Upregulation Of Cell Proliferation Research Articles

DGAT2 Plays a Crucial Role to Control ESRRAPROX1 Transcriptional Network to Maintain Hepatic Mitochondrial Sustainability.

Diacylglycerol O-acyltransferase 2 (DGAT2) synthesizes triacylglycerol (TG) from diacylglycerol; therefore, DGAT2 is considered as a therapeutic target for steatosis. However, the consequence of inhibiting DGAT2 is not fully investigated due to side effects including lethality and lipotoxicity. In this article, we observed the role of DGAT2 in hepatocarcinoma. The role of DGAT2 is analyzed via loss-of-function assay. DGAT2 knockdown (KD) and inhibitor treatment on HepG2 cell line was analyzed. Cumulative analysis of cell metabolism with bioinformatic data were assessed, and further compared with different cohorts of liver cancer patients and non-alcoholic fatty liver disease (NAFLD) patients to elucidate how DGAT2 is regulating cancer metabolism. Mitochondrial function is suppressed in DGAT2 KD HepG2 cell along with the decreased lipid droplets. In the aspect of the cancer, DGAT2 KD upregulates cell proliferation. Analyzing transcriptome of NAFLD and hepatocellular carcinoma (HCC) patients highlights negatively correlating expression patterns of 73 lipid-associated genes including DGAT2. Cancer patients with the lower DGAT2 expression face lower survival rate. DGAT2 KD cell and patients' transcriptome show downregulation in estrogen- related receptor alpha (ESRRA) via integrated system for motif activity response analysis (ISMARA), with increased dimerization with corepressor prospero homeobox 1 (PROX1). DGAT2 sustains the stability of mitochondria in hepatoma via suppressing ESRRA-PROX1 transcriptional network and hinders HCC from shifting towards glycolytic metabolism, which lowers cell proliferation.

304 NF2 Loss-of-Function and Hypoxia Together Drive Radioresistance in Grade 2 Meningiomas

INTRODUCTION: Grade 2 meningiomas (G2Ms) are associated with an aggressive natural history characterized by recurrence and resistance to radiation therapy. Histopathological necrosis is associated with worse progression-free survival (PFS) following radiation therapy in G2Ms. We seek to understand the molecular pathophysiology of radiation resistance and to identify molecular biomarkers and therapeutic targets for radioresistant G2M. METHODS: We performed genetic sequencing and histopathological analysis of 121 G2Ms. Using the IOMM-Lee meningioma cell line, we investigated the role of tumor hypoxia and genes of interest in radio-resistance through lentivirally targeted gene knockdown and clonogenic survival assays following exposure to ionizing radiation. Transcriptional profiling of cells in in vitro and in 38 patient G2Ms was performed using RNA sequencing. RESULTS: NF2 loss of function mutations were significantly associated with tumor necrosis in G2Ms (p = 0.0026). Patients with NF2mutation and spontaneous necrosis had worse post-radiation PFS compared to NF2 wildtype patients without necrosis (p = 0.035). In the setting of hypoxia, NF2 knockdown significantly increased radio-resistance in vitro (p < 0.001). Gene set enrichment analysis on in vivo and in vitro data and functional assays revealed downregulation of apoptosis and upregulation of cell proliferation in NF2-deficient and necrotic group compared to control. Integration of differentially expressed genes in both NF2-deficient cells in vitro and human necrotic tumors identified shared upregulation of HES1, XDH, NOG, IGFBP3, ALOXE3, THSB1, and TRIML2, suggesting these genes may represent targets for radio-sensitization or biomarkers of radio-resistance. CONCLUSIONS: NF2 loss-of-function in the setting of hypoxia confers radiation resistance through a transcriptional program that promotes proliferation and DNA repair. Targeting molecular pathways identified by transcriptional profiling may improve local control in patients with radioresistant G2M.

Abstract 98: SETDB1 promotes tumorigenesis in endometrial cancer through suppressing anti-tumor immune response and increasing proliferation

