The up-regulation of miR-221-3p by tumor-associated macrophages may be involved in the metastasis of prostate cancer / 现代泌尿外科杂志

【Objective】 To investigate the mechanism by which the up-regulation of miR-221-3p by tumor-associated macrophages (TAMs) may be involved in promoting the malignant metastasis of prostate cancer (PCa). 【Methods】 The microRNAs (miRNAs) expression profiles of 6 cases of metastatic PCa tissues were sequenced and analyzed.The primary TAMs were isolated.The expression of miR-221-3p was determined with qPCR.The miR-221-3p mimic or miR-221-3p inhibitor was transfected into RAW264.7 macrophages in vitro, and co-cultured with human prostate cancer PC3 cells.The proliferation, apoptosis, invasion and migration of PC3 cells were detected with CCK-8, flow cytometry (FCM), Transwell assay, respectively.Expressions of epithelial-mesenchymal transformation (EMT) related protein factors were determined with Western blot. 【Results】 In the 6 cases of metastatic PCa, hsa-miR-221-3p was significantly up-regulated in TAMs-derived from PCa tissues with positive lymph node metastasis (P<0.05).In the co-cultured system, compared with Mimic-NC group, miR-221-3p mimic group had significantly up-regulated proliferation, migration, invasion and EMT-related protein factors (except E-Cadherin) (P<0.05).Compared with Inhibitor-NC group, miR-221-3p inhibitor group had significantly up-regulated apoptosis rate, but down-regulated proliferation, migration, invasion and EMT-related protein factors (except E-Cadherin) (P<0.05). 【Conclusion】 The miR-221-3p expression up-regulate by TAMs may participate in the malignant metastasis of prostate cancer.

Mechanism of exosome in the pathogenesis of prostate cancer and its potential diagnostic application / 现代泌尿外科杂志

Prostate cancer (PCa) is one of the most common tumors in men.In recent years, various researches on this disease and clinical applications have benefited patients.Exosome is a subclass of extracellular vesicles (EVs).Many studies have explored the mechanisms of exosome in mediating epithelial mesenchymal transformation, angiogenesis, tumor microenvironment establishment, immune escape and drug resistance acquisition in PCa, which provides a new perspective for finding new diagnostic markers.This article reviews the role of exosome in the pathogenesis of PCa and its diagnostic application.

Different calcium ion concentrations affect epithelial mesenchymal transformation of human peritoneal mesothelial cells via endoplasmic reticulum stress / 中华危重病急救医学

