måndag 14 juni 2021

Ny forskningsrapport från Kina

8 kinesiska forskare har fått sina studier om en cancer med dåliga överlevnadschanser granskade,godkända och publicerade.Det handlar om levercancer (hepatocellular carcinoma) som skördar 800 000 människoliv om året.Denna cancertyp är riktigt lömsk då den oftast inte upptäcks förrän det är försent att sätta in räddande behandling.Man opererar om så går och tar då bort tumören eller i värsta fall hela levern.Det brukar resultera i behov av en levertransplantation.Har cancern gått för långt kan man inte operera utan ger patienten läkemedel som dessväre inte visat sig ha nån större effekt.
Överlevnad vid denna behandlingsform sträcker sig ca 3 månader från diagnos.

Behovet att ett helt nytt läkemedel är mao stort och det är med den målsättningen dessa forskare har bedrivit sina studier.Intressant är att de gått tillbaka till kinesisk urminnesmedicin,dvs extract från örter osv..Här har de använt sig av en substans från roten ginseng,20(S)-protopanaxadiol (20(S)-PPD) och fått fram lovande resultat.
Forskarna har behövt använda sig av PHI`s HoloMonitor för att kunna studera cancerceller behandlade med substansen.Det inne i en inkubator under 24 timmar med 30 minuters intervaller.

Received 1 April 2021, Revised 25 May 2021, Accepted 1 June 2021, Available online 10 June 2021.

Abstract

Background

20(S)-protopanaxadiol (20(S)-PPD), one of the main active metabolites of ginseng, performs a broad spectrum of anti-tumor effects. Our aims are to search out new strategies to enhance anti-tumor effects of natural products, including 20(S)-PPD. In recent years, fasting has been shown to be multi-functional on tumor progression. Here, the effects of fasting combined with 20(S)-PPD on hepatocellular carcinoma growth, apoptosis, migration, invasion and cell cycle were explored.

Methods

CCK-8 assay, trypan blue dye exclusion test, imagings photographed by HoloMonitorTM M4, transwell assay and flow cytometry assay were performed for functional analyses on cell proliferation, morphology, migration, invasion, apoptosis, necrosis and cell cycle. The expressions of genes on protein levels were tested by western blot. Tumor-bearing mice were used to evaluate the effects of intermittent fasting combined with 20(S)-PPD.

Results

We firstly confirmed that fasting-mimicking increased the anti-proliferation effect of 20(S)-PPD in human HepG2 cells in vitro. In fasting-mimicking culturing medium, the apoptosis and necrosis induced by 20(S)-PPD increased and more cells were arrested at G0-G1 phase. Meanwhile, invasion and migration of cells were decreased by down-regulating the expressions of matrix metalloproteinase (MMP)-2 and MMP-9 in fasting-mimicking medium. Furthermore, the in vivo study confirmed that intermittent fasting enhanced the tumor growth inhibition of 20(S)-PPD in H22 tumor-bearing mice without obvious side effects.

Conclusion

Fasting significantly sensitized HCC cells to 20(S)-PPD in vivo and in vitro. These data indicated that dietary restriction can be one of the potential strategies of chinese medicine or its active metabolites against hepatocellular carcinoma.



Materials and Methods (urval)


2.7. HoloMonitorTM M4 analysis

The HoloMonitorTM M4 (Phase Holographic Imaging, Lund, Sweden), placed in a standard cell culture incubator, was used to record proliferation, track and related parameters of HepG2 cells which were exposured to 20(S)-PPD in control medium or fasting-mimicking medium in 6-well plates. Imaging was conducted every 30 mins for 24 h and analyzed by HoloMonitor software.



(D) The cell morphology analyzed by HoloMonitorTM M4 and real-time holographic imaging were recorded at the doses indicated (24 h). Differences were considered as significant when P < 0.05 (∗) or P < 0.01 (∗∗) or P < 0.001 (∗∗∗).


3D reconstruction imaging (Fig. 4) has an advantage for the comparison of cell volumn and morphology (i.e., thickness). Acquisition of mesenchymal phenotype through EMT has been conformed to be associated with aggressive tumor subtypes and poor clinical outcome in patients. It was observed that cells in fasting-mimicking medium tended to inhibit HepG2 cell mesenchymal phenotype. Next, we tracked the movement of HepG2 cells in control medium or fasting-mimicking medium combined with 20(S)-PPD by the HoloMonitor M4 time-lapse cytometer. Time-lapse cell-tracking analysis confirmed that fasted cells showed lower random motility and speed with 20 μM 20(S)-PPD.
 

 
 
Fig. 4. The motility was anlyzed by real-time holographic imaging.(A) The range of cell movement was analyzed. And the area and volumn of cells were also analyzed by HoloMonitorTM M4(B–C). (D) Motility speed of cells were analyzed. Differences were considered as significant when P < 0.05 (∗) or P < 0.01 (∗∗) or P < 0.001 (∗∗∗).

4. Discussion

Over the last two decades only 14.1% in 5-year survival rate have been achieved for patients with HCC, which represents one of the most aggressive tumors due to its rapid progress. At present, cytotoxic chemotherapy remains the major treatment option for advanced HCC. Some patients are allowed to accept targeted therapy, such as sorafinib. However, the objective response rate (ORR) of sorafenib for advanced HCC is 5% at most. Moreover, sorafenib still shows adverse effects such as poor tumor-shrinking effects and relatively high toxicity (e.g., hand-foot skin reaction).

20(S)-protopanaxadiol (20(S)-PPD), a derivative of ginsenosides, is the final form of protopanaxadiol saponins metabolized by human intestinal microflora. 20(S)-PPD performs many biological functions, including differentiation regulation of neural stem cells and anti-inflammatory properties

The major novel findings in the present study were that 20(S)-PPD performed a role in a variety of tumor inhibition. But here's the question, the effective concentration of 20(S)-PPD is relatively high. For example, It was reported that 20(S)-PPD inhibited the viability of HepG2 cell in a dose- and time-dependent manner. But the IC50 values were up to 81.35, 73.5, 48.79 μM at 24, 48 and 72 h, respectively. To enhance the bioavailability of 20(S)-PPD in tumors, novel, safe and effective methods are needed.

According to the cell morphology observed under a microscope, apoptosis, one of the biological processes results in cell death, is observed by a number of typical morphological changes in the structure of the cell (Fig. 2D). Additionally, data from flow cytometry confirmed that 20(S)-PPD induced more apoptosis and necrosis in fasting-mimicking medium. Importantly, fasting-mimicking-alone treatment did not show any significant effects on proliferation and death of HepG2 cells.
 
Based on the results of in vitro and in vivo experiments, our results provided an experimental foundation that fasting promoted anti-tumor effects of 20(S)-PPD, suggesting that restrictive dietary interventions might be a potential strategy for treatments against hepatocellular carcinoma. 
These extensively findings pave the way for future fasting studies in clinical treatment of tumors. 

Min kommentar
Vi skulle behöva en motsvarighet till GRETA som drev oponion att världens alla statsledare fick upp ögonen hur j-a (pardon my french) effektiv respektive lands cancerforskning skulle vara om deras cancerforskare fick tillgång till varsitt HoloMonitor instrument.Cancergåtan skulle lösas bra mkt snabbare är jag inte 99% säker på,utan 100% säker.
Ska vi alla phi,are försöka luska reda på en HANS i samma ålder som GRETA så det får max uppmärksamhet? 😎

                                                          Mvh the99
 
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