AKT1 Research Articles

Reversal of Mucin 1 Reduction-Induced Enterocyte Apoptosis by Retinoic Acid through the PI3K/AKT Signaling Pathway in an In vitro Model of Necrotizing Enterocolitis.

This study aimed to investigate the roles of Mucin 1 (MUC1), the PI3K/AKT pathway, and enterocyte apoptosis in Necrotizing Enterocolitis (NEC). Using an NEC Caco-2 cell model, retinoic acid treatment and MUC1 gene silencing were employed. Flow cytometry was used to assess apoptosis, while quantitative PCR and western blot analyses were conducted to evaluate the gene and protein expressions of MUC1, PI3K, Akt, and factors related to apoptotic modulation. In comparison to the control group, NEC induction resulted in a significant reduction in MUC1 expression, accompanied by an elevation in enterocyte apoptosis. In NEC and Si-MUC1 Caco-2 cells, downregulation of PI3K/AKT signals and Bcl-2 was observed, while upregulation of Bax, CytoC, and Caspase 3 at both mRNA and protein levels was prominent. Retinoic acid supplementation exhibited a noteworthy increase in MUC1, AKT, and Bcl-2 mRNA and protein expressions, coupled with a decrease in Bax, CytoC, and Caspase 3, thereby mitigating apoptosis in NEC. Our findings suggested that reduced MUC1 expression in NEC contributes to the upregulation of enterocyte mitochondrial apoptosis through the PI3K/AKT signaling pathway. Retinoic acid supplementation emerges as a potential therapeutic strategy for NEC, demonstrating its ability to upregulate MUC1 expression and attenuate apoptosis via the PI3K/AKT signaling pathway.

Predicting the evolution from first-episode psychosis to mood or psychotic disorder: A systematic review of biological markers.

The development of paraclinical tools to assist clinical assessment is already widespread in nearly all other medical specialties. In psychiatry, many efforts are being made to improve management strategies using these new techniques. The first episode psychosis (FEP) is a clinical entity whose evolution after onset is difficult to predict in the current state of our practices. Our main objective was to identify from the literature the most promising biological markers for early prediction of thymic or psychotic trajectories following FEP. We performed a systematic literature review on 4 databases: PubMed, Scopus, PsycINFO, and Web of Science following PRISMA guidelines and using search terms related to FEP and biomarkers. Eight studies were included in our final analysis. Several biomarkers showed promising discriminatory capacities for predicting post-FEP evolution: the interleukins IL-6, IL-12p40, IL-1β, and the mRNA expression levels of the DICER-1 and AKT-1 genes. Other studies that opted for broad-spectrum strategies also highlighted new leads for the discovery of additional biomarkers. Overall, our results indicate the value of replicating studies targeting the analysis of the predictive capacities of several biological markers. It also appears important to homogenize methodologies and favor the construction of predictive models on several of these markers to reinforce their statistical significance.

Anticancer Effectivity of Nanocrystals Derived from Mangosteen (Garcinia mangostana) Peel Extract on Leukemia HL-60 Cells

Leukemia, characterized by abnormal leukocyte proliferation, ranks ninth in Indonesia as the most common cancer. While treatments such as chemotherapy and radiation effectively target cancer cells, they also risk damaging healthy blood cells. This has spurred interest in exploring low-toxicity herbal compounds as potential therapies, with mangosteen peel emerging as a widely researched option. Nanotechnology, which has the potential to enhance the bioavailability of herbal compounds, is also a focus of extensive research. This study objective was to assess the impact of Mangosteen Peel Nanocrystal (MPN) on HL-60 leukemia cells by analyzing various parameters, including cytotoxicity, reactive oxygen species (ROS) levels, senescence, and gene expression changes. MPN was prepared with high-speed milling and characterized using particle size analyzers, microscopy, and stability assessments. HL-60 cells were cultured and subjected to MPN treatment. Cytotoxicity was evaluated using WST-8 assays, ROS levels were assessed using flow cytometry, and senescence analyses using Senescence-Associated b-Galactosidase Staining. AKT and FLT-1 gene expression were determined via qRT-PCR. MPN has been successfully characterized as a nanoparticle based on size, stability, and morphology. MPN has an impact on leukemia cells by increasing cytotoxicity, decreasing ROS levels, inducing senescence, and modulating AKT and FLT-1 gene expressions. The findings suggest potential implications for MPN in targeting leukemia cells. The study sheds light on the promising effects of MPN in leukemia cell models, indicating its potential applications in targeting cancer cells, inducing senescence, decreasing ROS levels, and modulating gene expressions related to cell survival and proliferation.