Objective:To study the effects of different calcium ion concentrations on epithelial mesenchymal transformation (EMT) of human peritoneal mesothelial cell (HPMC) via endoplasmic reticulum stress (ERS).Methods:HPMC cell line HMrSV5 was cultured in vitro and treated in groups. The cells in the control group, high calcium group 1, and high calcium group 2 were treated with medium containing calcium ion concentrations of 1.25, 1.75, and 2.25 mmol/L, respectively. The solvent control group was treated with medium containing 1.25 mmol/L physiological calcium ion concentration and 0.1% dimethyl sulfoxide (DMSO), the high calcium+solvent group was treated with medium containing 2.25 mmol/L calcium ion concentration and 0.1% DMSO, the high calcium+4-phenylbutyric acid (4-PBA) group was treated with medium containing 2.25 mmol/L calcium ion concentration and 1 mmol/L ERS inhibitor 4-PBA, and each group was treated for 48 hours. Morphological changes of cells in each group were observed under light microscope. The expressions of epithelial cell phenotype marker zonula occluden-1 (ZO-1) and mesenchymal cell phenotype marker α-smooth muscle actin (α-SMA) in the cells were observed by immunofluorescence staining. The expressions of EMT marker genes E-cadherin, ZO-1, α-SMA and Vimentin were detected by fluorescence quantitative polymerase chain reaction (PCR). The expressions of ERS marker proteins phosphorylated protein kinase R-like endoplasmic reticulum kinase (p-PERK), phosphorylated eukaryotic initiation factor 2α (p-eIF2α), transcription activating factor 4 (ATF4) and C/EBP homologous protein (CHOP) were detected by Western blotting. Results:Compared with the control group, the morphology of HMrSV5 cells became slender and fibrotic, the fluorescence intensity of ZO-1 increased, and the fluorescence intensity of α-SMA decreased in high calcium 1 and high calcium 2 groups, indicating that the cells transformed from epithelial cells to mesenchyme cells. The mRNA expressions of E-cadherin and ZO-1 were significantly decreased, while the mRNA expressions of α-SMA and Vimentin and the protein expressions of p-PERK, p-eIF2α, ATF4 and CHOP were significantly increased, moreover, the expressions of the above marker genes or proteins in the high calcium 2 group was more obvious than those in the high calcium 1 group [E-cadherin mRNA (2 -ΔΔCt): 0.53±0.05 vs. 0.75±0.09, ZO-1 mRNA (2 -ΔΔCt): 0.42±0.06 vs. 0.69±0.06, α-SMA mRNA (2 -ΔΔCt): 1.81±0.16 vs. 1.32±0.14, Vimentin mRNA (2 -ΔΔCt): 2.05±0.22 vs. 1.48±0.16, p-PERK protein (p-PERK/β-actin): 0.81±0.09 vs. 0.59±0.06, p-eIF2α protein (p-eIF2α/β-actin): 0.87±0.10 vs. 0.50±0.06, ATF4 protein (ATF4/β-actin): 0.93±0.10 vs. 0.72±0.06, CHOP protein (CHOP/β-actin): 0.79±0.09 vs. 0.46±0.04, all P < 0.05]. Compared with the solvent control group, the morphological changes of cells, the expressions of EMT marker genes and ERS marker proteins after high calcium ion concentration of 2.25 mmol/L were consistent with those in the high calcium 2 group than control group. Compared with the high calcium+solvent group, the cell morphology recovered the characteristics of polygonal and pebble-like epithelial cells in the high calcium+4-PBA group, the fluorescence intensity of ZO-1 increased, the fluorescence intensity of α-SMA decreased, and the mRNA expressions of E-cadherin and ZO-1 in the cells were significantly increased [E-cadherin mRNA (2 -ΔΔCt): 0.86±0.09 vs. 0.57±0.04, ZO-1 mRNA (2 -ΔΔCt): 0.81±0.06 vs. 0.48±0.05, both P < 0.05], the mRNA expressions of α-SMA and Vimentin and the protein expressions of p-PERK, p-eIF2α, ATF4 and CHOP were significantly decreased [α-SMA mRNA (2 -ΔΔCt): 1.21±0.13 vs. 1.77±0.15, Vimentin mRNA (2 -ΔΔCt): 1.30±0.14 vs. 1.94±0.20, p-PERK protein (p-PERK/β-actin): 0.38±0.04 vs. 0.92±0.11, p-eIF2α protein (p-eIF2α/β-actin): 0.34±0.05 vs. 1.05±0.13, ATF4 protein (ATF4/β-actin): 0.57±0.06 vs. 0.97±0.11, CHOP protein (CHOP/β-actin): 0.51±0.04 vs. 0.90±0.12, all P < 0.05]. Conclusion:High calcium ion concentrations of 1.75 mmol/L and 2.25 mmol/L promote EMT of HPMC via activating ERS.

