Targeting Src Research Articles

Mechanism of anti-hyperplasia of mammary glands of Xihuang Pills blood-entering component based on UPLC-Q-TOF-MS and network pharmacology

In this study, based on network pharmacology and molecular docking method, the mechanism of anti-hyperplasia of mammary glands of Xihuang Pills blood-entering components was explored, and the efficacy and key targets of Xihuang Pills blood-entering components were experimentally verified by MCF-10A proliferation model of human mammary epithelial cells. In order to clarify the material basis and mechanism of Xihuang Pills in realizing anti-hyperplasia of mammary glands, the blood-entering components of Xihuang Pills were qualitatively analyzed by UPLC-Q-TOF-MS, and 22 blood-entering components were identified. By taking the blood-entering components as the research object, the network pharmacology prediction and molecular docking verification were carried out, and finally, three key targets were screened out, namely JAK1, SRC, and CDK1. In vitro experiments show that Xihuang Pills can inhibit the proliferation of MCF-10A cells, promote the apoptosis of MCF-10A cells, and reduce the expression of JAK1, SRC, and CDK1 targets in cells. To sum up, Xihuang Pills can promote the apoptosis of mammary epithelial cells by regulating the expression of JAK1, SRC, and CDK1 and then play an anti-hyperplasia role, which provides an experimental basis for clarifying the material basis of Xihuang Pills for anti-hyperplasia effect.

Overcoming BCR::ABL1 dependent and independent survival mechanisms in chronic myeloid leukaemia using a multi-kinase targeting approach

BackgroundDespite improved patient outcome using tyrosine kinase inhibitors (TKIs), chronic myeloid leukaemia (CML) patients require life-long treatment due to leukaemic stem cell (LSC) persistence. LSCs reside in the bone marrow (BM) niche, which they modify to their advantage. The BM provides oncogene-independent signals to aid LSC cell survival and quiescence. The bone-morphogenetic pathway (BMP) is one pathway identified to be highly deregulated in CML, with high levels of BMP ligands detected in the BM, accompanied by CML stem and progenitor cells overexpressing BMP type 1 receptors- activin-like kinases (ALKs), especially in TKI resistant patients. Saracatinib (SC), a SRC/ABL1 dual inhibitor, inhibits the growth of CML cells resistant to the TKI imatinib (IM). Recent studies indicate that SC is also a potent ALK inhibitor and BMP antagonist. Here we investigate the efficacy of SC in overcoming CML BCR::ABL1 dependent and independent signals mediated by the BM niche both in 2D and 3D culture.MethodsCML cells (K562 cell line and CML CD34+ primary cells) were treated with single or combination treatments of: IM, SC and the BMP receptors inhibitor dorsomorphin (DOR), with or without BMP4 stimulation in 2D (suspension) and 3D co-culture on HS5 stroma cell line and mesenchymal stem cells in AggreWell and microfluidic devices. Flow cytometry was performed to investigate apoptosis, cell cycle progression and proliferation, alongside colony assays following treatment. Proteins changes were validated by immunoblotting and transcriptional changes by Fluidigm multiplex qPCR.ResultsBy targeting the BMP pathway, using specific inhibitors against ALKs in combination with SRC and ABL TKIs, we show an increase in apoptosis, altered cell cycle regulation, fewer cell divisions, and reduced numbers of CD34+ cells. Impairment of long-term proliferation and differentiation potential after combinatorial treatment also occurred.ConclusionBMP signalling pathway is important for CML cell survival. Targeting SRC, ABL and ALK kinases is more effective than ABL inhibition alone, the combination efficacy importantly being demonstrated in both 2D and 3D cell cultures highlighting the need for combinatorial therapies in contrast to standard of care single agents. Our study provides justification to target multiple kinases in CML to combat LSC persistence.

Integrating metabolic profile and network pharmacology to explore the chemical essence of Radix Ginseng Rubra for attenuating heart failure

Radix Ginseng Rubra (RGR), a well-known edible-medicinal herb, has been widely used to attenuate heart failure (HF). However, the active ingredients and mechanism of RGR for attenuating HF have not been illuminated. Thus, the metabolic profile in vivo of RGR was investigated by ultra-performance liquid chromatography quadrupole time-of-flight tandem mass spectrometry and further the compounds identified in serum and heart were employed to search for the potential active compounds and targets via network pharmacology and molecular docking. 41 chemical compounds were identified in RGR aqueous extract. 10 prototypes and 8 metabolites were identified or tentatively characterized in vivo. Additionally, two compounds with 51 corresponding targets had been speculated to improve HF, and the role of ginsenoside Rh4 (P8) in RGR for attenuating HF by interfering with MAP2K1 and SRC targets was verified by molecular docking. Therefore, the potential active ingredients and targets were expected to be the basis of RGR therapy for HF.

