The saponin DT-13 inhibits gastric cancer cell migration through down-regulation of CCR5-CCL5 axis

AIM： To investigate the effect of DT-13 on gastric cancer cell migration, and to explore the possible mechanisms underlying the anti-metastasis activity of DT-13. METHODS： Growth inhibition of DT-13 was analyzed by the MTT assay. Cell migration was measured by the scratch-wound assay and transwell double chamber assay. To investigate the possible mechanisms underlying the anti-metastasis activity of DT-13, chemokine receptors that are involved in cancer metastasis（CCR2, CCR5, CCR7, CXCR4, and CXCR6） were detected by conventional PCR. The effect of DT-13 on CCR5 and CXCR4 expression was further evaluated by quantitative PCR and Western blot, respectively. The secretion of CCL5（ligand of CCR5） and SDF-1（ligand of CXCR4） were detected by enzyme-linked immunosorbent assay（ELISA）. RESULTS： DT-13 inhibited BGC-823 and HGC-27 cell growth in a dose dependent manner, and the estimated IC50 value for 24 h treatment was 23.5 ± 5.1 μmol·L^-1 for BGC-823 cells and 35.6 ± 7.6 μmol·L^-1 for HGC-27 cells. DT-13 also significantly decreased gastric cancer cell migration. DT-13 significantly decreased the gene expression of CCR5 in both BGC-823 and HGC-27 gastric cancer cells, and moderately reduced the expression of CXCR4. Similar to the results of gene expression, significant down-regulation of CCR5 protein was observed, but CXCR4 protein levels were much less affected. CCL5 secretion, but not SDF-1 production, was inhibited by DT-13. CONCLUSION： DT-13 inhibited gastric cancer cell migration by down-regulation of the CCR5-CCL5 axis.     

Exploration of Bioactive Constituents and Immunoregulatory Mechanisms of a Hanshi‑Yufei Formulation for Treating COVID‑19

Objective:The objective of this study was to characterize the chemical compounds of a Hanshi-Yufei formulation (HSYF; a modified formulation of a traditional Chinese medicine used for treating COVID-19) to elucidate the mechanism of action and to evaluate potential anti-inflammatory effects of HSYF. Materials and Methods:The chemical constituents of HSYF extract were characterized using UPLC-Q-TOF/MS.Subsequently,asetofTCMnetworkpharmacologymethodswasappliedtoidentifydisease-associatedgenesandtopredicttarget profiles and pharmacological actions associated with the constituents of HSYF. Then, the antiviral effects of HSYF on H1N1 were assessed in RAW264.7 cells using MTT assays. Expression levels of pro-inflammatory cytokines interleukin (IL)-6 and tumor necrosis factor (TNF)-αfollowing infection of RAW264.7 cells with H1N1 were measured using an enzyme-linked immune sorbent assay (ELISA), and expression levels of inflammatory-related factors were detected using western blotting. Results:In total, 165 chemical constituents (including glycosides,tannins, volatile oils, amino acids, triterpenoids, polyphenols, phenylpropanoids, sesquiterpenes, alkaloids, and flavonoids, among others) were tentatively identified in HSYF. Network pharmacology demonstrated that HSYF can regulate immunomodulatory-and anti-inflammatory-related targets of multiple pathways through its active ingredients, suggesting potential anti-COVID-19 effects. Furthermore, cell viability assays and ELISA showed that HSYF significantly inhibited H1N1 replication in RAW64.7 cells and markedly reduced expression of pro-inflammatory cytokines TNF-αand IL-6 at the proteins level. Conclusions:The results of the present study help improve our understanding of the therapeutic effects of HSYF in COVID-19 treatment from multi-level perspectives.     