Improvement of Lung Function in Rats with Idiopathic Pulmonary Fibrosis by Shengxiantang via Regulating Cell Senescence Mediated by Wnt3a/β-catenin Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo observe the effect of Shengxiantang （SXT） on cell senescence mediated by wingless/integrated （Wnt）3a/β-catenin pathway in rats with idiopathic pulmonary fibrosis （IPF） and reveal the possible mechanism in improving lung function of IPF rats. MethodA total of 32 SPF level SD rats were randomly divided into sham group， model group， pirfenidone group， and SXT group. The IPF rat model was established by intratracheal instillation of bleomycin （0.005 g·kg-1）. The following day after surgery， rats in the SXT group were given the aqueous solution of SXT granules （0.78 g·kg-1）， and the pirfenidone group was given pirfenidone suspension （0.05 g·kg-1）. The other groups were given deionized water （10 mL·kg-1） for 28 consecutive days. Lung tissue was collected after the lung function was measured. The pathological changes of the lung tissue were observed by hematoxylin-eosin （HE） and Masson staining， and then the Szapiel score and Ashcroft score were performed. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to detect telomere length. Western blot was applied to detect the expressions of epithelial-mesenchymal transformation （EMT） markers ［α-smooth muscle actin （α-SMA） and E-cadherin］， telomere reverse transcriptase （TRET）， aging-related proteins （p53 and p21）， senescence-associated secretory phenotype ［interleukin-6 （IL-6） and matrix metalloproteinase-1 （MMP-1）］， and key proteins of Wnt signaling pathway ［Wnt3a， glycogen synthase kinase-3β （GSK-3β）， β-catenin， Cyclin D1， and c-Myc］. ResultCompared with those in the Sham group， peak expiratory flow （PEF） and minute ventilation volume （MV） in the model group were significantly decreased （P<0.01）， and the frequency of respiratory （f） was significantly increased （P<0.01）. The Szapiel score， Ashcroft score， and protein expression of α-SMA， p53， p21， IL-6， MMP-1， Wnt3a， GSK3β， β-catenin， Cyclin D1， and c-Myc were increased （P<0.01）. The expressions of E-cadherin and TERT， as well as telomere length were significantly decreased （P<0.01）. Compared with those in the model group， PEF and MV in the SXT group were significantly increased （P<0.01）， while f was significantly decreased （P<0.01）. The Szapiel score， Ashcroft score， and protein expression of α-SMA， p53， p21， IL-6， MMP-1， Wnt3a， GSK3β， β-catenin， Cyclin D1， and c-Myc were significantly decreased （P<0.05， P<0.01）. Nevertheless， the expression of E-cadherin and TERT， as well as telomere length were significantly increased （P<0.01）. ConclusionSXT presents a significant protective effect on lung function in IPF rats， and the prescription may act on the Wnt3a/β-catenin signaling pathway to regulate cell senescence induced by TERT to inhibit EMT.

Expression of MMP-9 in patients with chronic rhinosinusitis and its correlation with epithelial-mesenchymal transition / 天津医药

Objective To investigate the mechanism of matrix metalloproteinase(MMP)-9 involved in epithelial mesenchymal transformation(EMT)in chronic sinusitis(CRS).Methods The expression of MMP-9 from polypoid middle turbinate tissue was detected by immunohistochemical staining qPCR and Western blot assay in 42 patients with CRS and 8 patients underwent septoplasty.Primary human nasal epithelial cells HNEpc were cultured in vitro and divided into the control group,the TGF-β1 group(5 μg/L TGF-β1 intervention)and the TGF-β1+si-MMP-9 group(transfected with si-MMP-9 and 5 μg/L TGF-β1 intervention).The expression of MMP-9 was detected by cell immunofluorescence staining.Expression levels of TGF-β1,MMP-9 and EMT-related proteins E-cadherin,vimentin and α-SMA were detected by Western blot assay.Results(1)The positive expression rate of MMP-9 was significantly higher in the nasal mucosa of CRS with nasal polyps(CRSwNP)group(54.5%,12/22)than that of the CRS without polyps(25.0%,5/20)group and the control group(12.8%,1/8).The relative expression levels of MMP-9 mRNA and protein in nasal mucosa were higher in the CRSwNP group than those in the CRSsNP group and the control group(P＜0.05).(2)Compared with the control group,the expressions levels of TGF-β1,MMP-9,vimentin and α-SMA were increased in the TGF-β1 group,while the expression of E-cadherin was decreased(P＜0.05).Compared with the TGF-β1 group,expression levels of TGF-β1,MMP-9,vimentin and α-SMA were decreased in the TGF-β1+si-MMP-9 group,and the expression of E-cadherin was increased(P＜0.05).Conclusion The expression of MMP-9 is increased in CRS patients,which may be involved in the development of CRS through the regulation of EMT.

Mechanism Study of Soufeng Yuchuan Formula on Regulating Airway Remodeling in Asthmatic Rats Based on Epithelial-Mesenchymal Transformation / 中药新药与临床药理