Abstract Background: An H3K9 methyltransferase and cancer driver gene, SETDB1, promotes tumorigenesis in many cancer types, such as melanoma, and breast cancer. SETDB1 is often overexpressed and correlates with worse survival. Multiple mechanisms have been reported for its oncogenesis from promoting tumorigenesis and regulating immune evasion. The roles of SETDB1 in driving endometrial cancer (EC) progression is not well-studied. Methods: EC cell lines were used for knockout SETDB1 using CRISPR-Cas9 system, RNA-seq was conducted to discover SETDB1 regulated genes. ChIP-qPCR was performed to identify SETDB1 direct targets. Immunocompromised NSG and immunocompetent C57BL/6 mice were utilized to investigate SETDB1 promoted tumor growth, immune evasion, and mice survival. IHC and flow cytometry was conducted to evaluate macrophage infiltration. Cytokines were screened by qPCR for potential macrophage chemoattractant. Results: In vitro, knockout SETDB1 decreased cell proliferation and enhanced G1/S cell cycle arrest. RNA-seq data revealed that in SETDB1−/− clones, many important oncogenes such as POLR2A, MUC5AC, and MKI67 were downregulated, while tumor suppressors such as PGR, RERG, and ZNF582 were upregulated. ChIP assay discovered that SETDB1 directly binds and represses ZNF582, ZNF266, ZNF841 expression via depositing H3K9me3 at promoter region. In vivo, SETDB1−/- significantly reduced tumor growth and prolonged NSG mice survival up to 100 days. SETDB1 promoted tumor growth via 1. Intrinsic: increasing mitotic figure, pH3-S10 and Ki67 level, 2. Extrinsic: blocking macrophage infiltration and chemokine secretion. Our in vitro functionality assay confirmed that bone-marrow derived M1-polarized macrophages promoted greater killing for SETDB1−/− cells. We surveyed 17 cytokines and found CCL5, CXCL9, and IL6 mRNA expression were enhanced in SETDB1−/− tumors suggesting active immune response. Consistent with the elevated CCL5 mRNA expression, CCL5 protein level was also upregulated in macrophages and cancer cells in SETDB1−/- tumors. To prove SETDB1 directly regulates CCL5 expression, we re-expressed SETDB1 in SETDB1−/− cells, and indeed, the SETDB1 re-expression did suppress CCL5 expression. Using immunocompetent C57BL/6 mice, we further confirmed that SETDB1 promotes tumor growth of MSH2-369 mice cells and blocks macrophage infiltration. To understand SETDB1 regulated pathways in EC patients’ samples, using EC-TCGA data, we discovered that high SETDB1 expression correlated with higher level of active transcription genes and reduced immune response genes. Our findings provide the mechanisms behind these observations. Conclusion: SETDB1 drives EC tumorigenesis by both intrinsically upregulating cell proliferation genes and promoting immune escape extrinsically. Our novel findings suggest SETDB1 is a promising anticancer target for EC patients. Citation Format: Kiarash Salari, Jiaqing Hao, Matthew Wells, Eleanor Johnson, Ryan Jilek, Jack Gilbert, Ryan Mclerran, David Meyerholz, Melinda Yates, Bing Li, Shujie Yang. SETDB1 promotes tumorigenesis in endometrial cancer through suppressing anti-tumor immune response and increasing proliferation [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 98.

Lipopolysaccharide induces CCL2 through TLR4 signaling and promotes esophageal squamous cell carcinoma cell proliferation.

Poor oral health is an independent risk factor for upper-aerodigestive tract cancers, including esophageal squamous cell carcinoma (ESCC). Our previous findings suggest that high expression of toll-like receptor (TLR) 4, which recognizes lipopolysaccharide (LPS) released from periodontal pathogens, correlates with a poor prognosis after esophagectomy for ESCC. We therefore hypothesized that LPS influences cancer cell proliferation and disease progression in ESCC. We used 8 ESCC cell lines to investigate how LPS affects ESCC cell proliferation and migration activity. We also assessed mRNA and protein expression to determine how LPS affects cytokine production and whether blocking TLR4 signaling attenuates that effect. We also used a mouse xenograft model to investigate whether LPS upregulates ESCC tumor progression in vivo. We then determined whether C-C motif chemokine ligand 2 (CCL2) expression in clinical samples correlates with 5-year overall survival (OS) and disease-specific survival (DSS) in ESCC patients after esophagectomy. LPS significantly upregulated cell proliferation and migration in all ESCC lines. It also upregulated CCL2 production. In vivo, subcutaneous LPS administration significantly increased ESCC tumor volume in mice. In clinical samples, high CCL2 expression significantly correlated with 5-year OS and DSS. There was also a significant correlation between CCL2 and TLR4 expression status, suggesting the involvement of an LPS-TLR4-CCL2 cascade in clinical settings. LPS significantly upregulates cell proliferation and tumor progression through an LPS-TLR4-CCL2 cascade and influences prognosis after esophagectomy for ESCC. This suggests improving the oral environment has the potential to improve the prognosis of ESCC patients after esophagectomy.

KLC1-ROS1 Fusion Exerts Oncogenic Properties of Glioma Cells via Specific Activation of JAK-STAT Pathway.