Artesunate Sensitizes human hepatocellular carcinoma to sorafenib via exacerbating AFAP1L2-SRC-FUNDC1 axis-dependent mitophagy

ABSTRACT Sorafenib is the most widely used first-line drug for the treatment of the advanced hepatocellular carcinoma (HCC). Unfortunately, sorafenib resistance often limits its therapeutic efficacy. To evaluate the efficacy of artesunate against sorafenib-resistant HCC and to investigate its underlying pharmacological mechanisms, a “sorafenib resistance related gene-ART candidate target” interaction network was constructed, and a signaling axis consisting with artesunate candidate target AFAP1L2 and sorafenib target SRC, and the downstream FUNDC1-dependent mitophagy was identified as a major contributor to the sorafenib resistance and a potential way of artesunate to mitigate resistance. Notably, our clinical data demonstrated that AFAP1L2 expression in HCC tissues was markedly higher than that in adjacent non-cancerous liver tissues (P < 0.05), and high AFAP1L2 expression was also significantly associated with an unfavorable overall survival of HCC patients (P < 0.05). Experimentally, AFAP1L2 was overexpressed in sorafenib resistant cells, leading to the activation of downstream SRC-FUNDC1 signaling axis, further blocking the FUNDC1 recruitment of LC3B to mitochondria and inhibiting the activation of mitophagy, based on both in vitro and in vivo systems. Moreover, artesunate significantly enhanced the inhibitory effects of sorafenib on resistant cells and tumors by inducing excessive mitophagy. Mechanically, artesunate reduced the expression of AFAP1L2 protein, suppressed the phosphorylation levels of SRC and FUNDC1 proteins, promoted the FUNDC1 recruitment of massive LC3B to mitochondria, and further overactivated the mitophagy and subsequent cell apoptosis of sorafenib resistant cells. In conclusion, artesunate may be a promising strategy to mitigate sorafenib resistance in HCC via exacerbating AFAP1L2-SRC-FUNDC1 axis-dependent mitophagy. Abbreviations: AFAP1L2, actin filament associated protein 1 like 2; ANOVA, analysis of variance; ANXA5, annexin V; ART: artesunate; CETSA, cellular thermal shift assay; CI: combination index; CO-IP: co-immunoprecipitation; CQ: chloroquine; CT, computed tomography; [18F]-FDG, fluoro-2-D-deoxyglucose F18; FUNDC1: FUN14 domain containing 1; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; HCC, hepatocellular carcinoma; H&E Staining: hematoxylin – eosin staining; HepG2R, sorafenib resistant HepG2; IF, immunofluorescence; IHC, immunohistochemistry; LAMP1: lysosomal associated membrane protein 1; MAP1LC3B/LC3B: microtubule associated protein 1 light chain 3 beta; miR, microRNA; mRNA: messenger RNA; OE, overexpression; OS, overall survival; PET, positron emission tomography; qRT-PCR: quantitative real-time PCR; sh, short hairpin; shNC: negative control shRNA; shAFAP1L2: short hairpin AFAP1L2; SORA, sorafenib; SPR, surface plasmon resonance; SRC, SRC proto-oncogene, non-receptor tyrosine kinase; SUV, standardized uptake value; TEM, transmission electron microscopy; TOMM20: translocase of outer mitochondrial membrane 20.

Network pharmacology and Molecular docking-based activity of Hemidesmus indicus (L.) R.Br. in Acute myeloid leukemia : A Computational Study