Objective To explore the mechanism of Suofeng Yuchuan Formula on inhibiting airway remodeling in asthma by regulating epithelial-mesenchymal transformation(EMT).Methods Sixty rats were randomly divided into normal control group,model control group,Dexamethasone group,high-,medium-and low-dose groups of Soufeng Yuchuan Formula,with 10 rats in each group.In addition to the normal control group,the other groups were injected and inhaled with ovalbumin(OVA)to replicate the asthma rat model.The rats in each group were killed after 21 days of administration of the corresponding drug.Hematoxylin-eosin(HE)staining was used to observe the pathological changes of lung tissue.The ultrastructure of epithelial cells in rat lung was observed by transmission electron microscope.The contents of transforming growth factor β1(TGF-β1)and interleukin-17(IL-17)in serum were detected by enzyme-linked immunosorbent assay(ELISA).Real-time fluorescence quantitative polymerase chain reaction(RT-PCR)was used to detect mRNA expressions of TGF-β1,α-smooth muscle actin(α-SMA),E-cadherin and N-cadherin in lung homogenate.Protein expression levels of TGF-β1,α-SMA,E-cadherin and N-cadherin in lung tissue were detected by Western Blot.Results Compared with the normal control group,the serum contents of TGF-β1 and IL-17 in the model control group were significantly increased(P＜0.01).The mRNA and protein expressions of TGF-β1,α-SMA and N-cadherin in lung tissues were significantly up-regulated(P＜0.01),while the mRNA and protein expressions of E-cadherin were significantly down-regulated(P＜0.01).HE staining showed thickening of airway wall and alveolar wall,and a large number of inflammatory cells infiltrated in the visual field,and the inflammation score was significantly increased(P＜0.01).Transmission electron microscopy showed that the epithelial cells of lung tissue had severe edema.Local dissolution and thinning of intracellular matrix,and obvious swelling of organelles were observed.Compared with model control group,the contents of TGF-β1 and IL-17 in serum of dexamethasone group and all doses of Soufeng Yuchuan Formula groups were significantly decreased(P＜0.05,P＜0.01).The mRNA and protein expressions of TGF-β1,α-SMA and N-cadherin were significantly down-regulated(P＜0.05,P＜0.01),while the mRNA and protein expressions of E-cadherin were significantly up-regulated(P＜0.05,P＜0.01).The inflammatory cell infiltration of lung tissue and thickening of bronchial wall and alveolar wall were found to be reduced.The inflammation score of dexamethasone group and high-dose group of Suofeng Yuchuan Formula was reduced(P＜0.05).Ultrastructure showed that the degree of edema of epithelial cells was significantly reduced,the cell membrane was intact,the intracellular matrix was uniform,and most of the organelles were slightly swollen.Conclusion Soufeng Yuchuan Formula can reduce the level of TGF-β1,thereby improve EMT in lung tissue,and achieve the purpose of preventing and treating airway remodeling in asthma.

Involvement of let-7c in alveolar macrophage M2 polarization in coal mine dust-induced epithelial-mesenchymal transition in rat lung tissue / 环境与职业医学

Background Coal workers' pneumoconiosis (CWP) is a serious occupational lung disease and one of the prescript occupational diseases in China. Epithelial-mesenchymal transition (EMT) is involved in the diffuse fibrosis of lung tissue of pneumoconiosis patients, and its mechanism may be related to the polarization of macrophages regulated by let-7c. Objective To investigate the effect of let-7c on the regulation of macrophage polarization in EMT in rats induced by coal dust exposure with different content of free SiO2. Methods SD rats were randomly divided into a control group, a 5% SiO2 group, a 30% SiO2 group, and a 99.9% SiO2 group, with 16 rats in each group. The rats in each group were tracheally titrated with 100 μL of 20 mg·mL−1 suspension (5% SiO2, 30% SiO2, and 99.9% SiO2) or normal saline, respectively. Alveolar lavage fluid was collected at the ends of the 1st month and the 3rd month. The relative expression levels of M1 or M2 markers, CD86 or CD206, in alveolar macrophages (AMs) were detected by immunofluorescence. The inflammation and fibrosis of lung tissue were observed by hematoxylin-eosin (HE) staining and Masson staining. The expression levels of transforming growth factor-β1 (TGF-β1), E-cadherin, and vimentin were detected by Western blotting. The expression levels of let-7c and c/EBP-δ genes were detected by real-time fluorescence quantitative PCR. Results The HE and Masson staining results showed that compared with the control group, the degree of pulmonary fibrosis in the 5% SiO2 group, the 30% SiO2 group, and the 99.9% SiO2 group gradually increased with the increase of dust exposure time. Compared with the control group, the expressions of CD86 and CD206 in the 5% SiO2 group, the 30% SiO2 group, and the 99.9% SiO2 group gradually increased at the end of the 1st month (F=330.904, 146.801, P<0.05), and the expression of CD86 in each group decreased gradually at the end of the 3rd month (F=331.781, P<0.05), but the expression of CD206 increased (F=1164.190, P<0.05). At the end of the 1st month, the expressions of TGF-β1 (F=8.847, P<0.05) and vimentin (F=13.275, P<0.05) gradually increased, and the expression of E-cadherin (F=6.253, P<0.05) gradually reduced in the 5% SiO2 group, the 30% SiO2 group, and the 99.9% SiO2 group. At the end of the 3rd month, the expressions of TGF-β1 (F=16.833, P<0.05) and vimentin (F=55.021, P<0.05) increased, and the expression of E-cadherin (F=12.790, P<0.05) gradually decreased in all groups. The PCR results showed that compared with the control group, the expression of let-7c mRNA in the 5% SiO2 group, the 30% SiO2 group, and the 99.9% SiO2 group increased at the ends of the 1st month and the 3rd month (F=11.251, 28.136, P<0.05). The expression of c/EBP-δ mRNA decreased in all groups at the ends of the 1st month and the 3rd month (F=49.204, 177.090, P<0.05). Conclusion In response to dust stimulation, let-7c promotes EMT by modulating macrophage polarization, which is involved in the formation of pulmonary fibrosis and thus influences the progression of CWP .