Here, we investigated the detailed molecular oncogenic mechanisms of a novel receptor tyrosine kinase (RTK) fusion, KLC1-ROS1, with an adapter molecule, KLC1, and an RTK, ROS1, discovered in pediatric glioma, and we explored a novel therapeutic target for glioma that possesses oncogenic RTK fusion. When wild-type ROS1 and KLC1-ROS1 fusions were stably expressed in the human glioma cell lines A172 and U343MG, immunoblotting revealed that KLC1-ROS1 fusion specifically activated the JAK2-STAT3 pathway, a major RTK downstream signaling pathway, when compared with wild-type ROS1. Immunoprecipitation of the fractionated cell lysates revealed a more abundant association of the KLC1-ROS1 fusion with JAK2 than that observed for wild-type ROS1 in the cytosolic fraction. A mutagenesis study of the KLC1-ROS1 fusion protein demonstrated the fundamental roles of both the KLC1 and ROS1 domains in the constitutive activation of KLC1-ROS1 fusion. Additionally, in vitro assays demonstrated that KLC1-ROS1 fusion upregulated cell proliferation, invasion, and chemoresistance when compared to wild-type ROS1. Combination treatment with the chemotherapeutic agent temozolomide and an inhibitor of ROS1, JAK2, or a downstream target of STAT3, demonstrated antitumor effects against KLC1-ROS1 fusion-expressing glioma cells. Our results demonstrate that KLC1-ROS1 fusion exerts oncogenic activity through serum-independent constitutive activation, resulting in specific activation of the JAK-STAT pathway. Our data suggested that molecules other than RTKs may serve as novel therapeutic targets for RTK fusion in gliomas.

B-377 Mirsnps as Potential Colorectal Cancer Biomarkers: An Overview

Abstract Background Colorectal cancer (CRC), also known as colon cancer, is a neoplasm that affects the colon (large intestine) or rectum. CRC stands out for being the third most frequent neoplasm in the world population, being the second cancer with the highest mortality rate. Single nucleotide polymorphisms (SNPs) located in miRNAs genes, known as miRSNPs, may be associated with dysregulated expression of miRNAs in neoplasms. The objective of this review was to highlight the SNPs that alter the biogenesis or processing of miRNAs associated with CRC as potential biomarkers of this disease, according to the literature. Methods A search of original articles in the literature was performed in PubMed, Embase and Web of Science databases. The bibliographic search included scientific articles published between 2008 and 2022, with the objective of investigating associations between miRSNPs and CRC. Exclusion criteria were brief communications and other reviews on the subject. A total of 263 articles were detected in the preliminary search, of which 133 were excluded for being replicated, 34 for being reviewers, 3 for being paid and 40 for dealing with other subjects. Thus, of the remaining 53 studies, 35 were excluded after reading because they included studies that described SNPs located in miRNA genes and SNPs located in miRNA binding sites. Finally, 18 articles that included SNPs located in the regulatory regions of miRNAs were selected for this review. The list of genes generated for each of the targets was performed using the online tool miRBase and miRDB, for prediction of miRNA targets and functional annotations. The miRTarBase online database was consulted to select target miRNAs and possible interactions. By crossing the list of genes targeted by miRNAs to identify the biological pathways, those with the greatest biological relevance were chosen using the Enrichr online tool. Results This review covered 18 studies related to SNPs located in regulatory regions of mature miRNAs that may influence the occurrence and progression of CRC. In order to characterize the metabolic profile induced by each target of the miRNAs selected in this review, it was possible to observe that miR-196, miR-211, miR-30a, miR-143, miR-27 and miR-146a are more strongly associated with positive regulation of transcription. Cytokine-mediated signaling pathway and regulation of cell migration were most strongly associated with miR-143 and miR-146a. Furthermore, it was identified that the upregulation of the apoptosis process was strongly associated with miR-30a, miR-143, miR-27a and miR-146a. MiR-211, miR-143, mi-27a and miR-146a were strongly associated with the upregulation of cell proliferation. Finally, with the objective of evaluating the correlation between the selected miRNAs. It was possible to observe a greater correlation between miR-27a and miR-30a. Conclusion Studies related to miRSNPs have been of great interest in recent years in order to find promising and less invasive new biomarkers, sensitive and specific for the early detection of CRC. Our study aims to help better understand the pathways that cause the development of CRC and possible biomarkers to predict its development and consequently reduce morbidity and mortality and improve patient survival.

Ethylene response factor ERF022 is involved in regulating Arabidopsis root growth.

Ethylene response factors (ERFs) are involved in the regulation of plant development processes and stress responses. In this study, we provide evidence for the role of ERF022, a member of the ERF transcription factor group III, in regulating Arabidopsis root growth. We found that ERF022-loss-of-function mutants exhibited increased primary root length and lateral root numbers, and also morphological growth advantages compared to wild-type. Further studies showed that mutants had enhanced cell size in length in the root elongation zones. These results were accompanied by significant increase in the expression of cell elongation and cell wall expansion related genes SAUR10, GASA14, LRX2, XTH19 in mutants. Moreover, ERF022-mediated root growth was associated with the enhanced endogenous auxin and gibberellins levels. Our results suggest that loss-of-function of ERF022 up-regulated the expression of cell elongation and cell wall related genes through auxin and gibberellins signal in the regulation of root growth. Unexpectedly, ERF022 overexpression lines also showed longer primary roots and more lateral roots compared to wild-type, and had longer root apical meristematic zone with increased cell numbers. Overexpression of ERF022 significantly up-regulated cell proliferation, organ growth and auxin biosynthesis genes EXO, HB2, GALK2, LBD26, YUC5, which contribute to enhanced root growth. Altogether, our results provide genetic evidence that ERF022 plays an important role in regulating root growth in Arabidopsis thaliana.