ABSTRACT &#x0D; Background &#x0D; Acute myeloid leukemia (AML) is malignancy of the stem cell precursors of the myeloid lineage occurs due to variations in genetics. Incidence rate of childhood AML in Asian population is 8.4 per million. There is no exact description of AML is Ayurveda, it can be considered into Raktapitta (~bleeding disorder) disease. Hemidesmus indicus (L.) R.Br. (~H. indicus) is described for treatment of Raktapitta. The main treatment for AML is chemotherapy, and patient are searching for Ayurvedic medicines. Hence attempt is made for evaluating activity of H. indicus in AML.&#x0D; Objective&#x0D; To establish link for therapeutic activity of H. indicus in AML using Network pharmacology and molecular docking study. &#x0D; Materials and methods&#x0D; Active compound from root of H. indicus was retrieved from phytochemical based IMPPAT database. ADME (absorption, distribution, metabolism and excretion) study of retrieved active compound done with SwissADME database, and ADME qualified active compound target were obtained with having probability &gt;0.7 from SwissTargetPrediction database. Target of AML retrieved from GeneCard database having relevancy score ≥5.0. To take the common, target of active compound and AML targets from GeneCard are imported into the Venny2.1 database, and the resulting targets used for the analysis. Cytoscape3.9.1 software was used to construct the "drug-active components-target" network diagram from common targets. The PPI (protein-protein interaction) network between proteins was constructed by STRING and result exported to Cytoscape3.9.1 for network analysis to get subnetwork with key target of subnetwork and core targets of overall PPI. GO (gene ontology) and KEGG (Kyoto Encyclopedia of Genes and Genomes) pathway analysis of key target from subnetwork done with g-profiler database. Core targets were docked with their corresponding active compound to get docking score.&#x0D; Results&#x0D; Total of 66 active compound were obtained from H. indicus, On ADME screening 49 active compounds qualified. ADME qualified active compound screened for target and 149 target obtained, after removal of duplicates 81 target were remained. 806 targets of AML were screened from GeneCard database. 19 common targets were obtained between target of active compound and target of AML from GeneCard. These screened 19 targets imported in STRING database to construct PPI network, and obtained result imported into Cytoscape3.9.1 for network analysis, on analysis 1 sub networks with 11 key targets were obtained. Top five H. indicus core targets for AML obtained using “CytoHubba” plug-in. GO and KEGG enrichment analyses were performed on the above-mentioned 11 key targets from sub network, 44 MF (Molecular function), 166 BP (Biological process), 12 CC (Cellular component) and 55 pathways were obtained from the KEGG pathway analyses. Molecular docking results showed that the active component quercetin could spontaneously bind to the core targets EGFR, SRC, AKT1, KDR and IGF1R. EGFR has the best combination with quercetin.&#x0D; Conclusion &#x0D; All core targets identified through network analysis of PPI network were linked to common active compound quercetin, and on molecular docking study all core targets showed good docking score to quercetin. Hence, based on this study conclusion can be drawn that the activity of H. indicus is AML might be due to presence of quercetin active compound in it. This study generated link for usefulness of H. indicus is AML.

PP2 alleviates the progression of osteoarthritis by inhibiting Wnt/β-catenin and activating TGF-β/Smad signaling

ObjectiveWe aimed to explore the effect and mechanism of the Src inhibitor PP2 on osteoarthritis (OA) progression. MethodsThe protein expressions of Src, p-Src (y418) and p-FAK in normal and OA human chondrocytes were detected by immunofluorescence (IF) analysis. Chondrocytes from the femur and tibial plateau of 3-day-old mice were extracted and inoculated into 6-well plates. The chondrocytes were co-cultured with IL-1β and different doses of PP2, and then the degeneration of extracellular matrix was analyzed. A mouse OA model was induced by destabilizing medial meniscectomy of the right knee. Two weeks after the operation, different doses of PP2 were injected intraperitoneally. The drug was given three times a week for 6 weeks, and then the mice were sacrificed. Histopathological, IF and immunoblotting analyses were used to detect key OA catabolic markers and protein expression and related signaling. ResultsThe levels of Src, p-Src (y418) and p-FAK in the knee cartilage tissue of patients with OA were abnormally increased. After chondrocytes were co-treated with IL-1β and different doses of PP2, the results showed that PP2 reduced the abnormal increase of β-catenin, p-β-catenin and other proteins induced by IL-1β, and reversed the decrease of p-Smad3, aggrecan and collagen Ⅱ protein levels. Meanwhile, intraperitoneal injection of PP2 in vivo significantly reduced the degeneration of articular cartilage in the OA mouse model. ConclusionOur data indicate that targeting Src with PP2 protected against cartilage destruction in an OA mouse model by inhibiting Wnt/β-catenin and activating TGF-β/Smad signaling, suggesting that Src may be a potential therapeutic target for OA treatment.

Using network pharmacological analysis and molecular docking to investigate the mechanism of action of quercetin's suppression of oral cancer.

This investigation seeks to explore the mechanism of quercetin in oral cancer by incorporating network pharmacology analysis and molecular docking. First, we use the network pharmacology analysis to discover possible core targets for quercetin and oral cancer. We subsequently utilized the docking of molecules techniques to calculate the affinities of critical targets and quercetin for verification. TCMSP and the Swiss Target Prediction database found 190 quercetin action targets, while GeneCards, OMIM, PharmGkb, and the Therapeutic Target Database found 8971 oral cancer-related targets. Venny 2.1.0 online software conducted an intersection analysis of quercetin-related target information with information about oral cancer, and 172 putative quercetin-anti-oral cancer targets were examined. Six prospective core targets for quercetin treatment of oral cancer were identified from the PPI network topology analysis of 172 putative therapeutic targets. These targets include AKT1, PIK3R1, MYC, HIF1A, SRC, and HSP90AA1. GO enrichment function analysis showed that 2372 biological processes, 98 cell components, and 201 molecular functions were involved. Through enrichment analysis of the KEGG pathway, 172 signal pathways were obtained. A few examples are PI3K-AKT, HIF-1, IL-17, and other signaling pathways. The molecular docking scores of quercetin and the primary therapeutic targets AKT1, HIF1A, HSP90AA1, MYC, PIK3R1, and SRC are all less than -0.7 points, demonstrating good compatibility between the medicine and small molecules and suggesting that quercetin may affect oral cancer through the primary target. This study explores quercetin's mechanism and possible targets for oral cancer treatment, offering novel approaches. Quercetin may be a multitarget medication against oral cancer in the future.

