6 research outputs found
The cytotoxic and antitumoral effects of Remdesivir, an antiviral RdRp inhibitor, on different cancer cells in vitro
No full textBackgroundPrevious studies have shown similarities in the metabolism of cancer cells and parasites, suggesting that antiparasitic drugs may be used as anticancer agents. Remdesivir (Rem), an RdRp inhibitor, has been recently used in SARS-CoV-2 pandemic. Although the apoptotic effect of Rem has been demonstrated on the SKOV3 ovarian cancer cell line, its cytotoxic effect has not been analyzed in different cancer cells.ObjectiveWe aimed to evaluate its cell death-inducing effects on PC3 prostate cancer, HepG2 hepatocellular carcinoma, and A2058 malignant melanoma cells for the first time, using WST-1, Annexin V, cell cycle analysis, AO/EB staining, live-cell imaging, TEM, and gene expression analysis.ResultsRem treatment at 10, 25, and 50 & mu;M significantly decreased cell viability in all cancer cells (p < 0.01). While Rem triggered apoptosis in PC3, vacuole-dependent cell death was detected in HepG2 cells by visualizing the cells with TEM and time-dependent live-cell imaging. Moreover, both forms of cell death were triggered together in A2058. The formation of AVOs were observed more after 12 h and 24 h in HepG2 and A2058 cells, respectively. Additionally, Rem significantly induced cell cycle arrest in the G2/M (p < 0.01). Finally, the mRNA levels of autophagic markers, Atg12, Atg5, p62, Beclin, and LC3, were significantly increased in A2058 and HepG2 (p < 0.01) compared to control groups.ConclusionsOur results suggest that Rem has promising cytotoxic effects on various cancer cells. However, it triggers different types of cell death in different cancer types, indicating that further studies should focus on clarifying the molecular mechanism of the specific action of Rem
Synergistic anti-tumorigenic effects of cabazitaxel and usnic acid combination on metastatic castration-resistant prostate cancer cells
No full textBackground Prostate cancer (PC) affects millions of men, causing high mortality rates. Despite the treatment approaches, the options for metastatic castration-resistant prostate cancer (mCRPC), a lethal form of advanced PC, are still limited. Cabazitaxel (Cbx) is the last taxane-derived chemotherapeutic approved for Docetaxel-resistant mCRPC patients. However, its effects are limited due to the activation of several pathways. Therefore, new approaches are needed to increase the efficacy of Cbx. Usnic acid (UA) is a natural product with well-known anti-tumorigenic and synergistic effects with various chemotherapeutics. Although the cytotoxicity of UA and Cbx has been evaluated on mCRPC cells, the anti-tumorigenic effect of UA combination with any taxane has not been investigated yet. Thus, we aimed to evaluate the possible synergistic effect of Cbx+UA in mCRPC cells. Methods Cell viability and apoptosis were analyzed using WST-1 and Annexin-V. Morphological changes were visualized by fluorescent staining. Finally, cell cycle, mitochondrial health, and ROS levels were determined. Results Based on WST-1 results, 25 mu M UA exhibited significant additive and synergistic effects with the use of Cbx. Annexin V and cell cycle results showed that UA significantly enhanced the Cbx efficacy at increasing doses compared to using only Cbx (**p<0.01). Moreover, combined treatment significantly increased ROS levels and mitochondrial membrane depolarization compared with Cbx alone (**p<0.01). Conclusions Thus, the results suggest that UA increased the anti-tumorigenic effects of Cbx on mCRPC cells by increasing apoptosis, causing an increase in intracellular ROS and disrupting mitochondrial health. Consequently, combining UA and Cbx offers a new combined therapeutic strategy for mCRPC treatment
[MSB-10] The biomarker potential of serum-derived exosomes carrying autophagic regulators in carotid artery stenosis patients