The E3 ubiquitin ligase SCF (FBXW10)-mediated LATS2 degradation regulates angiogenesis and liver metastasis in colorectal cancer.

F-box and WD repeat domain containing 10 (FBXW10) is a member of the FBXW subgroup that contains the WD40 domain. FBXW10 has been rarely reported in colorectal cancer (CRC) and its mechanism is unclear. To investigate the role of FBXW10 in CRC, we conducted in vitro and in vivo experiments. Through the database and our clinical samples, we found that FBXW10 expression was up-regulated in CRC, and it was positively correlated with CD31 expression. CRC patients with high FBXW10 expression levels had a poor prognosis. Overexpression of FBXW10 up-regulated cell proliferation, migration and vascular formation, while knockdown of FBXW10 had the opposite effects. Studies on the mechanism of FBXW10 in CRC showed that FBXW10 could ubiquitinate large tumor suppressor kinase 2 (LATS2) and promote its degradation with the Fbox region of FBXW10 played an essential role in this process. In vivo studies demonstrated that knockout of FBXW10 inhibited tumor proliferation and reduced liver metastasis. In conclusion, our study proved that FBXW10 was significantly overexpressed in CRC and was involved in the pathogenesis of CRC by affecting angiogenesis and liver metastasis. Mechanistically, FBXW10 degraded LATS2 through ubiquitination. Therefore, FBXW10-LATS2 can be used as a therapeutic target for CRC in subsequent studies.

Suprabasin enhances the invasion, migration, and angiogenic ability of oral squamous cell carcinoma cells under hypoxic conditions.

Suprabasin (SBSN) is a secreted protein that is isolated as a novel gene expressed in differentiated keratinocytes in mice and humans. It induces various cellular processes such as proliferation, invasion, metastasis, migration, angiogenesis, apoptosis, therapy and immune resistance. The role of SBSN was investigated in oral squamous cell carcinoma (OSCC) under hypoxic conditions using the SAS, HSC‑3, and HSC‑4 cell lines. Hypoxia induced SBSN mRNA and protein expression in OSCC cells and normal human epidermal keratinocytes (NHEKs), and this was most prominent in SAS cells. The function of SBSN in SAS cells was analyzed using 3‑(4,5‑dimethylthiazol‑2‑yl)‑2,5‑diphenyltetrazolium bromide (MTT); 5‑bromo‑2'‑deoxyuridine (BrdU); cell cycle, caspase 3/7, invasion, migration, and tube formation assays; and gelatin zymography. Overexpression of SBSN decreased MTT activity, but the results of BrdU and cell cycle assays indicated upregulation of cell proliferation. Western blot analysis for cyclin‑related proteins indicated involvement of cyclin pathways. However, SBSN did not strongly suppress apoptosis and autophagy, as revealed by caspase 3/7 assay and western blotting for p62 and LC3. Additionally, SBSN increased cell invasion more under hypoxia than under normoxia, and this resulted from increased cell migration, not from matrix metalloprotease activity or epithelial‑mesenchymal transition. Furthermore, SBSN induced angiogenesis more strongly under hypoxia than under normoxia. Analysis using reverse transcription‑quantitative PCR showed that vascular endothelial growth factor (VEGF) mRNA was not altered by the knockdown or overexpression of SBSN VEGF, suggesting that VEGF is not located downstream of SBSN. These results demonstrated the importance of SBSN in the maintenance of survival and proliferation, invasion and angiogenesis of OSCC cells under hypoxia.

Effects of a Functional Ice Cream Enriched with Milk Proteins on Bone Metabolism: A Feasibility Clinical Study and In Vitro Investigation.

Background: Milk proteins (MPs) and their derivative whey proteins (WPs) are important components of human diet that might prevent bone loss. We aimed to investigate the effects of MP on the bones of postmenopausal women, along with the effects of WP on osteoblast cells. Methods: We conducted a feasibility controlled clinical study with 62 postmenopausal women who were asked to consume an MP-enriched ice cream. We also investigated the effect of WP on the ERK1/2 and AKT pathways, RUNX2, alkaline phosphatase, RANKL/OPG ratio, and COL1A of Saos-2. Results: After 12 weeks, we found a greater bone mineral density and bone alkaline phosphatase reduction in women who consumed the MP-enriched ice cream compared to the control group (p = 0.03 and p = 0.02, respectively). In Saos-2 cells, WP upregulated ERK1/2 and AKT pathways (p = 0.002 and p = 0.016), cell proliferation (p = 0.03), and osteoblast differentiation markers, along with downregulating RANKL/OPG (p < 0.001). Moreover, the inhibition of ERK1/2 by PD184253 reverted the effects on both the RUNX2 and ALP mRNA expression and cells proliferation (p = 0.028, p = 0.004, and p = 0.003, respectively) when treated with WP. Conclusions: WP upregulates cell proliferation, RUNX2, and alkaline phosphatase through the activation of the ERK1/2 pathways on Saos-2. These mechanisms probably contribute to preventing bone loss in postmenopausal women.