miR-186 regulates epithelial-mesenchymal transformation to promote nasopharyngeal carcinoma metastasis by targeting ZEB1

Abstract Objectives Nasopharyngeal carcinoma (NPC) is an aggressive epithelial cancer. The expression of miR-186 is decreased in a variety of malignancies and can promote the invasion and metastasis of cancer cells. This study aimed to explore the role and possible mechanism of miR-186 in the metastasis and epithelial-mesenchymal transformation (EMT) of NPC. Methods The expression of miR-186 in NPC tissues and cells was detected by RT-PCR. Then, miR-186 mimic was used to transfect NPC cell lines C666-1 and CNE-2, and cell activity, invasion and migration were detected by CCK8, transwell and scratch assay, respectively. The expression of EMT-related proteins was analyzed by western blotting analysis. The binding relationship between miR-186 and target gene Zinc Finger E-Box Binding Homeobox 1 (ZEB1) was confirmed by double luciferase assay. Results The expression of miR-186 in NPC was significantly decreased, and transfection of miR-186 mimic could significantly inhibit the cell activity, invasion, and migration, and regulate the protein expressions of E-cadherin, N-cadherin and vimentin in C666-1 and CNE-2 cells. Further experiments confirmed that miR-186 could directly target ZEB1 and negatively regulate its expression. In addition, ZEB1 has been confirmed to be highly expressed in NPC, and inhibition of ZEB1 could inhibit the activity, invasion, metastasis and EMT of NPC cells. And co-transfection of miR-186 mimic and si-ZEB1 could further inhibit the proliferation and metastasis of NPC. Conclusion miR-186 may inhibit the proliferation, metastasis and EMT of NPC by targeting ZEB1, and the miR-186/ZEB1 axis plays an important role in NPC.

Effect of long non-coding RNA 114227 on gastric cancer cell proliferation and migration / 中南大学学报(医学版)

OBJECTIVES@#Gastric cancer is a common cancer of the digestive system. Long non-coding RNA (lncRNA) plays an important role in the formation and development of gastric cancer. This study aims to investigate the effect of long non-coding lncRNA 114227 on biologic behaviors in gastric cancer cells.@*METHODS@#The experiment was divided into 4 groups: a negative control (NC) group, a lncRNA 114227 small interference (si-lncRNA 114227) group, an empty vector (Vector) group, and an overexpression vector (OE-lncRNA 114227) group. The expressions of lncRNA 114227 in gastric mucosa and gastric cancer tissues, gastric mucosal epithelial cells and different gastric cancer strains were determined by real-time reverse transcription PCR (real-time RT-PCR).The proliferation were detected by CCK-8 assay in gastric cancer cells. The epithelial-mesenchymal transformation (EMT) was utilized by Transwell assay, scratch healing assay, and Western blotting in gastric cancer cells. The effect of lncRNA 114227 on proliferation of gastric cancer cells was detected by tumor bearing experiment in nude mice in vivo.@*RESULTS@#The expression level of lncRNA 114227 in the gastric cancer tissues was significantly lower than that in the gastric mucosa tissues, and in 4 kinds of gastric cancer strains was all significantly lower than that in gastric mucosal epithelial cells (all P<0.01). In vitro, the proliferation and migration abilities of gastric cells were significantly reduced after overexpressing lncRNA 114227, and cell proliferation and migration were enhanced after silencing lncRNA 114227 (all P<0.05). The results of in vivo subcutaneous tumorigenesis in nude mice showed that the tumorigenic volume of the tumor-bearing mice in the OE-lncRNA 114227 group was significantly smaller than that of the Vector group, and the tumorigenic quality was lower than that of the Vector group (P<0.05), indicating that lncRNA 114227 inhibited tumorigenesis.@*CONCLUSIONS@#The expression of lncRNA 114227 is downregulated in gastric cancer gastric cancer tissues and cell lines. LncRNA 114227 may inhibit the proliferation and migration of gastric cancer cells through EMT process.