A Case of NTRK Fusion Corpus Sarcoma With Pseudobiphasic Growth Pattern and Literature Review.

The incidence of neurotrophic tyrosine kinase receptor (NTRK) fusion uterine sarcoma is extremely low, and reports have been mostly focused on cases localized to the cervix. So far, only 4 cases have been reported of the uterine corpus. In this study, we reported a case of NTRK fusion corpus sarcoma. This study aimed to expand the morphologic spectrum of this tumor, which showed adenosarcoma-like features not previously described. The tumor was confined to the uterine corpus, polypoid growth, comprised predominantly of a fascicular proliferation of spindle cells, entrapping benign endometrial glands, and exhibited a pseudo-biphasic growth pattern. The tumor showed coexpression of S-100, CD34, and pan-Trk by immunohistochemistry, DNA-sequencing identified TPR-NTRK1 gene fusion and AKT1(E17K) mutation. Four cases of NTRK fusion corpus sarcoma were reviewed. The clinicopathologic features, immunohistochemical phenotype, molecular testing, and prognosis of 5 cases including this one were summarized and analyzed. Most cases exhibited an infiltrative g rowth pattern and showed mild or moderate cytologic atypia. The potential for these tumors to be misclassified as uterine adenosarcoma or other uterine mesenchymal tumors. The diagnosis relies on pan-Trk, S-100, CD34 immunohistochemistry, and molecular testing. Surgical resection is the mainstay of treatment for most patients. Distinguishing these tumors from morphologic mimics is significant because patients with advanced-stage disease may be treated with TRK inhibitors.

Plasma mutation profile of precursor lesions and colorectal cancer using the Oncomine Colon cfDNA Assay

BackgroundColorectal cancer (CRC) is the second leading cause of cancer death worldwide. Early detection of precursor lesions or early-stage cancer could hamper cancer development or improve survival rates. Liquid biopsy, which detects tumor biomarkers, such as mutations, in blood, is a promising avenue for cancer screening.AimTo assess the presence of genetic variants in plasma cell-free tumor DNA from patients with precursor lesions and colorectal cancer using the commercial Oncomine Colon cfDNA Assay.Material and methodsCell-free DNA (cfDNA) samples from the plasma of 52 Brazilian patients were analyzed. Eight patients did not have any significant lesions (five normal colonoscopies and three hyperplastic polyps), 24 exhibited precursor lesions (13 nonadvanced adenomas, 10 advanced adenomas, and one sessile serrated lesion), and 20 patients with cancer (CRC). The mutation profile of 14 CRC-associated genes were determined by next-generation sequencing (NGS) using the Oncomine Colon cfDNA Assay in the Ion Torrent PGM/S5 sequencer.ResultsThirty-three variants were detected in eight genes (TP53, PIK3CA, FBXW7, APC, BRAF, GNAS, KRAS, and SMAD4). No variants were detected in the AKT1, CTNNB1, EGFR, ERBB2, MAP2K1 and NRAS genes. All variants were considered pathogenic and classified as missense or truncating. The TP53 gene harbored the most variants (48.48%), followed by the KRAS gene (15.15%) and the APC gene (9.09%). It was possible to detect the presence of at least one pathogenic variant in cfDNA in 60% of CRC patients (12/20) and 25% of precursor lesions (6/24), which included variants in three patients with nonadvanced adenoma (3/13 – 23.08%) and three with advanced adenomas (3/10 – 30%). No variants were detected in the eight patients with normal findings during colonoscopy. The detection of mutations showed a sensitivity of 60% and a specificity of 100% for detecting CRC and a sensitivity of 50% and a specificity of 100% for detecting advanced lesions.ConclusionThe detection of plasma NGS-identified mutations could assist in early screening and diagnostic of CRC in a noninvasive manner.

Rotavirus-Inspired Nanointerface Engineered Biosensors for All-in-One Cancer Diagnosis.