PHF19 Inhibits Multiple Myeloma Cell Response to Immunotherapy Via Promoting Immunosuppressive Microenvironment

Introduction: The multiple myeloma (MM) bone marrow (BM) microenvironment promotes tumor cell growth, survival, and drug resistance, while also conferring immunosuppression. This involves crosstalk between the myeloma cells and the components of BM microenvironment which can be facilitated or hindered by epigenetic modifiers altering the gene expression of receptors. Among those, regulatory T (Tregs) cells are essential to control peripheral tolerance and response to foreign and tumor antigens. Tregs within the BM niche play a significant role in the suppression of antitumor immune responses and limiting the beneficial response of immunotherapy in MM. Here, we investigated the impact of the Polycomb-like protein PHF19/PCL3, an epigenetic gene whose high level predicts poor prognosis in patients with MM, on the immunosuppressive BM microenvironment. Methods & Results: We first investigated how PHF19 overexpression would affect the microenvironment by using an ex vivo co-culture of MM cells with PBMCs or purified CD3+ T cells. MM cells that overexpressed PHF19 by lenti-virus infection (PHF19-OE cells) induced 2-4-fold higher conversion of conventional T cells (Tcon) into induced regulatory T cells (iTregs) (n=5, day 4). In addition, PHF19-OE MM cells increased the frequency of iTreg at later time points (day 7) and upregulated cell proliferation (CCND1, CCND2), anti-apoptosis (BCL2, BCL2L1), and Treg-related genes (TGFβ1, IL-10, Foxp3) (P < .0001). Moreover, enhanced checkpoint marker (PD1, TIM3, LAG3) expression was observed on Tcon co-cultured with PHF19-OE vs control MM cells. There were elevated IL-10 and TGFβ1 levels in the culture supernatants of PHF19-OE MM cells, alone and in co-cultures, and antibodies to these cytokines partially neutralized the Treg inhibition of Tcon proliferation triggered by PHF19-OE MM cells. Thus, PHF19 seems to be important in facilitating the crosstalk between MM cells and immune cells in the BM. CD38 expression on the MM cell surface is commonly targeted in immunotherapy. However, CD38 expression was downregulated on PHF19-OE vs -KD MM cells, and PHF19 mRNA expression was negatively correlated to CD38 expression in patient MM cells (Spearman's ρ = -0.38 and -0.2 for GSE5900 and GSE31161, P < .004). As expected, PHF19-OE MM cells were less susceptible than PHF19-KD MM cells to in vitro NK-mediated lysis induced by daratumumab (dara) or isatuximab, which target CD38, as evidenced by fewer CD107a+ and IFN γ+ NK effector cells. Moreover, dara-induced cytotoxicity was diminished in xenografted NSG mice with PHF19-OE vs control cells and reconstituted with human NK cells. These resistant PHF19-OE AMO1 tumors had lower expression of CD38, STAT1, and IFN-related genes, which produced higher levels of human κ light chain, soluble BCMA, and TGFβ1 in matched serum samples. Accordingly, PHF19-OE resulted in significantly decreased mouse survival. To find the mediators of the immunosuppression induced by PHF19, we used ATAC-seq and CHIP-seq analysis in MM cell lines. We found that a number of genes essential for T cell-mediated tumor immunity was downregulated in the PHF19 OE group vs control, including key genes in the interferon-γ (IFN-γ) signaling pathway, which is a pathway that leads to cytokine-mediated cell death. Among the genes of the IFN-γ pathway, 56 genes had less chromatin accessibility in PHF19 OE vs control cells. RNA-seq analysis confirmed that PHF19 OE repressed the expression of CD38 and the IFN-γ and IFN-α signaling pathway. Furthermore, PHF19 OE altered the expression of genes in NF-κB signaling triggered by TNF-α signaling pathway, which are associated with resistance to immunotherapy. Finally, activation of the PI3K/AKT pathway was also identified in PHF19 OE cells, which directly promotes MM cell proliferation and survival, as well as enhances resistance to immunotherapy via upregulation of PD-L1 on the MM cell surface. Conclusions: Taken together, these data indicate that PHF19 promotes an immunosuppressive BM milieu by promoting iTreg frequency and the expression of immunosuppressive cytokines, as well as decreasing CD38 expression on MM cells and diminishing their sensitivity to IFN-γ-mediated killing. These results suggest the potential clinical utility of targeting PHF19 to overcome immunosuppression and thereby enhance or restore sensitivity to CD38-targeted therapy in MM.