Screening of EMT-related genes in lung adenocarcinoma and construction of ceRNA network / 国际检验医学杂志

Objective To screen the epithelial mesenchymal transformation(EMT)-related genes in lung adenocarcinoma,perform functional enrichment analysis and construct protein interaction network(PPI).Ac-cording to the competitive endogenous RNA(ceRNA)hypothesis and the effect of gene expression on the prognosis of patients,the ceRNA network was constructed.Methods The differentially expressed genes be-tween lung adenocarcinoma tissues and normal tissues were screened by gene expression map and tumor ge-nome map database,and the genes were imported into GenClip3 to obtain EMT-related genes.Metascape was used to perform gene ontology and Kyoto encyclopedia of gene and genome enrichment analysis,and STRING database was used to construct PPI and obtain EMT key genes.The relationship between key genes and prog-nosis was analyzed by Kaplan-Meier analysis.Analysis tools such as miRTarbase,miRNet database,and EN-CORI were used to construct ceRNA networks.Results In this study,156 lung adenocarcinoma EMT-related genes and their key genes cadherin 1,interleukin-6,matrix metalloproteinase-9,platelet endothelial cell adhe-sion molecule,cyclin-dependent kinase inhibitor 2A,α1-Ⅰcollagen gene,secreted phosphoprotein 1,TIMP in-hibitor of matrix metalloproteinase-1,caveolin-1 and Zeste homologue enhance core 2(EZH2)-1 were identi-fied.The PPI of key genes was predicted,and the therapeutic drugs targeting these key genes including salvia miltiorrhiza,ginseng lu,ginseng leaf and ginseng flower were also predicted.The prognostic ceRNA regulatory network of EZH2/hsa-miR-101-3p/GSEC was constructed.Conclusion This study describes using bioinfor-matics methods system in the process of EMT gene interactions,according to the lung adenocarcinoma pa-tients clinical data to construct the prognosis of the EMT process related ceRNA network,for the treatment and prognosis of lung adenocarcinoma judgment provides a new way of thinking.

Study on Jianpi Bushen Jiedu Decoction for inhibiting epithelial-mesenchymal transformation of hepatocellular carcinoma through JAK2/STAT3 pathway / 国际中医中药杂志

Objective:To study the effects of Jianpi Bushen Jiedu Decoction on the epithelial-mesenchymal transformation of nude mice with HCCLM3 subcutaneous transplanted tumor by regulating JAK2/STAT3 pathway.Methods:HCCLM3 subcutaneous transplanted tumor model was established in mice. After the successful modeling, 24 nude mice were divided into blank group, TCM group and combined group according to random number table method, with 8 mice in each group. Mice in the TCM group were given 0.68 mg/ml alcohol extract of Jianpi Bushen Jiedu Decoction for gavage, and the combined group were given sorafenib suspension plus alcohol extract of Jianpi Bushen Jiedu Decoction 3.5 mg/ml for gavage, once a day, for consecutive 4 weeks. The effects of Jianpi Bushen Jiedu Decoction on tumor volume, tumor weight of HCCLM3 subcutaneous transplanted tumor and mice body weight were observed; Western blot was used to detect the expressions of E-cadherin, N-cadherin, Vimentin and JAK2/STAT3 pathway-related proteins in subcutaneous transplanted tumor tissues of hepatocellular carcinoma of mice in each group.Results:Compared with the control group, the average tumor weight of subcutaneous transplanted tumor decreased significantly in the TCM group and the combined group ( P<0.05), and the expressions of JAK2, STAT3, p-JAK2, p-STAT3, N-cadherin, and Vimentin decreased significantly in subcutaneous transplanted tumor tissue ( P<0.05), while E-cadherin increased ( P<0.05). Conclusion:Jianpi Bushen Jiedu Decoction can inhibit the growth of subcutaneous transplanted tumor of hepatocellular carcinoma in mice. The mechanism may be related to inhibiting the activation of JAK2/STAT3 pathway, thereby inhibiting the epithelial-mesenchymal transformation of hepatocellular carcinoma.

