAM. EGF also dramatically enhanced the sensitivity and specificity of PAM against TNBC cells. Site-specific phospho-EGFR analysis, signal transduction inhibitors and reconstitution of EGFR-depleted cells with EGFR-mutants confirmed the role of phospho-tyrosines 992/1173 and phospholipase C gamma signaling in up-regulating levels of reactive oxygen species above the apoptotic threshold. EGF-triggered EGFR activation enhanced the sensitivity and selectivity of PAM effects on TNBC cells. The proposed approach based on the synergy of CAP and EGFR-targeted therapy may provide new opportunities to improve the clinical management of TNBC.

Jag går direkt till forskarnas användande av HoloMonitor.

 Representative HoloMonitor images (taken every 5 min, 3 days in total) of MDA-MB-231 cell examples treated with EGF, 50 % 10PAM, or EGF+50 % 10PAM. The time points of cell division (Control and EGF group) and apoptosis (PAM and PAM+EGF group) have been selected for display. d Cell area (μm2) and optical thickness avg (μm) of MDA-MB-468 and MDA-MB-231 cells measured by the Holomonitor Hstudio after EGF, PAM or EGF+PAM treatments as in (a) above, for 3 days, versus Control in MDA-MB-468 and MDA-MB-231 cultures.

Morphology of MDA-MB-231 cells as viewed in the Holomonitor changed from a spindle-like shape to a small and roundish shape after the combined treatment of 50 % 10PAM and EGF for 3 h (Fig. 3c). While MDA-MB-231 cell division in the control group was observed at 12 h, PAM-induced cell death was evident at 8 h and cell death was initiated at 3 h when cells were exposed to combined EGF and 50 % 10PAM treatment. Under the control conditions, the correlation between cell area (μm2) and optical thickness avg (μm) as measured by Holomonitor Hstudio was evident in video imaging (Fig. 3d and Fig. S3a-b, S4a-b). 

Among cells receiving combined PAM and EGF exposure, 74 % (32 % + 42 %) underwent apoptosis and only 2 % underwent necrosis (p < 0.0001), which was consistent with Holomonitor observations.

HoloMonitor imager measurements: SUM159 and MDA-MB-231 cells were plated in 96-well plates at 5000 cells per well for 24 h before real-time monitoring. When the cell density was around 30 %, the digital holograms of cells were set up using the HoloMonitor M4 Digital Holography Cytometer (Phase Holographic Imaging PHI AB, Lund, Sweden). The results were calculated using Hstudio M4 software (Phase Holographic Imaging PHI AB, Lund, Sweden). 

Scrolla ner till slutet i studien.Där hittar ni 7 olika sekvenser med cellrörelser på dessa uschliga cancerceller med och utan cancerläkemedel.

-------------------------------------------------------------------------------------------------------------

I en annan aktuell studie har koreanska forskare studerat huruvida ämnen från vissa svampar kan få en bröstcancer att stanna upp i sin utveckling och tillväxt. Dvs tillföra ett ämne från svampar och få stopp på cancercellernas migrering. Ett annorlunda grepp som absolut är intressant.Kan man kalla det old school medicine? Fjärran Österns traditionsrika kunskaper och mångårigt användande av medicinalväxter ska aldrig förringas.

Induction of Fungal Secondary Metabolites by Co-Culture with Actinomycete Producing HDAC Inhibitor Trichostatins

 Published November 28, 2023

A recently bioinformatic analysis of genomic sequences of fungi indicated that fungi are able to produce more secondary metabolites than expected. Despite their potency, many biosynthetic pathways are silent in the absence of specific culture conditions or chemical cues. To access cryptic metabolism, 108 fungal strains isolated from various sites were cultured with or without Streptomyces sp. 13F051 which mainly produces trichostatin analogues, followed by comparison of metabolic profiles using LC-MS. Among the 108 fungal strains, 14 produced secondary metabolites that were not recognized or were scarcely produced in mono-cultivation. Of these two fungal strains, Myrmecridium schulzeri 15F098 and Scleroconidioma sphagnicola 15S058 produced four new compounds (1-4) along with a known compound (5), demonstrating that all four compounds were produced by physical interaction with Streptomyces sp. 13F051. Bioactivity evaluation indicated that compounds 3-5 impede migration of MDA-MB-231 breast cancer cells.