Ultrasensitive and population-scale cancer screening technologies are critical to reducing cancer mortality. However, the current qRT-PCR falls short in high-throughput screening of multiple cancers. Here, a rotavirus-inspired multicancer diagnosis system (RMDS) is developed via nanointerface engineering. RMDS employs Y-shaped DNA (YDNA) probes to encircle the graphene quantum dots (GQDs) for nanointerface modification. The biotransduction mechanisms at the nanointerface are systematically investigated. RMDS greatly enhances the transduction efficiency of biological analytes by optimizing the probe density and configuration. RMDS realizes ultrasensitive detection of the lung cancer KARS G12D mutation with a limit of detection (LoD) of 5.7 aM and the breast cancer-related AKT2 gene (LoD: 3.0 aM). The multiple chambers enable simultaneous diagnosis of multiple cancers and determination of cancer progression. Clinical validation shows RMDS can be a practical solution, which could complement or replace qRT-PCR and become the next-generation all-in-one tool for large-scale population cancer screening.

The niosomal nelarabine as a promising nano combination for retinoblastoma treatment: an in vitro study—experimental research

Introduction: Retinoblastoma (RB), the most commonly occurring intraocular malignancy among children globally, represents 3% of childhood cancers. In the current study, the authors aim to evaluate the effectiveness of a new formulation of nelarabine (niosomal nelarabine) on RB cancer cells. Methods: Field emission scanning electron microscopy (FE-SEM) and dynamic light scattering (DLS) characterized the physical properties of nelarabine nanoparticles. After cultivation of the Y79 cell line, the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test was performed to determine IC50 of niosomal nelarabine (Nio-Nelarobine) and also the cytotoxicity of Nio-Nelarobine and doxorubicin against Y79 cell line was investigated. The level of apoptosis was assessed by flow cytometry in selected groups. Also, the PTEN/AKT/FOXO1 gene expression level was measured using qRT-PCR. Results: Y79 cell lines were treated with Nio-Nelarobine and doxorubicin. The treatment resulted in a dose-dependent inhibition of Y79 cell viability. However, Nio-Nelarobine showed a higher inhibitory activity with a diameter of about 167 nm. Both Nio-Nelarobine and doxorubicin induced apoptosis in cells, but Nio-Nelarobine treatment resulted in a higher number of apoptotic cells than doxorubicin treatment. The qRT-PCR results showed that the treatment with Nio-Nelarobine and doxorubicin led to an increase in the expression of PTEN and FOXO1 genes, while decreasing the expression of the AKT gene. Furthermore, the statistical significance of these results was higher in the Nio-Nelarobine group than in the doxorubicin group. Conclusions: Nio-Nelarobine may be a functional therapeutic combination for RB treatment. Further experimental and preclinical investigations are necessary to verify this impact in greater detail.

Identification of key genes related to growth of largemouth bass (Micropterus salmoides) based on comprehensive transcriptome analysis.

Largemouth bass is an economically important farmed freshwater fish species that has delicious meat, no intermuscular thorns, and rapid growth rates. However, the molecular regulatory mechanisms underlying the different growth and developmental stages of this fish have not been reported. In this study, we performed histological and transcriptomic analyses on the brain and dorsal muscles of largemouth bass at different growth periods. The brain and muscle tissue were dehydrated, embedded, sliced and stained with hematoxylin-eosin. Images were captured under a microscope and acquired using a microphotographic system. Differential expression between groups was analyzed using DESeq2. GO functional analysis and KEGG pathway analysis were then performed for differentially expressed genes. RT-qPCR validates the reliability of transcriptome sequencing data. Smaller fish had more new muscle fiber numbers and wider intermuscular spaces compared to big specimens. Axons and nerve fibers were more pronounced in the telencephalons of big fish than in small fish. A total of 19,225 differentially expressed genes (DEGs) were detected in the muscle tissue, among which 7,724 were upregulated and 11,501 were downregulated, while a total of 5,373 DEGs were detected in the brain, among which 2,923 were upregulated and 2,450 were downregulated. GO and KEGG enrichment analyses indicated that nucleic acid binding, cytoskeletal motor activity, DNA binding, circadian rhythm, glycolysis/gluconeogenesis, and osteoclast differentiation were related to brain development while binding, cytoskeletal protein binding, biological processes, c-type lectin receptors, mitogen-activated protein kinase (MAPK) signaling pathways, and osteoclast differentiation were related to muscle growth. Stat3, pparg, akt1, mapk3, and mapk1 genes were mainly involved in the growth and development of largemouth bass. These results provide novel perspectives for deepening our understanding of the mechanisms underlying the growth and development and performing genetic selection in largemouth bass.