The C-terminus of Gain-of-Function Mutant p53 R273H Is Required for Association with PARP1 and Poly-ADP-Ribose.

The C-terminal region of mtp53 confers GOF activity that mediates mtp53-PARP1 and PAR interactions assisting DNA replication, thus implicating new biomarkers for PARP inhibitor therapy.

Effect of Puerarin on Osteogenic Differentiation in vitro and on New Bone Formation in vivo

PurposePuerarin (C21H20O10) is a phytoestrogen that possesses various pharmacological effect, and several researches have revealed the relationship between puerarin and bone metabolism. This study was aimed to evaluate the potential influence of puerarin on the proliferation and osteogenic differentiation of rat bone marrow-derived mesenchymal stem cells (BMSCs) as well as on new bone formation following rapid maxillary expansion (RME) model in rats.MethodsRat BMSCs were adopted, and the cell proliferation was detected by cell-counting kit-8 (CCK-8) assay in vitro experiments. Alkaline phosphatase (ALP) activity and alizarin red staining were analyzed quantitatively to show extracellular matrix mineralization. The mRNA and protein expression levels were used to detect osteogenic differentiation of BMSCs. In vivo bone regeneration was analyzed in a rat RME model. Eighteen 6-week-old male Wistar rats were divided into 3 groups: group 1 without any treatment, group 2 received RME and saline solution (15mg/kg), group 3 received RME and puerarin solution (15mg/kg). After 2 weeks, micro-computed tomography (Micro-CT), hematoxylin and eosin (HE) staining, and Masson staining were used to detect the new bone formation and morphological changes. Besides, ALP and bone morphogenetic protein 2 (BMP2) expression levels in mid-palatal suture were evaluated by immunohistochemical staining.ResultsThe results showed that puerarin upregulates cell proliferation dose-dependently. ALP activity and mineralized matrix generation were clearly enhanced at certain specific concentrations (10−5 and 10−6 mol/L); the expression levels of the osteoblast-related genes and proteins were increased. The measurement of micro-CT imaging revealed that puerarin significantly promoted new bone formation. Concomitantly, the histological examinations showed that puerarin solution enhanced osteogenesis in mid-palatal suture.ConclusionThose works indicated that puerarin regulates osteogenesis in vitro and exerts a beneficial impact on bone regeneration in vivo, revealing that puerarin treatment may become one of the potential keys for improving the stability and preventing relapse of RME.

The role of mesenchymal-epithelial transition factor (c-MET) in cancer development and treatments

The mesenchymal-epithelial transition factor (c-MET) is classified into the tyrosine kinase receptor family. Its indispensable role in regulating the cell cycle through various downstream pathways has made it one of the most essential transmembrane receptors. A MET receptor monomer contains six domains, and each has its own function when activated by hepatocyte growth factor (HGF). Due to its complexity, c-MET aberrations including point mutations, amplification, protein overexpression, splicing site mutation, fusion, and HGF autocrine or paracrine upregulate cell proliferation and are common in most aggressive cancer types such as colorectal cancer, lung cancer, liver cancer, and glioblastoma. Correspondingly, cancer therapies targeting c-MET have been researched for decades. This review presented the mechanisms under c-MET activation, discussed its role in cancer development, and summarized recent advancements in clinical trials. c-MET inhibitors, especially combined with other therapeutic inhibitors, appeared to be a promising strategy when taking selectivity, resistance, and tolerability into account.

Functional Roles and Targets of COVID-19 in Blood Cells Determined Using Bioinformatics Analyses.

Background:Coronavirus disease 2019 (COVID-19), caused by novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is a global epidemic with a high mortality rate. In this study, our goal was to identify the function and associated targets of SARS-CoV-2 from circulating monocytes in the blood and peripheral blood mononuclear cell (PBMC) dataset of patients with COVID-19.Methods:The Gene Expression Omnibus database (GSE164805 and GSE180594) was used to identify differentially expressed genes (DEGs). Gene ontology function analysis and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses of the DEGs were performed using the DAVID database.Results:Gene ontology analysis of DEG revealed that GSE164805 and GSE180594 were involved in the regulation of cell migration, upregulation of cell proliferation, and in the activation of the mitogen-activated protein kinase signaling pathway. Kyoto Encyclopedia of Genes and Genomes analysis of GSE164805 revealed that the DEGs were enriched in peroxisome, melanogenesis, and actin regulation. Peroxisome genes were highly expressed in patients with mild and severe COVID-19. Bioinformatics analysis to compare GSE180594 and public data for the single-cell atlas of the peripheral immune response in patients with COVID-19 showed that interferon-associated genes were highly increased in acute COVID-19 PBMC and in CD14+ and CD16+ monocytes from patients with COVID-19.Conclusions:We comprehensively analyzed the blood cell gene expression profile data of patients with COVID-19 using bioinformatics methods to preliminary understand the functions and associated targets of DEGs in the blood cells of these patients. Thus, our data provide targets for potential therapies against COVID-19.