Introduction 

Fungi are major composers of ecosystems and producer of new secondary metabolites that are used in medicine and agriculture. To date, 100,000 fungi have been identified and approximately 5 million fungal species may exist. A recent whole-genome sequencing analysis indicated that fungi have the potential to produce more secondary metabolites than expected. Despite this genetic capability, many biosynthetic pathways are silent or the rediscovery of secondary metabolites is frequent under traditional laboratory culture conditions. Various strategies have been developed to access cryptic metabolites. Co-culture of two microorganisms in the same culture environment is a representative strategy. This strategy induces physical and metabolite interactions between the two microorganisms, which ultimately increases gene expression, and consequently increases metabolite production. Another approach is epigenetic manipulation at the chromatin level using histone deacetylase (HDAC) and DNA methyltransferase (DNMT) inhibitors, which regulate transcriptional gene expression in fungi. In a previous study, we examined the chemical profiles of actinomycetes isolated from soil samples collected from Ulleung Island by liquid chromatography-photodiode array-mass spectrometry (LC-PDA-MS). Among the 208 actinomycete strains isolated, Streptomyces sp. 13F051 produced trichostatin analogues. Trichostatin A (TSA) is a HDAC inhibitor, its mechanism of action has been established; the hydroxamate group of TSA chelates a zinc ion in the active site pocket of HDAC, blocking the catalytic reaction. Owing to its potent HDAC inhibitory activity, TSA has been used to enhance the cryptic secondary metabolites of fungi. We hypothesized that if fungi were cultured with an actinomycete, producing HDAC inhibitors, TSA analogues, novel fungal secondary metabolites are induced by HDAC inhibitors or physical interaction between two microorganisms. To test this hypothesis, various fungal strains were co-cultured with an actinomycete on agar plates, and their metabolic profiles were monitored by LC-MS. The co-culture of Streptomyces sp. 13F051 and the three fungal strains produced secondary metabolites that were not recognized or were scarcely produced in mono-cultures. To determine whether the induced metabolites were produced by HDAC inhibitors or physical interactions, the four fungal strains were cultured with a disk containing acetone extracts of Streptomyces sp. 13F051. The two fungal strains produced the novel secondary metabolites via physical interaction in the co-culture system. Here we report four new fungal secondary metabolites: dinapinone analogues (1 and 2), sambutoxin analogues (3 and 4) and a known compound AS2077715 (5) induced by physical interaction in the co-culture system. After isolation, all the compounds were further studied to elucidate their structure and biological activities. 

Materials and Methods

Time-lapse Cell Tracking Analysis. MDA-MB-231 cells (8 × 103 cells/100 l) were seeded on -Slide I (80106, ibidi, Germany). After 4 h, 900 l DMEM was added to the slide. After incubation for 16 h, cells were treated with indicated concentrations of compound and directly transferred to a HoloMonitor M4 time-lapse cytometer (Phase Holographic Imaging) kept in a 37°C incubator. Live cells were imaged every 15 min for 12 h. Migration was analyzed using the HoloStudio M4 software.

Fig. 6. Effects of compounds on cell mobility of MDA-MB-231 cells. (A) Cells were treated with 25 μM of each compound. Cell movement was captured every 15 min for 12 h. (B) Cell mobility analyzed with HoloStudio M4 software. 231 cells. After treatment with the indicated concentrations of the test compounds for 24 h, the rate of cells migrating to the empty area was significantly decreased by treatment with compounds 3, 4, and 5 in a dosedependent manner (Fig. 5). To further investigate the effect of the compounds on cell migration, we performed time-lapse cell tracking analysis. The cells were treated with 25 M of the indicated compounds. Cell movement was captured with a HoloMonitor M4 time-lapse cytometer every 15 min for 12 h. As expected, cell motility was markedly inhibited in cells treated with compounds 3 (44% inhibition), 4 (32% inhibition), and 5 (65% inhibition) compared to control cells treated with DMSO (Fig. 6).

--------------------------------------------------------------------------------------------------------

Nästa studie handlar om hudcancer.Metastaserande hudcancerceller som kan ställa till det utav h-e.
Forskarna har studerat hur ett visst ämne kan blocka dessa cancerceller från att växa till sig resulterande i metastaser. Ger cancerpatienten tid att få behandling och förhoppningsvis slippa dessa farliga dottertumörer.