The Total Flavonoids of Clerodendrum bungei Suppress A549 Cells Proliferation,Migration,and Invasion by Impacting Wnt/β-Catenin Signaling

Objectives: The objective of this study is to evaluate the effect of the total flavonoids of Clerodendrum bungei(TFCB) on the proliferation,invasion, and metastasis of A549 lung cancer cells through the Wnt signaling pathway. Materials and Methods: A549 cells were transfected with a β-catenin overexpression plasmid and the empty vector pcDNA3.1. The A549 cells were divided into six groups: normal A549 cell group, normal A549 cells with TFCB group, vector control group, vector with TFCB group,(3-catenin overexpression group, and β-catenin with TFCB group. We used the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay to detect cell proliferation, a scratch test was used to observe cell migration, and a transwell experiment was employed to evaluate cell invasion. Proteins related to the Wnt pathway were detected with Western blot analysis, including β-catenin, GSK-3 β, P-GSK-3 β, c-Myc, and CyclinD1. Results: The proliferation, invasion,and metastasis of A549 cells were significantly enhanced after being transfected with the β-catenin overexpression plasmid(P 0.05 or 0.01),accompanied by increased expression of β-catenin, C-Myc, CyclinD1 and reduced expression of Gsk-3 β and P-GSK-3 β. Treatment of cells with TFCB resulted in inhibition of cell proliferation, migration, and invasion; downregulated expression of β-catenin, C-Myc, and CyclinD1;and upregulated expression of GSK-3 β and P-GSK-3 β,especially in the β-catenin overexpression group. Conclusion: TFCB has the potential to inhibit the Wnt/β-catenin pathway by prohibiting the overexpression of β-catenin and regulating its downstream factors.     

Baicalin extracted from Huangqin(Radix Scutellariae Baicalensis) induces apoptosis in gastric cancer cells by regulating B cell lymphoma(Bcl-2)/Bcl-2-associated X protein and activating caspase-3 and caspase-9

OBJECTIVE:To evaluate the effects of baicalin in human gastric cancer cells, including apoptosis-inducing effects, and to investigate its underlying mechanisms of action.METHODS:Cell proliferation and apoptosis assays were performed to investigate the anti-proliferation effects of baicalin in human gastric cancer BGC-823 and MGC-803 cells.Real time-quantitative polymerase chain reaction and Western blotting analysis were performed to elucidate the molecular mechanisms underlying the anti-tumor properties of baicalin.RESULTS:In BGC-823 and MGC-803 gastric cancer cells treated with 80, 120, and 160 μmol/L baicalin for 48 h, a 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide assay showed that baicalin significantly inhibited cell proliferation in a dose-dependent manner, while flow cytometric analysis demonstrated that baicalin could induce apoptosis, also in a dose-dependent manner.Moreover, baicalin up-regulated the expression of caspase-3, caspase-9, and B cell lymphoma(Bcl-2)-associated X protein and down-regulated the expression of Bcl-2 at both the m RNA and protein level.CONCLUSION:Baicalin has potential as a therapeutic agent for gastric cancer by inducing apoptosis in cancer cells.     

Inhibition of Corydalis decumbens Alkaloids on Hydrogen Peroxide- induced Apoptosis of PC12 Cells through Down-regulating Caspase-3 Expression

Objective To extract alkaloids from Corydalis decumbens(AsCD) by supercritical CO2 fluid extraction(SFE) and to evaluate protective effects of AsCD against hydrogen peroxide(H2O2)-induced apoptosis in rat PC12 cells. Methods AsCD were extracted by SFE and oxidative damage PC12 cells model was induced by H2O2.The survival rate of the cells was determined by MTT assay;Lactate dehydrogenase release was determined by ultraviolet spectrophotometry;Flow cytometry was used to detect apoptosis;Caspase-3 mRNA and protein were determined by real-time PCR and Western blotting assay,respectively.Results AsCD remarkably reduced the cytotoxicity, prevented membrane damage,and inhibited cell apoptosis.AsCD inhibited Caspase-3 mRNA and protein expression induced by H2O2 in PC12 cells.Conclusion AsCD possess protective effects against H2O2-induced apoptosis in PC12 cells,and the mechanism of AsCD responsible to the inhibition of apoptosis is possibly attributed to the down-regulating Caspase-3 expression.AsCD might be useful in the treatment of oxidative stress-related neurodegenerative diseases.     