Phloretin-nanospanlastics for targeting the Akt/PI3K signaling pathways in dimethylhydrazine-induced colon cancer in mice.

Colorectal cancer is the third most common cancer worldwide, accounting for approximately 10% of all cancer cases. It is also the second leading cause of cancer-related deaths globally. Phloretin is a natural compound found in apples and other fruits. It has been studied for its potential health benefits, including antioxidant and anti-inflammatory properties. However, more research is needed to fully understand its impact on cancer prevention or treatment. This article aimed to prepare phloretin-nanospanlastics (Ph-NSLs) to evaluate their effects on dimethylhydrazine (DMH)-induced colon cancer in mice. Morphology, Particle size, zeta potential, UV-vis, entrapment efficiency, polydispersity index, FT-IR spectra, and drug release of phloretin and Ph-NSLs at pH6.8.were described. Ph-NSLs were also tested for their anti-cancer properties in DMH-induced colon cancer in mice. A 36 mice were divided into 6 groups; Normal control, DMH (20mg/k.g.b.w.), DMH+Ph-NSLs (25mg/k.g.b.w.), DMH+Ph-NSLs (50mg/k.g.b.w.), DMH+5-FU(20mg/k.g.b.w.), DMH+Ph-NSLs (50mg), 5-FU (20mg). Ph-NSLs were tested for their anticancer properties in DMH-treated mice by evaluating the IC50, viability and inhibitory values of Ph-NSLs against Caco-2. Also, the effect of Ph-NSLs administration on number of surviving mice, number of tumors/mice, average of tumor size, Hb, RBCs, WBCs, C19-9, MDA, GSH, SOD, IL-2, TNF-α, TGF-β1, CEA, and P53 levels in mice treated DMH were estimated. The synthesized Ph-NSLs were uniform, spherically shaped, and well dispersed, with a size, entrapment efficiency, and polydispersity index of approximately 114.06±8.35nm, 78.60%, and 0.05, respectively. The zeta potential value of Ph-NSLs was measured at -21.5±1.47mV. Zeta potential reflects the surface charge of nanoparticles and affects their stability and interactions. UV spectra of phloretin and Ph-NSLs showed strong absorption peaks at 225 and 285nm. These peaks correspond to specific wavelengths where the compound absorbs light. The percentage of Ph- NSLs release was found to be 56.87±2.45%. IC50 of Ph-NSLs was recorded 15.76±0.42μg/ml and the viability and inhibitory values of Ph-NSLs against Caco-2 cell lines was resorded 2.39, and 97.61%, respectively at 100μg/ml as well as 10.3, and 89.7%, respectively at 50μg/ml.Moreover, The combination of 5-FU and Ph-NSLs resulted in a moderate increase in survival and significantly reduces tumor size and number, showing enhanced anticancer efficacy compared to individual treatments as well as attenuated levels of hemoglobin (Hb), red blood cells (RBCs), and white blood cells (WBCs). Reduced plasma cancer antigen 19-9 (CA19-9) levels as well as improved of colon malondialdehyde (MDA), reduced glutathione (GSH), superoxide dismutase (SOD), interleukine-2 (IL-2), tumor necrosis factor-alpha (TNF-α), tumor growth factor-beta1 (TGF-β1), carcinoembryonic antigen (CEA), and tumor protein (P53) levels. Also, Ph-NSLs and 5FU, either alone or together, decreased the expression of the Akt and PI3K genes in the colon. The combination of Ph-NSLs and 5FU showed more pronounced anticancer activity than Ph-NSLs administered individually. The combination of 5-FU and Ph-NSLs significantly enhances anticancer efficacy, reducing both the number of tumors and average tumor size more effectively than either treatment alone. This synergistic effect leverages 5-FU's inhibition of DNA synthesis and phloretin's induction of apoptosis and inhibition of cell proliferation, offering a promising approach for improved cancer treatment outcomes.

Cytotoxic Impact of Naringenin-Loaded Solid Lipid Nanoparticles on RIN5F Pancreatic β Cells via Autophagy Blockage.