Combined Metabolite Analysis and Network Pharmacology to Elucidate the Mechanisms of Therapeutic Effect of Melastoma dodecandrum Ellagitannins on Abnormal Uterine Bleeding.

The abnormal uterine bleeding (AUB) is complex and usually leads to severe anemia. Melastomadodecandrum (MD) is clinically used for the treatment of metrorrhagia bleeding. The MD ellagitannins (MD-ETs) had been evidenced being effective at hemorrhage, and exerts biological activities upon their metabolites including ellagic acid and urolithins. In this study, the blood-permeated metabolites from theMD-ETs were analyzed using LC-MS approach, and 19 metabolites including ellagic acid and urolithin A derivatives were identified. Furthermore, a network pharmacology analysis including the target prediction analysis, AUB target analysis, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis were conducted to reveal the relationships between "metabolites-targets-pathways", which was further verified by molecular docking analysis. The results showed that methyl ellagic acid, urolithin A and isourolithin A produced from MD-ETs can be absorbed into the blood, and might act on the core targets of VEGFA, SRC, MTOR, EGFR and CCND1. And the hemostatic effects were exerted through PI3K-Akt, endocrine resistance and Rap 1 signaling pathways. These results implied the potential effective constituents and action mechanism of MD-ETs in the therapy of AUB, which will promote the application of MD-ETs as natural agent for the treatment of gynecological bleeding diseases.

Investigating the potential anti-inflammatory mechanism of benzophenone compounds from the leaves of Aquilaria sinensis (Lour.) Gilg based on network pharmacology and molecular docking strategies

Background Aquilaria sinensis (Lour.) Gilg (ASG) has been used as traditional medicine for centuries. However, the active ingredients from leaves and their anti-inflammatory mechanism are rarely reported. The network pharmacology and molecular docking strategies were applied to explore the potential mechanisms of Benzophenone compounds from the leaves of ASG (BLASG) against inflammation. Methods BLASG-related targets were obtained from the SwissTargetPrediction and PharmMapper databases. Inflammation-associated targets were retrieved from GeneGards, DisGeNET, and CTD databases. Cytoscape software was used to draw a network diagram of BLASG and its corresponding targets. DAVID database was applied for enrichment analyses. A protein-protein interaction (PPI) network was constructed to identify the hub targets of BLASG. Molecular docking analyses were performed by AutoDockTools 1.5.6. Moreover, we used ELISA and qRT-PCR assays to validate the anti-inflammatory effects of BLASG by cell experiments. Results Four BLASG were extracted from ASG, and corresponding 225 potential targets were identified. PPI network analysis indicated that SRC, PIK3R1, AKT1, and other targets were the core therapeutic targets. Enrichment analyses revealed that the effects of BLASG are regulated by targets associated with apoptosis and inflammation-related pathways. In addition, molecular docking revealed that BLASG combined well with PI3K and AKT1. Furthermore, BLASG significantly decreased the inflammatory cytokines levels and down-regulated PIK3R1 and AKT1 gene expression in RAW264.7 cells. Conclusion Our study predicted the potential targets and pathways of BLASG against inflammation, which offered a promising strategy to reveal the therapeutic mechanism of natural active components in the treatment of diseases.

Gut microbiota and urine metabolomics based exploration of Coreopsis tinctoria Nutt. tea on type 2 diabetic mellitus

Scented tea, as an integral part of tea culture for several centuries, is important to prevent chronic metabolic diseases. Coreopsis tinctoria Nutt. (CT) tea is often used to prevent and treat diabetes, but the mechanism of action needs further study. Liquid chromatography-mass spectrometry was used to analyze 23 chemical components of CT extracts, and a “compounds-target-healthy function” network was constructed while the pathway enrichment analysis was carried out. Eleven CT compounds, including flavanomarein, luteolin, isookanin, marein, 7,3′,5′-trihydroxy-flavanone, okanin, caffeic acid, cynaroside, acetylcoreopsin, quercetin-7-O-β-D-glucoside, protocatechuic acid, were obtained through network pharmacology screening. The corresponding targets SRC, TP53, HSP90AA1, STAT3, MAPK1, PIK3R1, AKT1 and PIK3CA were associated with anti-glycemic effects. The rat model of type 2 diabetic mellitus (T2DM) was induced by a high-fat diet combined with low-dose streptozotocin injection and biochemical indexes (blood glucose, serum total cholesterol, and triglyceride) were determined. The low-dose water extract of CT can significantly regulate the total cholesterol level of diabetic rats (p < 0.01). 16S rDNA sequencing showed that CT could reverse the imbalance of intestinal flora, especially Lactobacillus, Prevotella, Succinivibrio, unidentified_Ruminococcaceae, and Treponema. Urine metabolomics observed 31 differential metabolites, mainly related to Glycerophospholipid metabolism. The correlation study showed that CT could treat T2DM by modulating the interaction of gut microbiota composition, changes in key bacterial genera, and the levels of endogenous metabolites to act on carbohydrate metabolism, amino acid metabolism, and lipid metabolism pathways. Therefore, CT water extract can effectively alleviate the changes of T2DM may be related to the combined changes of intestinal flora and metabolic characteristics.