Electroacupuncture inhibits annulus fibrosis cell apoptosis in vivo via TNF-α-TNFR1-caspase-8 and integrin β1/Akt signaling pathways

OBJECTIVE： To examine whether electroacupuncture（EA） treatment inhibited cell apoptosis of intervertebral annulus fibrosis（AF） via tumor necrosis factor-α（TNF-α）-tumor necrosis factor receptor 1（TNFR1）-caspase-8 and integrin β1/Akt signaling pathways in a rat model of cervical intervertebral disc degeneration caused by unbalanced dynamic and static forces.METHODS： Thirty-two Sprague-Dawley rats were included in this study, of which 24 rats underwent surgery to induce cervical intervertebral disc degeneration, while eight rats received EA treatment at Dazhui（GV 14）. Immunohistochemical staining was used to detect TNF-α, TNFR1, and caspase-8Apoptosis of AF cells was examined with terminal deoxynucleotidyl transferase-mediated d UTP-biotin nick end labeling（TUNEL） staining. The m RNA and protein expression levels of integrin β1 andAkt were evaluated with real-time polymerase chain reaction and western blot analysis, respectively.RESULTS： Treatment with EA decreased TUNEL-positive AF cells and lowered TNF-α, TNFR1 and caspase-8 positive cells compared with control groups. EA treatment also increased integrin β1and Akt m RNA and protein levels compared with controls.CONCLUSION： Treatment with EA inhibits AF cell apoptosis through suppression of the TNF-α-TNFR1-caspase-8 signal pathway and increases the expression of integrin β1 and Akt. EA may be a good alternative therapy for treating cervical spondylosis.     

Fuzheng Kang'ai decoction(扶正抗癌方) inhibits cell proliferation,migration and invasion by modulating mir-21-5p/human phosphatase and tensin homology deleted on chromosome ten in lung cancer cells

OBJECTIVE: To elucidate the potential molecular mechanism by which Fuzheng Kang’ai decoction(扶正抗癌方, FZKA) inhibits proliferation, migration, and invasion of lung cancer cells. METHODS: Varying FZKA concentrations were used to manage lung cancer cells(A549 and PC9). We employed: cell counting kit-8(CCK-8) and plate clone formation assays to examine the cell viability; flow cytometry(FCM) to analyze the cycle arrest; transwell and woundhealing assays to assess the cell invasion and migration, resp...     

Ginsenoside 3β‑O‑Glc‑DM (C3DM) Enhances the Antitumor Activity of Taxol on Lewis Lung Cancer by Targeting the Interleukin‑6/Jak2/STAT3 and Interleukin‑6/AKT Signaling Pathways

Nonsmall?cell lung cancer (NSCLC) is an aggressive, highly chemoresistant disease. Taxol is an effective chemotherapeutic drug widely used for the treatment of NSCLC. However, the clinical use of Taxol is limited due to the occurrence of adverse side effects under high therapeutic doses. Therefore, it is desirable to explore combination therapy to reduce the dose of chemotherapeutic drugs and achieve excellent outcomes. A biosynthetic ginsenoside, 3-O-β-D-glucopyranosyl-dammar-24-ene-3β, 20S?diol (3β-O-Glc-DM, C3DM) is obtained from microbial fermentation by metabolic engineering. Based on previous study findings, we aimed to explore the mechanism of combination therapy with C3DM and Taxol and itsincreasing antitumor effect on Lewislung cancer (LLC) in thisstudy. Materials and Methods: Athiazolyl blue tetrazolium bromide (MTT) assay was performed to evaluate cell viability; the apoptotic effect was studied using cell apoptosis assay. The Lewis tumor xenograft experiment was performed to determine the effects of C3DM combined with Taxol on tumor growth in vivo, and western blotting was performed to analyze protein expressions. Results: C3DM effectively inhibited the proliferation of NSCLC cells. Moreover, C3DM increased the antiproliferative activity of Taxol and significantly enhanced cell apoptosis induced by Taxol in Lewis lung cancer cells. C3DM alone also suppressed Lewis tumor growth and enhanced the antitumor activity of Taxol in vivo. Western blot analysis revealed that the effects of the antiproliferation and apoptosis induction of C3DM treatment alone or in combination with Taxol on Lewis lung cancer were mediated by inhibiting the interleukin?6 (IL?6)/Jak2/STAT3 and IL?6/AKT signaling pathways. Conclusions: The results showed that C3DM has the potential to be used in combination therapy with Taxol against NSCLC.     