Autophagy plays a crucial role in modulating the proliferation of cancer diseases. However, the application of Naringenin (Nar), a compound with potential benefits against these diseases, has been limited due to its poor solubility and bioavailability. This study aimed to develop solid lipid nanoparticles (Nar-SLNs) loaded with Nar to enhance their therapeutic impact. In vitro experiments using Rin-5F cells exposed to Nar and Nar-SLNs were carried out to investigate the protective effects of Nar and its nanoformulation against the pancreatic cancer cell line of Rin-5F. Treatment with Nar and Nar-SLN led to an increase in autophagic markers (Akt, LC3, Beclin1, and ATG genes) and a decrease in the level of miR-21. Both Nar and Nar-SLN treatments inhibited cell proliferation and reduced the expression of autophagic markers. Notably, Nar-SLNs exhibited greater efficacy compared to free Nar. These findings suggest that SLNs effectively enhance the cytotoxic impact of Nar, making Nar-SLNs a promising candidate for suppressing or preventing Rin-5F cell growth.

Association of SIGLEC9 Expression with Cytokine Expression, Tumor Grading, KRAS, NRAS, BRAF, PIK3CA, AKT Gene Mutations, and MSI Status in Colorectal Cancer.

SIGLEC9 (sialic acid-binding Ig-like lectin 9) is a molecule thought to have a significant influence on the immune properties of the colorectal cancer (CRC) tumor microenvironment (TME). In our study, we assessed the expression of the SIGLEC9 protein in CRC tissue and the surgical margin tissue. Using RT-PCR, we analyzed mutations in the KRAS, NRAS, BRAF, PIK3CA, and AKT genes. We observed a significantly elevated expression of the SIGLEC9 protein in CRC tissue compared to the control group. No significant differences were observed in SIGLEC9 protein expression depending on mutations in the KRAS, NRAS, BRAF, PIK3CA, and AKT genes or microsatellite instability (MSI) status. However, we found a significantly higher expression of the SIGLEC9 protein in high-grade tumors compared to the low-grade tumors group. SIGLEC9 expression was significantly associated with the expression of multiple cytokines, chemokines, and growth factors in the CRC TME. These associations suggest the significant potential of SIGLEC9 as a molecule that plays a crucial role in shaping the immune properties of the CRC TME, as well as its potential therapeutic relevance, particularly in the group of high-grade CRC tumors.

Analysis of the potential long-lasting effects of probiotic Debaryomyces hansenii CBS 8339 on trained immunity in newborn goats

Trained immunity has been described as the memory capacity of the innate immune system. Several microbial components have been shown to induce trained immunity. Research on the potential of probiotics to trigger these effects has been limited to a few in vitro studies but remains completely unknown in vivo. Components from the probiotic Debaryomyces hansenii CBS 8339 (Dh) have been shown to induce innate immune memory in goat kids and calves. In the present study, stimulating innate immune cells from newborn goats with probiotic Dh increased respiratory burst activity and nitric oxide production, while cell phagocytosis was unaffected. Glucose uptake was enhanced in goat's cells stimulated with Dh, but lactate production was decreased. In newborn goats, after the training scheme (via oral probiotic administration), cell phagocytosis, nitric oxide production and glycolysis — through the upregulation of AKT and HIF1A gene expression, glucose consumption and lactate production— were enhanced. The expression of IL1B gene was similar between the D. hansenii and control groups. Moreover, the potential long-lasting effects were assessed 30 days after initiation of the training scheme. Cell phagocytosis, respiratory burst and myeloperoxidase activity were enhanced, while glycolytic parameters remained unaffected. Altogether, the results of the present study suggest that the immune training scheme may induce trained immunity by the probiotic D. hansenii in newborn goats. However, our findings were not conclusive regarding the long-lasting (one-month) effects of trained immunity by probiotics.