IK Channel-Independent Effects of Clotrimazole and Senicapoc on Cancer Cells Viability and Migration

Abstract

Many studies highlighted the importance of the IK channel for the proliferation and the migration of different types of cancer cells, showing how IK blockers could slow down cancer growth. Based on these data, we wanted to characterize the effects of IK blockers on melanoma metastatic cells and to understand if such effects were exclusively IK-dependent. For this purpose, we employed two different blockers, namely clotrimazole and senicapoc, and two cell lines: metastatic melanoma WM266-4 and pancreatic cancer Panc-1, which is reported to have little or no IK expression. Clotrimazole and senicapoc induced a decrease in viability and the migration of both WM266-4 and Panc-1 cells irrespective of IK expression levels. Patch-clamp experiments on WM266-4 cells revealed Ca2+-dependent, IK-like, clotrimazole- and senicapoc-sensitive currents, which could not be detected in Panc-1 cells. Neither clotrimazole nor senicapoc altered the intracellular Ca2+ concentration. These results suggest that the effects of IK blockers on cancer cells are not strictly dependent on a robust presence of the channel in the plasma membrane, but they might be due to off-target effects on other cellular targets or to the blockade of IK channels localized in intracellular organelles.

Även här går jag direkt till forskarnas användande av HoloMonitor.Bilden visar cancerceller med och utan cancerläkemedel.

4.6. Scratch-Healing Assays

Panc-1 and WM266-4 cells were seeded in 35 mm petri dishes (2.5 × 105 cells/petri). Then, 24 h after seeding, cells reached 100% confluency, and a wound in the cells layer was created with a 200 µL pipette tip. The cells’ medium was then removed and substituted with new medium enriched with 30 µM clotrimazole, 30 µM senicapoc, or the corresponding volume of DMSO. Cell migration was monitored in time under a holographic microscope Holomonitor M4 live cell imaging system (Phase Holographic Imaging, PHI AB, Lund, Sweden). Data analysis was performed using HStudio (PHI AB, Lund, Sweden). Pictures were taken after 24 h. Data are reported as increase in cell-covered areas after 24 h with respect to the control condition (DMSO).

Cell migration from the scratch edges was monitored with a holographic microscope (see Supplementary Figure S1B). Figure 2B shows the movement of scratch edges as increases of cell-covered areas after 24 h long treatment with 30 µM clotrimazole or 30 µM senicapoc with respect to control DMSO-treated cells. In both cell lines, we observed a high variability in wound edges after 24 h long exposure to 30 µM clotrimazole. From experiments with senicapoc, we obtained more uniform datasets which showed that wound healing was significantly lower than that of DMSO-treated cells for both WM266-4 and Panc-1. Similarly to trans-well migration, WM226-4 cells were more affected by senicapoc than Panc-1 cells, which were nevertheless sensitive to the molecule. This indicated that the reduction in WM266-4 cell migration is possibly caused by a combination of IK blockage and an IK-independent effect, the latter being responsible for the Panc-1 migration decrease.

-----------------------------------------------------------------------------------------------------------

Sen har kinesiska forskare gått vidare med sina studier om hur få stopp på den överjävliga cancern glioblastoma. Forskarna utgår från tesen att strypa näringstillförsel (blod) till cancercellerna, alltså en utveckling av de tidigare studierna bloggen berättat om.

m5C modification of LINC00324 promotes angiogenesis in glioma through CBX3/VEGFR2 pathway

Abstract

Angiogenesis plays a major role in tumor initiation, progression, and metastasis. This is why finding antiangiogenic targets is essential in the treatment of gliomas. In this study, NSUN2 and LINC00324 were significantly upregulated in conditionally cultured glioblastoma endothelial cells (GECs). Knockdown of NSUN2 or LINC00324 inhibits GECs angiogenesis. NSUN2 increased the stability of LINC00324 by m5C modification and upregulated LINC00324 expression. LINC00324 competes with the 3’UTR of CBX3 mRNA to bind to AUH protein, reducing the degradation of CBX3 mRNA. In addition, CBX3 directly binds to the promoter region of VEGFR2, enhances VEGFR2 transcription, and promotes GECs angiogenesis. These findings demonstrated NSUN2/LINC00324/CBX3 axis plays a crucial role in regulating glioma angiogenesis, which provides new strategies for glioma therapy.

Tyvärr har sajten låst in nästan hela studien nu så partierna där forskarna berättar om användande av HoloMonitor kommer man inte åt i dagsläget. Undertecknad skulle varit snabb med att kopiera den infon,men men...

-----------------------------------------------------------------------------------------------------

Avslutningsvis kan berättas att Mr hardworking Goran Dubravcic satt personalen på Warszawa baserade försäljningsstället Perlan i Polen på HoloMonitor-utbildning. 

Perlans hemsida inleds med :

Gör kopplingen till bjässen Agilent nästan övertydlig. 😎

                                             Mvh the99

Som service till alla ev HoloMonitornyfikna forskare : HoloMonitor Demo