Forskningsrapport från Taiwan

En studie från Taiwan kommer snart publiceras.Den väntar på granskning innan den publiceras i ett vetenskapligt fora.10 forskare från Taiwan (9st) och Kina (1st) står bakom en mkt intressant cancerstudie.

Forskarna från Taiwan utgår från det HoloMonitor-utrustade universitet National Cheng Kung University.

Sagda universitet ståtar med minst 2 HoloMonitor,det senaste inköpt för 1 år sen.Bloggen uppmärksammade universitet redan 2018. Läs det inlägget för intressant bakgrund och nuläge.

I nu aktuell studie berättar de om helt nya rön beträffande ett protein som ingår i människas genuppsättning.

Proteinet ingår i genen Wwox och funktionen är sen tidigare klarlagd. "Loss of WWOX gene leads to severe neural diseases and metabolic disorders in newborns." Forskarna har nu upptäckt att samma gen har påverkan på en cancercells utveckling.Sen tidigare har forskare visat vilken inverkan den har på cancertumörer,att det går att minska deras utbredning i kropp genom att påverka/manipulera genen.

Men i denna studie har de visat på att det går att påverka en enskilds cancercells utveckling via (annan) påverkan/manipulering av genen Wwox. Genom idogt användande av HoloMonitor har de observerat  cancerceller och den utveckling de genomgår när genen påverkas.

Functional Wwox-Expressing Cells Respond to Uv/Cold Shock-Mediated Bubbling Cell Death, Whereas Dysfunctional Cells Undergo Explosion

Abstract

Majority of normal and benign cancer cells express functional WW domain-containing oxidoreductase (WWOX), designated WWOXf. Metastatic cancer cells develop WWOX deficiency or express dysfunctional WWOX (WWOXd). Loss of WWOX gene leads to severe neural diseases and metabolic disorders in newborns. Interestingly, UV irradiation or UV/cold shock induces non-apoptotic nuclear bubbling cell death (BCD) in WWOXf, and pop-out explosion death (POD) in WWOXd. When WWOXf cells were exposed to UV and then incubated at room temperature, these cells rapidly had major shutdown of mRNA and protein synthesis machinery in less than one minute. Thirty minutes later, formation of a nitric oxide (NO)-containing nuclear bubble per cell occurred, followed by eventual cell death without caspase activation and chromosomal DNA fragmentation. UV signaled the co-translocation of WWOX and TRAF2 complex to the nucleus, where the prosurvival TRAF2 blocked the proapoptotic WWOX via its zinc finger region. By time-lapse holography, cell thickness was increased in dividing and apoptotic WWOXf, whereas the cell thickness was not increased during BCD in WWOXf. Gene chip analysis for mRNA profiling showed that glutathione peroxidase GPX1/2 and RNA-regulated  protein  kinase PKR were involved in BCD in L929 fibroblasts, a WWOXf cell line. In parallel, upon UV exposure, both GPX1/2 and PKR proteins were significantly downregulated in skin and liver of hairless mice, suggesting that UV-induced WWOX/TRAF2 signaling limits the expression of GPX1/2 and PKR for leading to cell and organ damage and death.

En googleöversättning ger lite mer dramatik till innehållet.

Majoriteten av normala och benigna cancerceller uttrycker funktionellt WW-domäninnehållande oxidoreduktas (WWOX), betecknat WWOXf. Metastaserande cancerceller utvecklar WWOX-brist eller uttrycker dysfunktionell WWOX (WWOXd). Förlust av WWOX-genen leder till allvarliga neurala sjukdomar och metabola störningar hos nyfödda. Intressant nog inducerar UV-bestrålning eller UV/kylchock icke-apoptotisk nukleär bubblande celldöd (BCD) i WWOXf, och pop-out explosion death (POD) i WWOXd. När WWOXf-celler exponerades för UV och sedan inkuberades vid rumstemperatur, hade dessa celler snabbt stor avstängning av mRNA och proteinsyntesmaskineri på mindre än en minut. Trettio minuter senare inträffade bildning av en kväveoxid (NO)-innehållande kärnbubbla per cell, följt av eventuell celldöd utan kaspasaktivering och kromosomal DNA-fragmentering. UV signalerade samtranslokationen av WWOX och TRAF2-komplex till kärnan, där den prosurvival TRAF2 blockerade den proapoptotiska WWOXen via sin zinkfingerregion. Genom time-lapse holografi ökades celltjockleken i delande och apoptotisk WWOXf, medan celltjockleken inte ökades under BCD i WWOXf. Genchipanalys för mRNA-profilering visade att glutationperoxidas GPX1/2 och RNA-reglerat proteinkinas PKR var involverade i BCD i L929-fibroblaster, en WWOXf-cellinje. Parallellt, vid UV-exponering, var både GPX1/2- och PKR-proteiner signifikant nedreglerade i hud och lever hos hårlösa möss, vilket tyder på att UV-inducerad WWOX/TRAF2-signalering begränsar uttrycket av GPX1/2 och PKR för att leda till cell- och organskador och döden.

 

Från den medföjande PDF-filen fås mer info.