Akt regulates the fertility of Coridius chinensis by insulin signaling pathway

Akt (also known as protein kinase B) belongs to the multifunctional serine/threonine kinase family and is an important component of the insulin signaling pathway that plays a key role in many biological processes such as cell growth, proliferation, and survival. However, few studies have reported the effect of Akt on reproduction in Hemiptera. In this study, we cloned and characterized the Akt gene from Coridius chinensis (CcAkt). The open reading frame of CcAkt has a length of 1,563 bp and encodes 520 amino acids. It has a conserved pleckstrin homology domain (PH), catalytic domain of serine/threonine protein kinases (S_TKc), and extension of Ser/Thr-type protein kinases (S_TK_X). Phylogenetic analysis showed that CcAkt and HhAkt of Halyomorpha halys had the highest similarity. Analysis of temporal and spatial expression patterns revealed that CcAkt is expressed throughout development and in various tissues of C. chinensis adults. CcAkt was highly expressed in the female adult and the fourth-instar nymph, as well as in the testis and ovary of C. chinensis. Injection of bovine insulin and methoprene induced the CcAkt expression, whereas that of 20-hydroxyecdysone significantly reduced the CcAkt expression. These three hormones, however, induced the expression of vitellogenin (Vg) and vitellogenin receptor (VgR). In unmated females, knockdown of CcAkt resulted in decreased expression of CcVg and CcVgR, stunted the development of the ovarioles, decreased the number of eggs and hatching rate. These findings from RNA interference experiment suggested that CcAkt may be involved in regulating the reproduction of C. chinensis.

The Impact of Acute Myeloblastic Leukemia-Derived Small Extracellular Vesicles on the Bone Marrow Mesenchymal Stem Cells: Expression of G1 Cell Cycle Progression Genes

Background: Leukemia progression is intricately linked to interactions with neighboring cells within the bone marrow microenvironment (BMM), and small extracellular vesicles (sEVs) emerge as vital mediators in facilitating these interactions. Objectives: This study examined the proliferation effects of sEVs-derived from acute myeloid leukemia (AML) in the HL60 cell line on the cell cycle progression of bone marrow mesenchymal stem cells (BM-MSCs), a key element of the BMM. Methods: Small extracellular vesicles were isolated from the HL60 cell line supernatant, using the ExoCib kit, and characterized through flow cytometry, transmission electron microscopy (TEM), dynamic light scattering (DLS), and bicinchoninic acid (BCA) protein assay. bone marrow mesenchymal stem cells were cultured and treated with various concentrations (20 μg/mL, 50 μg/mL, and 80 μg/mL) of AML-derived sEVs for 24, 48, and 72 hours. The effects on cell proliferation and viability were assessed, using methylthiazole tetrazolium test (MTT) and Ki-67 assays, while cell cycle progression and apoptosis were analyzed via flow cytometry. RT-qPCR was performed to evaluate the expression levels of CCND1, CDK4, CDK6, and AKT1 genes. Results: The proliferation effects of AML-derived sEVs on BM-MSCs were both dose- and time-dependent, with optimal effects observed at 50 μg/mL after 48 hours. Flow cytometry analysis revealed a significant increase in the G1 phase, showing a 1.8-fold change compared to the control group (P < 0.0001). RT-qPCR results demonstrated a significant upregulation of CCND1 (3.5-fold, P < 0.0001), CDK4 (3.2-fold, P < 0.0001), CDK6 (2.5-fold, P < 0.0001), and AKT1 (3.2-fold, P < 0.0001) expression levels, along with increased Ki-67 (2.3-fold, P < 0.0001) levels. Moreover, treatment with 50 μg/mL of AML-derived sEVs resulted in a notable reduction in BM-MSC apoptosis (0.57-fold decrease, P < 0.0001). Conclusions: Our findings revealed that AML extracellular vesicles could alter the gene expression associated with the proliferation of BMSCs to increase their proliferation by stimulating the cell cycle of MSCs through the G1 phase.

Synergistic anticancer efficacy of polydatin and sorafenib against the MCF-7 breast cancer cell line via inhibiting of PI3K/AKT/mTOR pathway and reducing resistance to treatment