Study on Huangqi Jiedu Decoction for Wnt/β-catanin signaling pathway inhibiting EMT against lung metastasis of breast cancer / 国际中医中药杂志

Objective:To observe the inhibitory effects of Huangqi Jiedu Decoction on lung metastasis of breast cancer in nude mice; To explore the mechanism of intervening epithelial mesenchymal transformation (EMT) induced by Wnt/β-catenin signaling pathway.Methods:Totally 30 nude mice were divided into model group, adriamycin group and Huangqi Jiedu Decoction low-, medium-, and high-dosage groups according to random number table method. Each group was injected subcutaneously with mouse breast cancer 4T1 cells to construct tumor - bearing nude mice model. Huangqi Jiedu Decoction low-, medium- and high-dosage groups were intragastrically administrated with Huangqi Jiedu Decoction 17.82, 35.64 and 71.28 g/kg; adriamycin group was injected intraperitoneally adriamycin 0.05 g/kg; model group was intragastrically administrated with normal saline of the same volume for 21 d. Tumor volume was measured at 9, 15, and 21 days after modeling. After the end of administration, the tumor tissue was separated, the tumor weight was measured, and the tumor inhibition rate was calculated. The lung tissue was Isolated,, the number of lung metastatic nodules and the inhibition rate of lung metastasis was counted. HE staining was used to observe the tissue morphology and evaluate the effectiveness of the model. The protein expressions of β-catenin, E-Cadherin and Vimentin in lung tissue were detected by Western Blot. The mRNA levels of β-catenin, E-Cadherin and Vimentin in lung tissue were detected by real-time fluorescent quantitative PCR.Results:Compared with the model group, the tumor volume and mass of Huangqi Jiedu Decoction low-, medium- and high-dosage groups decreased ( P<0.01); the number of pulmonary metastasis nodules in Huangqi Jiedu Decoction high-dosage group significantly decreased ( P<0.01); the mRNA and protein expressions of β-catenin and Vimentinm decreased in the Huangqi Jiedu Decoction low-, medium- and high-dosage groups ( P<0.01), and the protein and mRNA expressions of E-Cadherin increased in the Huangqi Jiedu Decoction high-dosage group ( P<0.01). Conclusion:Huangqi Jiedu Decoction can effectively inhibit the growth and lung metastasis of breast cancer transplanted tumor, and the mechanism may be to down-regulate the expression of key molecules in the Wnt/β-catanin signaling pathway, thereby inhibiting the EMT process, so as to inhibit the lung metastasis of breast cancer.

Rab25 gene down-regulates snail expression and inhibits epithelial mesenchymal transformation of human breast cancer MDA-MB-231 cells / 中华内分泌外科杂志

Objective:To probe into Rab25 Gene’s Effect on TGF-β inhibition of proliferation, invasion and epithelial mesenchymal transformation (EMT) of breast cancer MDA-MB-231 cells and explore its molecular mechanism.Methods:The experiment was divided into three groups: control group,TGF-β Group and si-Rab25 group. TGF-β induced MDA-MB-231 cell model of EMT was built. CCK-8 assay was used to detect cell proliferation. Transwell assay was used to detect the ability of cell invasion and migration.Western blot was used to detect the changes of related proteins in each group.Results:After stimulating MDA-MB-231 cells with TGF-β, Rab25 gene was highly expressed. Compared with TGF-β group (57.48±%3.62%), the migration ability and invasion ability of cells in si-Rab25 group (33.49%±2.93%) decreased by 41.7%, with a significant difference ( P<0.05). Compared with TGF-β group (153.21%±4.17%), the proliferation ability of cells in si-Rab25 group (115.32%±5.69%) decreased by 24.73%, with a significant difference ( P<0.05). The expression of MDA-MB-231 fine EMT related protein in si-Rab25 group was significantly different from that in TGF-β group ( P<0.05). The expression of p-AKT and Snail protein in si-Rab25 group was significantly lower than that in TGF-β group ( P<0.05) . Conclusions:Rab25 gene is highly expressed in MDA-MB-231 cells. Silencing Rab25 gene can activate AKT signal pathway, inhibit Snail protein expression, regulate EMT related protein expression, and inhibit EMT transformation.