Pterostilbene supresses inflammation-induced melanoma metastasis by impeding neutrophil elastase-mediated thrombospondin-1 degradation

Objective: Chronic inflammation plays a fatal role in tumor metastasis. Pterostilbene (PTE) is a natural dimethylated analogue of resveratrol with anticancer and anti-inflammatory activities. This study aimed to investigate the inhibitory effect of PTE on inflammation-associated metastasis and explore the underlying mechanisms. Methods: Lipopolysaccharide (LPS)-induced lung inflammation and melanoma metastasis models were established in mice. After PTE treatment for four weeks, the organ index, histological changes, proinflammatory cytokines, and the expression and activity of neutrophil elastase (NE), a biomarker of neutrophil influx in the lungs, were analysed. Additionally, direct effects of PTE on NE-induced B16 cell migration were explored in wound healing and Transwell assays, and the expression of thrombospondin-1 (TSP-1) and epithelial-mesenchymal transition (EMT) markers were also detected. Results: PTE obviously attenuated the LPS-induced metastasis of circulatory B16 cells to lungs by reducing the number of metastatic nodules on the lung surfaces and the lung weight/body weight ratio. PTE treatment also significantly reduced LPS-activated increase levels of tumor necrosis factor (TNF)-α and interleukin (IL)-6 in the lungs of tumor-bearing mice. In addition, increased expression and enzyme activity of NE and decreased expression of TSP-1 were observed, and these were blocked by PTE. In vitro, PTE at concentrations without cytotoxicity also markedly suppressed NE-triggered B16 cell migration, prevented NE-induced TSP-1 proteolysis and reversed the expression of vimentin, N-cadherin and E-cadherin. Conclusion: PTE could block inflammation-enhanced tumor metastasis, and the underlying mechanism might be associated with the inhibition of NE-mediated TSP-1 degradation.     

Brusatol Inhibits Proliferation,Migration, and Invasion of Nonsmall Cell Lung Cancer PC-9 Cells

Objective: The purpose of this study was to investigate the inhibitory effect of brusatol, a nigakilactone extracted from Brucea javanica, on lung cancer for development of therapeutic drugs. We explored the effects of brusatol on the proliferation, migration, and invasion of lung cancer PC-9 cells in vitro and analyzed the mechanisms involved. Materials and Methods: MTT assay was used to determine the effect of brusatol on the proliferative capacity of PC-9 and H1975 cells in vitro. The half-ma...     

Meta‑analysis of Clinical Efficacy of Traditional Chinese Medicine in the Treatment of Aplastic Anemia

Background: The “Diagnosis and Treatment Guidelines for Chinese Medicine Internal Common Diseases” issued by the Chinese Institute of traditional Chinese medicine (TCM) offers many methods for the treatment of aplastic anemia (AA). However, there is a lack of corresponding evidence. Objective: The study aimed to evaluate the clinical efficacy of TCM in the treatment of AA, and provide evidence for the development of guidelines for the diagnosis and treatment of AA using TCM. Methods: Data of randomized or semi?randomized control trials of AA treatments with TCM were retrieved, and the selected literature was scored using the Jadad scale. The data were extracted, and RevMan 5.2.6 software was used for the meta?analysis. Results: Two studies on the treatment of AA using Liuwei Dihuang pills combined with compound Zaofan pill were included. The results of the meta?analysis showed that there were no statistically significant differences in the efficacy between Liuwei Dihuang pills combined with compound Zaofan pill and androgen in the treatment of AA (P = 0.65). However, there were less adverse reactions, including liver damage and the hirsutism of women, with the former than the latter (P < 0.05). Other studies on the treatment of AA with TCM did not include reports from clinical trials. Conclusion: TCM had a certain curative effect when used to treat AA. However, the quality of the literature is generally low, and the sample size is small, which makes the validation of the results poor. Further high?quality studies are needed to provide high?level evidence.     