Polydatin (PD), a glucoside derivative of resveratrol, has been investigated for its potential to mitigate sorafenib (SOF) side effects and combat multidrug resistance in cancer treatment. The study evaluated its mechanism of action for inhibiting the protein kinase B/mTOR pathway in promoting breast cancer proliferation. The combined PD and SOF have synergistic effects with a combination index (CI) < 1 in the liver (HepG2) and breast (MCF-7) cancer cell lines. Molecular docking studies were conducted to analyze interactions of PD& SOF with protein kinases as well as apoptotic and multidrug resistance proteins, including AKT1, PI3K, mTOR, Apaf-1, and ABCB1 in MCF-7 cells. Experimental validation through real-time PCR confirmed. PD has a strong binding affinity, particularly with AKT1 (−56 kcal/mol) and ABCB1 (−27.16 kcal/mol), a gene associated with multidrug resistance. These interactions were linked to anti-proliferative anti-angiogenic effects and reduced resistance to treatment, demonstrating PD has potential therapeutic benefits. Furthermore, PD combined with SOF induced apoptosis, inhibited cell growth, and arrested MCF-7 cells in the sub-G1 phase with increased intracellular ROS. This was accompanied by reduced expression of AKT1 and ABCB1 genes, reinforcing the anticancer efficacy of PD/SOF combination therapy. In conclusion, the findings suggest that PD/SOF could serve as a promising anticancer treatment strategy, warranting further investigation for potential clinical applications and mechanistic studies in vivo.

Proteus Syndrome: Description of Two Clinical Cases

Background. Proteus syndrome is extremely rare congenital multisystem disease with high variability in clinical manifestations. Its prevalence is unknown, there are less than 200 cases in the world literature. The syndrome is a classic example of somatic mosaicism, and all target drugs for its management are based on it. Clinical case description. This article describes two clinical cases with somatic variants of the nucleotide sequence in the AKT1 gene, mosaic form, revealed by the NGS method. Target drug (mTOR-inhibitors group) was assigned in one case. Conclusion. The description of the phenotypic features of patients with Proteus syndrome is crucial as this pathology is very rare. It is necessary to increase the awareness of clinicians about this disease to develop a plan for dynamic follow-up with consideration to life-threatening complications (malignant tumors and thrombembolia risk). Genetic verification of Proteus syndrome is mandatory nowadays as target therapy is actively developed and implemented, thus, revision of clinical guidelines is recommended.

Investigating the Mechanisms of Anti-tumoral Effect of Combination Therapy of Calcitriol and Sodium Pentaborate Pentahydrate on HepG2 Hepatocellular Carcinoma Cells.

Hepatocellular carcinoma (HCC) is one of the most common primary liver cancers worldwide and is often associated with poor prognosis due to drug resistance. Combination therapies demonstrate superior efficacy at lower drug dosages on cancer cells compared to single treatments, resulting in less drug resistance in the cells. This study investigates the synergistic anti-tumoral effects of calcitriol, the biologically active form of vitamin D, and sodium pentaborate pentahydrate (NaB) on HepG2 cells. We examined the cell viability of NaB, calcitriol, or the combination of NaB and calcitriol on HepG2 cells and healthy human hepatic stellate cells (HHSC) using MTS. Our findings showed that combination therapy with 3.3mM NaB and 1µM calcitriol has a synergistic effect and a more cytotoxic effect on HepG2 cells. This combination significantly increased apoptosis and ROS levels compared to treatment alone with NaB or calcitriol. Gene expression and proteomics analysis revealed inhibition of DNA replication and the cell cycle process, further confirming the potent anti-proliferative effects of the combination therapy. When HepG2 cells were treated with a combination of 3.3mM NaB and 1µM calcitriol, mRNA levels of apoptosis-related genes AKT1 and MDM2 were downregulated, while p53 was upregulated. Additionally, cell cycle-related genes CDKN1A, GADD45A, and p27 were upregulated, whereas MCM2, MCM5, and MCM7 were downregulated. Furthermore, genes associated with the vitamin D receptor (VDR), including VDR and CYP24A1, were upregulated, while CYP27B1 was downregulated. Our proteomic analysis revealed decreased MCM2 and MCM5 protein expressions which was confirmed by western blotting. In conclusion, this study highlights the potential of NaB and calcitriol as a promising therapeutic strategy for HCC.