A phase I trial of palbociclib (palbo) and bosutinib (bos) with fulvestrant (fulv) in patients (pts) with hormone receptor–positive, HER2-negative (HR+/HER2-) metastatic breast cancer (MBC) refractory to an aromatase inhibitor (AI) and a CDK4/6 inhibitor (CDK4/6i).

1079 Background: Standard therapy for HR+/HER2- MBC includes a CDK 4/6i with endocrine therapy (ET). Eventual resistance results in progression of disease (PD). A potential mechanism of CDK4/6 resistance is through Src activation. Targeting Src and ET suppresses proliferation of ER+ hormone independent breast cancer cells. Therefore, adding bos, a Src inhibitor, to palbo + fulv may overcome this resistance and restore sensitivity to CDK4/6i. Methods: This is a single arm, 3+3 dose escalation phase I trial (NCT 03854903) of bos + palbo + fulv for pts with HR+/HER2- MBC who have PD after ≥ 1 AI, ≥ 1 CDK4/6i, and &lt; 3 lines of chemo for MBC. Pts were enrolled in two cohorts, A and B, with two dose levels in cohort B. Fulv was given at standard dose. Cohort A: bos 300mg (D1-5/wk), palbo 75mg D1-21 q28 days, fulv; cohort B1: bos 300mg (D1-5/wk), palbo 100 mg D1-21 q28 days, fulv; cohort B2: bos 500mg (D1-5/wk), palbo 100 mg D1-21 q28 days, fulv. Primary objective was safety and tolerability, including MTD and recommended phase 2 dose (RP2D). Secondary objectives included ORR, and CBR 24 wks. Serum, plasma, and peripheral blood mononuclear cells were collected for Src activation, cell cycle profile arrays, and cytokine array screens. Results: 18 evaluable pts were enrolled between 4/2019 – 11/2022. To date, 16pts have completed and 2 pts remain on treatment. All pts had received prior palbo and 15/18 (83%) had received prior fulv. Median number of prior therapies in the metastatic setting was 3.2. The combination in cohort A and B1 was well tolerated. The combination in cohort B2 (bos 500mg D1-D5/wk) was toxic which resulted in bos dose reduction in all pts in the B2 cohort. There were no DLTs. The RP2D was bos 300 mg D1–5/wk, palbo 100 mg D1-21 q28 days, and standard dose fulv. No pts came off treatment due to adverse events (AEs). Most common AEs were G1/2 (9 pts) and G3/4 (7 pts) neutropenia (no neutropenic fever); G1 nausea (8 pts); G1/2 (12pts) and G3 (1pt) diarrhea; G1/2 rash (4 pts), and no G5 events. Median PFS was 5.9 months with 95% CI (3, 10.1). CBR at 6 months was 50% (9/18) (95% CI (0.26, 0.74)). The ORR was 6% (0 CR, 1 PR cohort B2), but best response was SD in 11 pts. Correlative studies underway include assessment pre and post treatment of 1) Src levels and activity to confirm that bos is acting on target and 2) RNA sequencing of Src activation and cell cycle profiles and 3) cytokine array screens to test if Bos affects inflammatory markers. Conclusions: Bos + palbo + fulv was well tolerated at the RP2D. A robust CBR of 50% was seen despite all patients having prior PD on palbo and 83% on fulv. This is an effective combination for HR+/HER2- MBC which warrants further investigation and may represent a mechanism to overcome resistance to CDK4/6i and provide another oral treatment option. Clinical trial information: NCT03854903 .

Paeonol attenuates Substance P-induced urticaria by inhibiting Src kinase phosphorylation in mast cells