Brucea javanica oil inhibits proliferation of hepatocellular carcinoma cells and induces apoptosis via the PI3K/AKT pathway

Background:Brucea javanica oil(BJO),distributed primarily in Southeast Asia,has long been utilized as a therapeutic agent for treating malignancies.However,its anticancer mechanisms are not clearly understood.The objective of this study was to examine the mechanisms underlying its treatment of hepatocellular carcinoma cells.Methods:CCK8 assay was used to evaluate cell viability.Hoechst33342 staining and flow cytometry analyses were used to examine apoptosis.Mito-Tracker Red CMXRos kit was used to measure the membrane potential of mitochondria.ATP assay kit was used to evaluate ATP levels.Western blots were used to assess the presence of AKT,adenosine monophosphate-activated protein kinase,Caspase3,Caspase9,Bax,and Bcl-2.Results:BJO inhibited the proliferation of hepatocellular carcinoma cells HepG2 in a time-and dose-dependent manner.It induced apoptosis,with the percentage of cells treated with 50–150μg/mL BJO increasing from 8.01%to 28.02%in a concentration-dependent manner(P<0.05,when 50μg/mL of BJO group compared with the control group;P<0.001,when 100 or 150μg/mL of BJO group compared with the control group).After exposed to BJO,the expression of C-caspase3,C-caspase9 and Bax upregulated while that of Bcl-2 downregulated.BJO suppressed the PI3K/AKT pathway and promoted phosphorylation of adenosine monophosphate-activated protein kinase,while repressing the phosphorylation of mechanistic target of rapamycin.Compared with treatment by BJO alone,the PI3K/AKT agonist 740Y-P increased the survival rate of HepG2 cells(P<0.01)and attenuated the inhibitory effect of BJO on cell apoptosis(P<0.05).Conclusion:BJO is capable of inhibiting proliferation of HepG2 cells and inducing apoptosis via the PI3K/AKT pathway.     

DT-13, a saponin of dwarf lilyturf tuber,exhibits anti-cancer activity by down-regulating C-C chemokine receptor type 5 and vascular endothelial growth factor in MDA-MB-435 cells简

AIM： To investigate the anticancer activity of DT-13 under normoxia and determine the underlying mechanisms of action. METHODS： MDA-MB-435 cell proliferation, migration, and adhesion were performed to assess the anticancer activity of DT-13, a saponin from Ophiopogonjaponicus, in vitro. In addition, the effects of DT-13 on tumor growth and metastasis in vivo were evaluated by orthotopic implantation of MDA-MB-435 cells into nude mice; mRNA levels of vascular endothelial growth factor （VEGF）, C-C chemokine receptor type 5 （CCR5） and hypoxia-inducible factor 1a （HIF-1a） were evaluated by real-time quantitative PCR; and CCR5 protein levels were detected by Western blot assay. RESULTS： At 0.01 to 1 umol·L -1, DT-13 inhibited MDA-MB-435 cell proliferation, migration, and adhesion significantly in vitro. DT-13 reduced VEGF and CCR5 mRNAs, and decreased CCR5 protein expression by down-regulating HIF-1 a. In addition, DT-13 inhibited MDA-MB-435 cell lung metastasis, and restricted tumor growth slightly in vivo. CONCLUSION： DT-13 inhibited MDA-MB-435 cell proliferation, adhesion, and migration in vitro, and lung metastasis in vivo by reducing VEGF, CCR5, and HIF-la expression.