No full textThis study aimed to investigate the biomarker potential of exosomes as autophagic regulators for discriminating ulcerated from smooth ones by serum-derived exosome isolation and real-time quantitative polymerase chain reaction (RT-qPCR) analysis.Eight carotid artery stenosis (CAS) patients (6 males, 2 females; mean age: 65.7±11 years) were included in the study conducted. Plaque ulceration was determined by Doppler ultrasonography. Afterward, the serum was separated and exosome/exosomal RNA isolation was performed using an exoRNeasy Midi Kit. Transmission electron microscopy and nanoparticle tracking analysis were performed for exosome characterization. Finally, the expression of autophagy-related genes (Atg5, Atg12, Beclin, and LC3) was analyzed by RT-qPCR.Five (62.5%) patients had ulcerated carotid plaques. The patients' stenosis degree was 70% or greater. Based on the RT-qPCR (120 nm), Atg12, Beclin, LC3, and Atg5 levels were downregulated by 3.79-, 5.28-, 2.40- (p>0.05), and 4.83-fold (p=0.02) in ulcerated plaques, respectively.Although further validation is needed, data suggests that autophagy-related mRNA cargoes of serum-derived exosomes may be potential biomarkers for risk stratification based on plaque surface morphology in CAS patients
Second-generation BRAF inhibitor Encorafenib resistance is regulated by NCOA4-mediated iron trafficking in the drug-resistant malignant melanoma cells
No full textAbstract The current study established the first in vitro Encorafenib resistance protocol in BRAF-mutated malignant melanoma (MM) cells and investigated the resistance-related mechanisms. After establishing Encorafenib-resistant A375-MM cells, resistant-related mechanisms were investigated using WST-1, Annexin V, cell cycle, morphological analysis, live-cell, Western blot, RNA-Seq, transmission electron microscopy-(TEM), oxidative stress and iron colorimetric assay. The most resistant group, called A375-R, was determined in the cells treated with a constant dose of 10 nM over 3 months. The viability, apoptosis, and G0/G1 arrest reflected the acquired chemoresistance. Autophagic Beclin and LC3 proteins, and AKT signaling increased in the A375-R. RNA-Seq results also exhibited altered epigenetic regulation of resistance; particularly ferritin family members, ion transport pathways. Then, increased NCOA4, FTH1, and iron levels detected in A375-R suggest that the iron metabolism-related mechanism, such as ferritinophagy, might be triggered, which was supported by TEM and oxidative stress analysis. Iron storage, transport, and ferritinophagy have the promising potential to be targeted for combining with BRAF-targeted therapy to reverse Encorafenib resistance in MM. Moreover, this is the first study evaluating in vitro Encorafenib resistance mechanisms, and we suggest that our findings contribute to improving new drug combinations targeting BRAF and iron metabolism in different MM cells
Ülsere plaklı yüksek riskli asemptomatik karotis arter stenozu hastalarının ayırımı için biyobelirteç profillemesi: Pilot çalışma
No full textAlthough various methods are used to treat patients with asymptomatic carotid artery stenosis (ACAS), approaches are controversial, and combining imaging of carotid plaque features with biomarkers to identify plaques prone to rupture may be crucial in identifying high-risk ACAS patients. This study aimed to investigate a blood-based biomarker for discriminating ulceration in ACAS patients by analyzing plaque surface morphology through RNA sequencing of blood samples. Peripheral blood samples were collected from ACAS patients with plaque morphology determined by Doppler ultrasonography. Then, total RNA was isolated, and RNA-Seq was performed to analyze differentially expressed genes (DEGs). The KEGG, Reactome, and Gene Ontology (GO) terms pathway enrichment analyses were performed to investigate the molecular functions and biological processes involved in plaque formation. The pilot study included 7 ACAS patients, 57.1 % exhibiting ulcerated plaques. RNA-Seq results revealed significant upregulation of genes related to immune response, cell cycle regulation, and oxidative stress in ulcerated plaques. Especially, TP73, CCL3L3, and PXDNL genes showed the highest fold changes, indicating their role in endothelial damage, immune activation, and oxidative stress. KEGG and Reactome analyses identified TNF and