BackgroundTreatment of chronic urticaria is challenging, the discovery of effective therapeutic drugs is urgently in demand. PurposeTo study the effect and mechanism of Paeonol targeting mast cells and its therapeutic effect on chronic urticaria.Study design: We developed a chronic urticaria model in vivo and mast cell model in vitro examined the effect of Paeonol in the treatment of chronic urticaria and its mechanism of action in mast cells. MethodThe anti-anaphylactoid effect of Paeonol was evaluated in PCA and systemic anaphylaxis models. The treatment role of Paeonol was studied in urticaria model. The release of cytokines and chemokines was measured using enzyme immunoassay kits. Western blot analysis was conducted to investigate phosphorylation of Src, PI3K, and PLC. In vitro kinase assays were conducted to investigate the kinase activity of Lyn, PLC, PI3K and Src. ResultsIn our study, Paeonol was able to attenuate evans blue leakage, serum histamine and chemokine release in a passive skin allergic reaction model. Simultaneously, Paeonol inhibited vasodilation and mast cell degranulation in C57BL/6 mice. Further research found that Paeonol alleviated symptoms such as erythema and rash in the Substance P-induced urticaria model, this is accompanied by inhibiting the release of related inflammatory factors. Validation experiments on mast cells in vitro found that Paeonol inhibited the activation of Src-PI3K/Lyn-PLC-NF-κB signaling pathway by crosslinking with Src kinase. Moreover, calcium influx, mast cell degranulation, cytokines generation and chemotaxis were reduced in LAD2 cells. Molecular docking experiments revealed that Paeonol is a specific antagonist targeting Src kinase in the treatment of skin diseases such as urticaria. ConclusionPaeonol, a herb-derived phenolic compound, can provide drug candidate for developing new drug in treatment of skin disease such as urticaria. Significance statementIn this study, we primarily examined the effect of Paeonol in the treatment of chronic urticaria and its mechanism of action in mast cells. Interestingly, Paeonol was found to regulate Src kinase activity downstream of MRGPRX2 triggered signaling cascade in mast cells. Therefore, this plant-derived phenolic compound may provide a therapeutic option for the treatment of chronic urticaria.

Exploring the effect of Yinzhihuang granules on alcoholic liver disease based on pharmacodynamics, network pharmacology and molecular docking

BackgroundYinzhihuang granules (YZHG) is a commonly used Chinese patent medicine for the treatment of liver disease. However, the mechanism of YZHG in alcoholic liver disease (ALD) is still unclear.MethodsThis study combined liquid chromatography-mass spectrometry technology, pharmacodynamics, network pharmacology and molecular docking methods to evaluate the potential mechanism of YZHG in the treatment of ALD.ResultsA total of 25 compounds including 4-hydroxyacetophenone, scoparone, geniposide, quercetin, baicalin, baicalein, chlorogenic acid and caffeic acid in YZHG were identified by ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS). The pharmacodynamic investigations indicated that YZHG could improve liver function and the degree of liver tissue lesions, and reduce liver inflammation and oxidative stress in ALD mice. Network pharmacology analysis showed that YZHG treated ALD mainly by regulating inflammation-related signaling pathways such as the PI3K-Akt signaling pathway. The results of the PPI network and molecular docking showed that the targets of SRC, HSP90AA1, STAT3, EGFR and AKT1 could be the key targets of YZHG in the treatment of ALD.ConclusionThis study explored the potential compounds, potential targets and signaling pathways of YZHG in the treatment of ALD, which is helpful to clarify the efficacy and mechanism of YZHG and provide new insights for the clinical application of YZHG.

MiR-654-3p targets SRC to suppress tumor growth in non-small cell lung cancer.

Around the world, cancer-related death is primarily caused by lung cancer all the time. MiR-654-3p plays an outstanding role in the development of cancer, but the mechanism of miR-654-3p in non-small cell lung cancer (NSCLC) is uncertain. For this purpose, a quantitative real-time polymerase chain reaction(qRT-PCR) was carried out to detect the expression of miR-654-3p and SRC mRNA. Western blot was used to estimate the level of SRC protein. The mimics enhanced miR-654-3p, while inhibitors knocked it down. Functional experiments were performed to evaluate the proliferation and migration capacities of cells. Flow cytometry assay was utilized to measure apoptosis rates and cell cycles of cells. TargetScan bioinformatics database was queried to identify the probable target gene for miR-654-3p. Dual-fluorescence assay was implemented to verify whether miR-654-3p targets SRC. Subcutaneous tumorigenesis was used to estimate the function of miR-654-3p in vivo. Results showed that low expression of miR-654-3p was found in NSCLC tissues and cells. Up-regulated miR-654-3p suppressed cell proliferation and migration, promoted apoptosis, and blocked cells in the G1 phase, while down-regulated miR-654-3p created the opposite results. Dual-fluorescence assay confirmed that miR-654-3p was directly bound to SRC. Compared with the control group, the effects of miR-654-3p were neutralized in the group, which was co-transfected with miR-654-3p mimics and SRC over-expression plasmids. In vivo, the tumor volume in the LV-miR-654-3p group was smaller than that in the control group. It was concluded that miR-654-3p acts in an anti-cancer role and suppresses tumor progression via regulating SRC, which lays a theoretical foundation for targeted therapy of NSCLC. MiR-654-3p is expected to be a new miRNA-based therapeutic target.