Benefit of Serum Thymidine Kinase 1 Concentration for Risk Assessment from Gastric Neoplasms Progression to Carcinomas: A Systematic Review and Meta-analysis

Background: Human Thymidine kinase 1 (hTK1), a key enzyme involved in the DNA synthesis during S-phase of the cell cycle and upregulation of cell proliferation, thus it is reliable tumor proliferating biomarker for assessment of tumor proliferation rate in serum and in tissue in oncology. This meta-analysis is investigation whether the serum TK1 concentration（STK1p）based on hTK1-IgY-polyclonal-antibody can provide a benefit for risk assessment from gastric neoplasm progression to gastric carcinoma (GC) as well as for evaluation of treatment effect in GC. 1.2. Methods: Relevant studies were identified from the following electronic databases: PubMed, Embase, Chinese National Knowledge Infrastructure, Wanfang, the Chongqing VIP Chinese Science and Technology Periodical Database and Chinese Biomedical Literature Database from May 1, 2008, until November 30, 2021. Study quality was evaluated by the modified Newcastle-Ottawa Scale. Pooled estimates of weighted mean difference (MD), sensitivity analyses and publication bias were evaluated using Review manager and STATA software. RESULTS: Twenty-seven studies included to 1,909 GCs and 1,229 neoplasms (superficial gastritis, chronic gastritis, atrophic and gastric ulcers) and 2,260 tumor-frees. The results showed that STK1p levels increased significantly in the following manner (P&lt;.0.006): tumor-free &lt; superficial gastritis &lt; chronic gastritis &lt; atrophic gastritis &lt; gastric ulcer &lt; GC. The STK1p value of the GCs one-month-post-surgery decline significantly by 66% compared to pre-surgery. 1.3. Conclusion: STK1p value has the potential to be an early risk warning indicator for early gastric neoplasms progression to GCs and to be a helpful index for monitoring the response to surgery. 1.4. Core Tip: In spite of improvements in diagnosis and treatment the long-term survival rate for a large number of GCs is still dismal. Hence, it is important to explore serum biomarkers for early prediction of risk gastric neoplasms into GCs. In this meta-analysis, the STK1p as a prognostic biomarker was investigated showed significantly higher in GCs compere to different neoplasm and tumor-free people and was significantly reduced one-month post-surgery. It demonstrates that STK1p as tumor proliferating serum-biomarker, can be not only used for early risk gastric neoplasm progression and evaluation of treatment effect in GC patients, but also for a variety of other tumor types in clinical setting.

Prognostic comparative genes predict targets for sorafenib combination therapies in hepatocellular carcinoma

With the increasing incidence and mortality of human hepatocellular carcinoma (HCC) worldwide, revealing innovative targets to improve therapeutic strategies is crucial for prolonging the lives of patients. To identify innovative targets, we conducted a comprehensive comparative transcriptome analysis of 5,410 human HCCs and 974 mouse liver cancers to identify concordantly expressed genes associated with patient survival. Among the 664 identified prognostic comparative HCC (pcHCC) genes, upregulated pcHCC genes were associated with prognostic clinical features, including large tumor size, vascular invasion and late HCC stages. Interestingly, after validating HCC patient prognoses in multiple independent datasets, we matched the 664 aberrant pcHCC genes with the sorafenib-altered genes in TCGA_LIHC patients and found these 664 pcHCC genes were enriched in sorafenib-related functions, such as downregulated xenobiotic and lipid metabolism and upregulated cell proliferation. Therapeutic agents targeting aberrant pcHCC genes presented divergent molecular mechanisms, including suppression of sorafenib-unrelated oncogenic pathways, induction of sorafenib-unrelated ferroptosis, and modulation of sorafenib transportation and metabolism, to potentiate sorafenib therapeutic effects in HCC combination therapy. Moreover, the pcHCC genes NCAPG and CENPW, which have not been targeted in combination with sorafenib treatment, were knocked down and combined with sorafenib treatment, which reduced HCC cell viability based on disruption to the p38/STAT3 axis, thereby hypersensitizing HCC cells. Together, our results provide important resources and reveal that 664 pcHCC genes represent innovative targets suitable for developing therapeutic strategies in combination with sorafenib based on the divergent synergistic mechanisms for HCC tumor suppression.

Substance P and Neurokinin 1 Receptor in Chronic Inflammation and Cancer of the Head and Neck: A Review of the Literature.