chemokine signaling pathways as key regulators in ulcerated plaque formation. Our findings indicate that TP73, CCL3L3, and PXDNL may be potential biomarkers for identifying high-risk ACAS patients with ulcerated plaques due to their involvement in immune system regulation and oxidative stress-related processes. Thus, these genes and the pathways may be candidate biomarkers for early diagnosis and risk stratification, improving treatment approaches for ACAS.Asemptomatik karotis arter stenozu (ACAS) olan hastaların tedavisinde çeşitli yöntemler kullanılsa da yaklaşımlar tartışmalıdır ve plak rüptürü eğilimli olan hastaları belirlemek için karotis plak görüntülerinin biyobelirteçlerle birleştirilmesinin, yüksek riskli ACAS hastalarının belirlenmesinde önemli olduğu düşünülmektedir. Mevcut çalışmada, plak yüzey morfolojisini analiz ederek ACAS hastalarında ülserasyonu ayırt etmek için RNA sekanslama yoluyla kan bazlı biyobelirteç araştırılması amaçlanmıştır. Bu doğrultuda, Doppler ultrasonografi ile plak morfolojisi belirlenen ACAS hastalarından periferik kan örnekleri toplandı. Daha sonra, total RNA izole edildi ve farklı şekilde ifade edilen genleri (DEG'ler) analiz etmek için RNA-Seq gerçekleştirildi. Plak oluşumunda yer alan moleküler işlevleri ve biyolojik süreçleri araştırmak için KEGG, Reactome ve Gen Ontolojisi (GO) ile yolak zenginleştirme analizleri gerçekleştirildi. Pilot çalışmaya 7 ACAS hastası dahil edildi ve %57,1'inde ülsere plaklar olduğu görüldü. RNA-Seq sonuçları ülsere plaklarda bağışıklık tepkisi, hücre döngüsü düzenlemesi ve oksidatif stresle ilgili genlerde önemli bir artış olduğunu ortaya koydu. Özellikle TP73, CCL3L3 ve PXDNL genlerinin bu süreçlerde rol aldığı ve bu genlerin endotel hasarı, bağışıklık aktivasyonu ve oksidatif streste rolleri olduğu gösterildi. KEGG ve Reactome analizleri, ülserli plak oluşumunda anahtar düzenleyiciler olarak TNF ve kemokin sinyal yollarını tanımladı. Bulgularımız, TP73, CCL3L3 ve PXDNL'nin bağışıklık sistemi düzenlemesi ve oksidatif stresle ilişkili süreçlerdeki katılımları nedeniyle ülserli plakları olan yüksek riskli ACAS hastalarını tanımlamak için potansiyel biyobelirteçler olabileceğini göstermektedir. Bu nedenle, ilişkili genlerin ve yolakların erken tanı ve risk sınıflandırması için aday biyobelirteçler olabileceği ve ACAS için tedavi yaklaşımlarını iyileştirebileceğini düşündürmektedir
Effect of pain-related sodium channels <i>scn9a</i> and <i>scn10a</i> polymorphisms in migraine chronification
No full textObjectiveBased on the frequency of attacks, migraine is classified into two subtypes: episodic (EM) and chronic (CM). Migraine progression from EM to CM is supposed to be affected by various factors, which is known as "migraine chronification. However, the pathophysiology of migraine chronification is multifactorial and still not fully understood. Ion channels are hypothesized to play a role in migraine pathophysiology and are essential for generating and suppressing action potentials, which lie under the pain and related symptoms. Two sodium channel genes, SCN9A and SCN10A, have been reported to be associated with various pain sensitivities. Studies have shown that patients with EM and CM are more sensitive to chronic pain and that pain sensitivity may be a risk factor for chronification. Thus, the current study aimed to determine whether pain sensitivity-related SCN9A and SCN10A polymorphisms affect the EM-to-CM transition.MethodsFor this purpose, genotyping was performed using TaqMan SNP Assay for the (i) SCN9A rs7595255, rs12622743, and rs11898284, (ii) SCN10A rs6795970, rs6801957 SNPs in Turkish EM and CM patients.ResultsThe results showed that SCN9A and SCN10A polymorphisms did not play a role in the development of chronicity. However, the results indicated that the SCN10A rs6795970 polymorphism was associated with osmophobia (p = 0.036).ConclusionSCN10A rs6795970 polymorphism may be necessary to predict the EM-to-CM transition and identify therapeutic targets. However, further studies are required to confirm the osmophobia association of the SCN10A rs6795970 polymorphism and to investigate the role of these SNPs in chronification