THY1 (CD90) Maintains the Adherens Junctions in Nasopharyngeal Carcinoma via Inhibition of SRC Activation

We had previously shown that THY1 (CD90) is a tumor suppressor in nasopharyngeal carcinoma (NPC) and that its down-regulation and loss of expression are associated with tumor metastasis, yet the mechanism leading to such effects remains unknown. In this study we show that tumor invasion could be suppressed by THY1 via adherens junction formation in a few NPC cell lines, and knockdown of THY1 would disrupt this cell-cell adhesion phenotype. Mechanistically, the activity of the SRC family kinase (SFK) member, SRC, and canonical Wnt signaling were dramatically reduced when THY1 was constitutively expressed. Previous studies by others have found that high levels of SRC activity in NPCs are associated with EMT and a poor prognosis. We hypothesized that THY1 can suppress tumor invasion in NPC via inhibition of SRC. By gene silencing of SRC, we found that the in vitro NPC cell invasion was significantly reduced and adherens junctions were restored. Through proteomic analysis, we identified that platelet-derived growth factor receptor β (PDGF-Rβ) and protein tyrosine phosphatase nonreceptor type 22 (PTPN22) are novel and potential binding partners of THY1, which were subsequently verified by co-immunoprecipitation (co-IP) analysis. The ligand of PDGF-Rβ (PDGF-BB) could highly induce SRC activation and NPC cell invasion, which could be almost completely suppressed by THY1 expression. On the other hand, the PTPN22 siRNA could enhance both the SRC activities and the cell invasion and could also disrupt the adherens junctions in the THY1-expressing NPC cells; the original THY1-induced phenotypes were reverted when the PTPN22 expression was reduced. Together, our results identified that PTPN22 is essential for THY1 to suppress cell invasion and SRC activity, maintain tight adherens junctions, and prevent NPC metastasis. These results suggested that PDGF-Rβ and SRC can be used as drug targets for suppressing NPC metastasis. Indeed, our in vivo assay using the SRC inhibitor KX2-391, clearly showed that inhibition of SRC signaling can prevent the metastasis of NPC, indicating that targeting SRC can be a promising approach to control the NPC progression.

A reversible SRC-relayed COX2 inflammatory program drives resistance to BRAF and EGFR inhibition in BRAFV600E colorectal tumors

BRAFV600E mutation confers a poor prognosis in metastatic colorectal cancer (CRC) despite combinatorial targeted therapies based on the latest understanding of signaling circuitry. To identify parallel resistance mechanisms induced by BRAF–MEK–EGFR co-targeting, we used a high-throughput kinase activity mapping platform. Here we show that SRC kinases are systematically activated in BRAFV600E CRC following targeted inhibition of BRAF ± EGFR and that coordinated targeting of SRC with BRAF ± EGFR increases treatment efficacy in vitro and in vivo. SRC drives resistance to BRAF ± EGFR targeted therapy independently of ERK signaling by inducing transcriptional reprogramming through β-catenin (CTNNB1). The EGFR-independent compensatory activation of SRC kinases is mediated by an autocrine prostaglandin E2 loop that can be blocked with cyclooxygenase-2 (COX2) inhibitors. Co-targeting of COX2 with BRAF + EGFR promotes durable suppression of tumor growth in patient-derived tumor xenograft models. COX2 inhibition represents a drug-repurposing strategy to overcome therapeutic resistance in BRAFV600E CRC.

Targeting Src reactivates pyroptosis to reverse chemoresistance in lung and pancreatic cancer models.

Pancreatic and lung cancers frequently develop resistance to chemotherapy-induced cell apoptosis during the treatment, indicating that targeting nonapoptotic-related pathways, such as pyroptosis, can be an alternative cancer treatment strategy. Pyroptosis is a gasdermin-driven lytic programmed cell death triggered by inflammatory caspases when initiated by canonical or noncanonical pathways that has been recently seen as a potential therapeutic target in cancer treatment. However, overcoming chemoresistance in cancers by modulating pyroptosis has not been explored. Here, we demonstrate that β5-integrin represses chemotherapy-induced canonical pyroptosis to confer cancer chemoresistance through ASAH2-driven sphingolipid metabolic reprogramming. Clinically, high β5-integrin expression associates with poor patient prognosis and chemotherapeutic responses in cancers. In addition, chemoresistant cells in vitro fail to undergo chemotherapy-induced pyroptosis, which is controlled by β5-integrin. Mechanistically, proteomic and lipidomic analyses indicate that β5-integrin up-regulates sphingolipid metabolic enzyme ceramidase (ASAH2) expression through Src-signal transducer and activator of transcription 3 (STAT3) signaling, which then reduces the metabolite ceramide concentration and subsequent ROS production to prohibit chemotherapy-induced canonical pyroptosis. Using cancer cell lines, patient-derived tumor organoids, and orthotopic lung and pancreatic animal models, we show that administration of a Src or ceramidase inhibitor rescues the response of chemoresistant pancreatic and lung cancer cells to chemotherapy by reactivating pyroptosis in vitro and in vivo. Overall, our results suggest that pyroptosis-based therapy is a means to improve cancer treatment and warrants further investigation.