 

1. Introduction 

Although it was originally recognized as a candidate tumor suppressor (Chang, et al., 2007), WW domain-containing oxidoreductase (WWOX) plays an essential role in controlling the development of neural system and preventing neurodegeneration (Chen et al., 2004; Sze et al., 2004; Chang, et al., 2007; Chang and Chang, 2015; Cheng et al., 2020). WWOX limits the progression of Alzheimer’s disease and other neurodegenerative diseases. Functional deficiency of WWOX gene in newborns leads to severe neural disease, seizures, metabolic disorder and early death (Aldaz et al., 2020; Cheng et al., 2020; Repudi et al., 2021). WWOX gene has recently been regarded as one of the risk factors for Alzheimer’s disease (Kunkle et al., 2019). When WWOX is Y33 phosphorylated, pY33-WWOX supports normal physiology, inhibits neurodegeneration, and blocks cancer growth. When pY33-WWOX is downregulated and pS14- WWOX upregulated, pS14-WWOX supports cancer growth and promotes the progression of Alzheimer’s disease (Huang and Chang, 2018). Thus, WWOX exhibits two faces in managing the progression of Alzheimer’s disease, cancer growth, and probably other diseases.

 

In contrast to the caspase-dependent or -independent programmed cell death (Fink and Cookson, 2005), we have previously reported the nucleus-initiated cell death, designated bubbling cell death (BCD) (Chen et al., 2015; Chang, 2016). When temperature is lowered in the surrounding environment, dying cells do not undergo apoptosis and yet they undergo BCD (Chen et al., 2015; Chang, 2016). Our study concerns the severity of frostbite and UV irradiation in the polar regions and even outer space (Chen et al., 2015; Chang, 2016). When exposed to severe cold and strong UV irradiation, people suffer serious damages to their skin and lead to further and rapid damage in internal organs, and eventually result in limb amputations, organ failure, and even death. BCD has been defined as “formation of a single bubble from the nucleus per cell and release of this swelling bubble from the cell surface to extracellular space that irreversibly causes cell death” (Chen et al., 2015; Chang, 2016). When cells are subjected to UV irradiation and/or brief cold shock (4°C for 5 min) and then incubated at room temperature or 4°C for time-lapse microscopy, each cell releases an enlarging nuclear gas bubble containing nitric oxide (NO). Certain cells may simultaneously eject hundreds or thousands of extracellular vesicles (EV) or exosome-like particles.  

Unlike apoptosis, BCD does not exhibit membrane phosphatidylserine flip-over, mitochondrial apoptosis, damage to Golgi complex, and chromosomal DNA fragmentation.When the temperature is increased back at 37°C, BCD stops and apoptosis starts (Chen et al., 2015; Chang, 2016). Indeed, BCD can occur at 37°C. For example, when cells are transiently overexpressed with the Hyal-2/WWOX/Smad4 signaling complex, high-molecular-weight hyaluronan induces BCD at 37°C (Hsu et al., 2017).

 

Time-lapse holography forskarna relaterar till i studien är alltså från HoloMonitor.

2.3. Time-lapse bright field and fluorescent microscopy and time-lapse holography (verifiering)

Further, UV/cold shock-treated cells were imaged by time-lapse holographic microscopy (HoloMonitor M4) and analyzed by Hstudio M4 Tracking software. 

 

3.5. Dramatic increase in cellular thickness occurs during apoptosis but not BCD, as determined by timelapse holographic microscopy 

Next, we investigated cell morphological changes during BCD mainly by time-lapse holographic microscopy.  

 

4.4. Normal cells tall up during division and apoptosis 

By time-lapse holography, we have demonstrated for the first time that there is an increased thickness or height in dividing or apoptotic cells. Cell volume increase is normal in dividing cells. Nonetheless, the rationale for cells to get taller and then divide remains to be established. The tallness lasts less than 20 min and then the cell divides. In stark contrast, during apoptosis, the increased cell height can last 5 to 8 hr, followed by breakdown of the whole cell. However, during BCD, the bubble released from the nucleus is indeed flattened, compared to the cell body, suggesting that the nuclear bubble contains not only NO but also cytoplasmic liquid. These observations were done with WWOXf L929S and COS7 cells. Whether WWOXd cells tall up prior to explosion remains to be established.

Fig. 2. Time-lapse holographic microscopy shows dramatically increases cell thickness during apoptosis, but not BCD. (A,B) COS7 cells were exposed to UV and then cold shock at 4oC for 12 hr. Cell bubbles were observed, as determined by light and holographic microscopy, respectively. 

 

Results 

WWOXf cells undergo nuclear bubbling during BCD at room temperature and below We have identified a panel of WWOXf cells, which are TNF-sensitive L929S fibrosarcoma cells, human breast MCF7 cancer cells, monkey kidney COS7 fibroblasts, human prostate DU145 cancer cells, human colon HCT116 cells, human squamous cell carcinoma SCC9 and SCC15 cells, mouse embryonic fibroblasts (MEF), and other cancer cells or any human and mouse normal cells (Chen et al., 2021). Four representative WWOXf cell lines, including L929S, DU145, HCT116, MCF7, and Wwox+/+ wild type MEF, were exposed to UV irradiation (960 mJoule/cm2 ) and then subjected to time-lapse microscopy at room temperature.  

Min kommentar

"Värdet" av denna studie kan jag som lekman naturligtvis inte ha nån åsikt om. Men att forskarna lyckats nå nya insikter i cancereländet samt deras idoga användande av HoloMonitor för att nå dit applåderar jag såklart.

 

Bra jobbat - Good job - 好作品  säger jag till de 10 forskarna.

                                       Mvh the99

 

Som service till alla ev HoloMonitornyfikna forskare : PHIAB Webshop