Head and neck cancer is a growing worldwide public health problem, accounting for approximately 1,500,000 new cases and 500,000 deaths annually. Substance P (SP) is a peptide of the tachykinin family, which has roles related to a large number of physiological mechanisms in humans. The implications of SP in carcinogenesis have recently been reported through the stimulation of the neurokinin 1 receptor (NK1R), or directly, through the effects derived from the constitutive activation of NK1R. Consequently, SP/NK1R seems to play relevant roles in cancer, upregulating cell proliferation, cell migration and chronic inflammation, among other oncogenic actions. Furthermore, there is growing evidence pointing to a central role for SP in tumour progression, singularly so in laryngeal and oral squamous cell carcinomas. The current narrative review of the literature focuses on the relationship between the SP/NK1R system and chronic inflammation and cancer in the head-and-neck region. We described a role for SP/NK1R in the transition from chronic inflammation of the head and neck mucosa, to preneoplastic and neoplastic transformation and progression.

Valsartan- and melatonin-supported adipose-derived mesenchymal stem cells preserve renal function in chronic kidney disease rat through upregulation of prion protein participated in promoting PI3K-Akt-mTOR signaling and cell proliferation

This study tested the hypothesis that valsartan (Val) and melatonin (Mel)-assisted adipose-derived mesenchymal stem cells (ADMSCs) preserved the residual renal function in chronic kidney disease (CKD) rat through promoting cellular-prior-protein (PrPC) to upregulate PI3K/Akt/mTOR signaling and cell proliferation. In vitro study demonstrated that as compared with CKD-derived-ADMSCs, Val/Mel/overexpression of PrPC-treated CKD derived-ADMSCs significantly upregulated cell proliferation and protein expressions of PrPC and phosphorylated (p)-PI3K/p-Akt/p-mTOR, and downregulated oxidative stress (all p < 0.001). Rats (n = 42) were categorized into group 1 (sham-operated-control), group 2 (CKD), group 3 (CKD + ADMSCs/1.2 ×106 cells) + Mel/20 mg/kg/day), group 4 (CKD + siRNA-PrPC-ADMSCs/1.2 ×106 cells), group 5 (CKD + ADMSCs/1.2 ×106 cells + Val/20 mg/kg/day) and group 6 (CKD + Val + Mel). By day 35, the kidney specimens were harvested and the result showed that the protein expression of PrPC was highest in group 1, lowest in groups 2/4 and significantly lower in group 6 than in groups 3/5, but it was similar in groups 3/5 (all p < 0.0001). The protein expressions of cell-stress-signaling (p-PI3K/p-Akt/p-mTOR) and cell-cycle activity (cyclin-D1/clyclin-E2/Cdk2/Cdk4) exhibited an identical pattern, whereas the protein expressions of oxidative-stress (NOX-1/NOX-2)/mitochondrial fission (PINK1/DRP1)/apoptosis (cleaved-capsase3/cleaved-PARP) and fibrosis (TFG-ß/Smad3) as well as creatinine/BUN levels, ratio of urine-protein to urine-creatine and kidney-injured score exhibited an opposite pattern of PrPC among the groups (all p < 0.0001). In conclusion, Mel/Val facilitated-ADMSCs preserved renal architecture and function in CKD rat through promoting PrPC to regulate the cell proliferation/oxidative-stress/cell-stress signalings.

Characterization of a Mouse Model of Inducible Frailty: The Humanized IL-6 Mouse

The cytokine interleukin-6 (IL-6) has pleiotropic effects in aging and is elevated in frail older adults. We have developed a conditional mouse model to better characterize the role of IL-6 in promoting frailty and age-related mitochondrial dysregulation. The human IL-6 (hIL-6) knock-in mouse (TetO-hIL6) was developed utilizing CRISPR/Cas9 technology with transgene donor vector containing a tetracycline response element promoter driving expression of hIL-6 cDNA. Male TetO-hIL6 mice were treated with doxycycline-containing water for six weeks starting at 8 months old. RNAseq analysis of whole blood demonstrated significant upregulation of pro-inflammatory related markers at 6 weeks compared to baseline and upregulated cell proliferation and metabolism pathways. Physical testing of TetO-hIL6 mice before and after hIL-6 induction demonstrated decreased grip strength (p =0.003), decreased running capacity (p = 0.02), and 40% increase in falls off of the treadmill (p = 0.001). Induced mice also demonstrated decreased basal body temperature (p < 0.001). Given the significant dysregulation of metabolism-related genes in RNAseq analysis and changes in basal body temperature following hIL-6 induction, we next performed untargeted metabolomics on plasma from mice at baseline and 6 weeks post-induction to better evaluate metabolic changes associated with hIL-6 elevation. We found changes in key serum metabolites, including circulating adenosine triphosphate (56% reduction, p = 0.02), pyruvate (35% reduction, p = 0.0006), alpha-ketoglutarate (47% reduction, p = 0.04), and succinate (306% increase, p = 0.001). The TetO-hIL6 mouse model allows for induction of hIL-6 at various timepoints across the lifespan and demonstrates features of a frailty phenotype.