Identification of potential bioactive phytochemicals, antioxidant properties and anticancer pathways of Terminalia ferdinandiana pulp

Fruits contain natural compounds which are useful for therapeutic effects. Phytochemicals are secondary metabolites created by plants during their biosynthetic pathways with a wide range of antifungal, antiviral, anticancer, and antibacterial activities. This research was carried out to examine the antioxidant properties and bioactive phytochemicals of Terminalia ferdinandiana, as well as to investigate its anticancer processes using a network pharmacology strategy. Several phytochemicals were found during GC-MS and LC-MS/MS investigation of non-hydrolyzed samples (with SPE, without SPE), and hydrolyzed (with SPE, without SPE) fractions. The GC-MS-based analysis identified the furoic acid, pentanoic acid, levulinic acid, p-hydroxy benzoic acid, gallic acid, and ellagic acid in the pulp extract. Moreover, LC-MS/MS-based investigation explored the presence of ellagic acid, chebulagic acid, chebulinic acid, resveratrol, catechin, p-coumaric acid, epicatechin, and gallic acid in the extract. By using the network pharmacology strategy, 100 target genes, including top hub genes associated with cancer-associated pathways were predicted. These targeted genes are mainly associated with cancer, cellular signaling, metabolism, and immunity-associated pathways. Based on the molecular docking approach, we revealed the bioactive phytochemicals that interact with hub target proteins, including AKT1, VEGFA, STAT3, SRC, and ESR1. Identification of bioactive phytochemicals and their predicted target genes may play crucial roles in the exact field of nutrigenomics, functional foods, and nutraceuticals from this natural fruit.

Elucidation of the potential molecular mechanism of the active compounds of Bryophyllum pinnatum (L. f.) Oken against gastritis based on network pharmacology

Bryophyllum pinnatum (L. f.) Oken is a kind of herbaceous plant of Crassulaceae, although studies have shown that Bryophyllum pinnatum can treat gastritis, the main active compounds and molecular mechanisms are still unclear. In this study, UPLC-MS analyses and a strategy of network pharmacology were used to elucidate the potential molecular mechanisms of Bryophyllum pinnatum against gastritis. 85 compounds in Bryophyllum pinnatum were identified through UPLC-MS analysis, 7 main active compounds (quercetin, eicosapentaenoic acid (EPA), isorhamnetin, beta-sitosterol, cubebininolide, kaempferol, artemetin) were obtained based on online database. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis demonstrated that 93 common targets were mainly enriched in pathways in cancer, PI3K-Akt signaling pathway, FoxO signaling pathway, TNF signaling pathway. AKT1, SRC, EGFR, MAPK1 and MAPK8 targets were the core targets. The molecular mechanisms of Bryophyllum pinnatum to exert anti-gastritis effect were mainly related to the control of cell physiological activities and reduction of stomach acids and gastric proteases. In summary, our study reveals that Bryophyllum pinnatum has the characteristics of multitarget, multi-compound and multi-pathway to exert anti-gastritis effect, which provides the direction and reference for the subsequent in-depth verificationof the mechanisms of anti-gastritis effect of Bryophyllum pinnatum ..

Bufalin inhibits epithelial-mesenchymal transition and increases radiosensitivity of non-small cell lung cancer via inhibition of the Src signaling.

Epithelial-mesenchymal transition (EMT) is a biological process involved in tumor migration, invasion, and radiotherapy resistance. Bufalin can affect the proliferation, apoptosis and invasion of tumor cells by regulating multiple signaling pathways. Whether bufalin can increase radiosensitivity through EMT deserves further investigation. In this study, we investigated the effect of bufalin on the EMT and radiosensitivity of non-small cell lung cancer (NSCLC) and the underlying molecular mechanism. NSCLC cells were treated with bufalin (at a dose of 0-100 nM) or irradiated with 6 MV X-rays (4 Gy/min). The effects of bufalin on cell survival, cell cycle, radiosensitivity, cell migration, and invasion were detected. Western blot was used to analyze the gene expression changes of Src signaling in NSCLC cell induced by Bufalin. Bufalin significantly inhibited cell survival, migration, and invasion and induced G2/M arrest and apoptosis. Cells co-treated with bufalin and radiation manifested a higher inhibitory effect compared to those treated with radiation or bufalin alone. Furthermore, the levels of p-Src and p-STAT3 were considerably reduced following bufalin treatment. Interestingly, elevated p-Src and p-STAT3 were observed in cells treated with radiation. Bufalin inhibited radiation-induced p-Src and p-STAT3, whereas the knockdown of Src abrogated the effects of bufalin on cell migration, invasion, EMT, and radiosensitivity. Bufalin inhibits EMT and enhances radiosensitivity through targeting Src signaling in NSCLC.