Regulation of pro-apoptotic and anti-apoptotic factors in obesity-related esophageal adenocarcinoma

BackgroundObesity is a risk factor for esophageal adenocarcinoma (EAC). It was reported that obesity -associated inflammation correlates with insulin resistance and increased risk of EAC. The objective of the study is to investigate the role of obesity associated inflammatory mediators in the development of EAC.MethodsWe included 23 obese and nonobese patients with EAC or with or without Barrett’s esophagus (BE) after IRB approval. We collected 23 normal, 10 BE, and 19 EAC tissue samples from endoscopy or esophagectomy. The samples were analyzed for the expression levels of pro-apoptotic and anti-apoptotic factors, PKC-δ, cIAP2, FLIP, IGF-1, Akt, NF-kB and Ki67 by immunofluorescence and RT-PCR. We compared the expression levels between normal, BE, and EAC tissue using Students’ t-test between two groups.ResultsOur results showed decreased gene and protein expression of pro-apoptotic factors (bad, bak and bax) and increased expression of anti-apoptotic factors (bcl-2, Bcl-xL) in BE and EAC compared to normal tissues. There was increased gene and protein expression of PKC-δ, cIAP2, FLIP, NF-kB, IGF-1, Akt, and Ki67 in BE and EAC samples compared to normal esophagus. Further, an increased folds changes in mRNA expression of proapoptotic factors, antiapoptotic factors, PKC-δ, IGF-1, Akt, and Ki-67 was associated with obesity.ConclusionPatients with EAC had increased expression of cIAP2 and FLIP, and PKC-δ which is associated with inhibition of apoptosis and possible progression of esophageal adenocarcinoma.

Shengqiyichang decoction regulates antitumor immunity in colorectal cancer by downregulating lymphocyte antigen 6 family member G6D via the protein kinase B/p38 mitogen-activated protein kinase signaling pathway

The traditional Chinese medicine (TCM) formulation Shengqiyichang Decoction (SQYCD) has been reported to stimulate host immunity, and it has been administered for the treatment of colorectal cancer (CRC). Here, we applied network and bioinformatics analyses to elucidate the mechanisms by which SQYCD ameliorates CRC and validated its modes of action via in vivo and in vitro experiments. We identified 46 active compounds in SQYCD and selected 237 proteins as potential therapeutic targets in CRC, most notably p38 mitogen-activated protein kinase (p38⍺). Bioinformatics analyses demonstrated differential expression in CRC tissues and prognostic value of the genes encoding TNFα, MAPK14, CASP-3, MAPK1, AKT1, PRKACA, VEGF, IL-6, EGFR and ESR1. We then plotted receiver operating curves (ROC) and time-ROC for the differentially expressed genes (DEGs) ESR1 and AKT1 to predict the progress of CRC. We speculated that the AKT/p38α-MAPK signaling pathway is associated with the clinical prognosis of CRC. In a mouse model, we found that SQYCD inhibits CRC tumor growth by increasing CD4+ and CD8+ T cell abundance and decreasing the ratio of T-regulatory cells (Tregs) in the tumor microenvironment. In cultured mouse CRC cells, SQYCD selectively upregulated levels of the CRC-associated protein lymphocyte antigen 6 family member G6D, while the AKT activator SC-79 reversed this effect. The discoveries made herein suggest that SQYCD exerts a therapeutic effect in CRC by inhibiting Treg recruitment via inhibition of the AKT/p38α/LY6G6D signaling axis.

Genetic signatures of AKT1 variants associated with worse COVID-19 outcomes - a multicentric observational study.

The COVID-19, triggered by the SARS-CoV-2 virus, has varied clinical manifestations, ranging from mild cases to severe forms such as fatal pneumonia and acute respiratory distress syndrome (ARDS). Disease severity is influenced by an exacerbated immune response, characterized by high pro-inflammatory cytokine levels. Inhibition of AKT can potentially suppress pathological inflammation, cytokine storm and platelet activation associated with COVID-19. In this study, we aimed to investigate the rs2494746 and rs1130214 variants in the AKT1 gene associated with severe COVID-19 outcomes. Peripheral blood samples and sociodemographic data from 508 individuals with COVID-19, measuring plasma cytokine concentrations using ELISA and genotyped the AKT1 variants. The rs2494746-C allele was associated with severity, ICU admission, and death from COVID-19. The C allele at rs1130214 was linked to increased TNF and D-dimer levels. Moreover, both variants exhibited an increased cumulative risk of disease severity, ICU admission, and mortality caused by COVID-19. In the predictive analysis, the rs2494746 obtained an accuracy of 71%, suggesting a high probability of the test determining the severity of the disease. Our findings contribute to understanding the influence of the AKT1 gene variants on the immunological damage in individuals infected with SARS-CoV-2.