Advances in studies on the effect of epithelial-mesenchymal transformation on radiotherapy sensitivity of tumors / 中华放射肿瘤学杂志

Radiotherapy is one of the most important methods in the treatment of malignant tumors. However, the decrease of radiosensitivity of tumor cells is the main reason affecting the efficacy of radiotherapy. Epithelial-mesenchymal transition (EMT) is a complex biological process that confers several characteristics necessary for the progression of malignant tumors, such as tumor initiation, aggressiveness, transmissibility, and tolerance to chemotherapy and radiotherapy. In addition, EMT can also be induced by radiation, which endows tumor cells with radiation resistance. Previous studies have shown that inhibition of EMT could enhance the radiosensitivity of tumor cells, but the overall understanding of the molecular mechanisms, key targets and pathways involved are still lacking. In this article, recent studies on the role of EMT in tumor radiation therapy were reviewed, focusing on the signaling pathway, EMT-induced transcription factors, aiming to deepen the understanding of the effect of EMT on the sensitivity of radiotherapy and provide ideas for improving the clinical therapeutic effect of radiotherapy.

Expressions and clinical significance of ZEB1, AR, E-Ca and N-Ca in prostate cancer / 现代泌尿外科杂志

【Objective】 To analyze the correlation between the expressions of ZEB1, androgen receptor (AR), E-cadherin (E-Ca), N-cadherin (N-Ca) and clinicopathological features of prostate cancer patients with different risk levels, and to explore their significance. 【Methods】 The clinical data of 47 patients with prostate cancer treated during Nov.2013 and Jun.2021 were retrospectively analzyed. The patients were divided into medium-low risk group and high-risk group. The expressions of ZEB1, AR, E-Ca and N-Ca in the prostate cancer tissues of the two groups were detected with immunohistochemical staining. The relationship between the expressions and Gleason grade, prostate-specific antigen (PSA) level and TNM stage was analyzed. 【Results】 The positive expression rate of ZEB1 increased with higher risk, Gleason score, and PSA level (P<0.01); the strong positive expression rate of AR decreased with higher risk and Gleason score (P<0.05); the positive expression rate of E-Ca decreased with increased risk, Gleason score, and PSA level (P<0.05); the positive expression rate of N-Ca increased with the increased risk and Gleason score (P<0.01); the positive expression rate of ZEB1 increased with higher tumor stage and TNM stage (all P<0.01); the strong positive expression rate of AR decreased only with increased TNM stage (P<0.05). Patients whose first surgical specimen showing a higher expression level of ZEB1 were more likely to develop into castration-resistant prostate cancer CRPC (P<0.05). 【Conclusion】 ZEB1 and N-Ca levels increase with increased tumor aggressiveness, while AR and E-Ca levels decrease. ZEB1, AR, E-Ca and N-Ca play important roles in prostate cancer progression. ZEB1 can not only affect prostate cancer through epithelial stromal transformation (EMT), but also through AR. ZEB1 may also be related to the development of CRPC.

Expression of POU2F2 in clear cell renal cell carcinoma and its effects on the biological behavior of the cancer cells / 现代泌尿外科杂志

【Objective】 To investigate the expression of transcription factor POU domain class 2 transcription factor 2 (POU2F2) in clear cell renal cell carcinoma (ccRCC) and human renal cancer cell lines (786-O and ACHN) and its effects on the cells’ biological behaviors such as proliferation, migration and invasion in vitro. 【Methods】 The mRNA expressions of POU2F2 in ccRCC tissues, adjacent normal tissues, cell lines 786-O and ACHN were detected with real-time polymerase chain reaction (qRT-PCR). The protein expression of POU2F2 in ccRCC tissues and adjacent normal tissues were detected with immunohistochemistry. The effects of knockdown of POU2F2 on the mRNA and protein expressions of epithelial mesenchymal transformation (EMT)-related tumor markers were detected with qRT-PCR and Western blot. 【Results】 The mRNA expression of POU2F2 in ccRCC tissues was significantly higher than that in adjacent normal tissues, and was correlated with patients’ gender, WHO/ISUP nuclear grade and TNM stage. The protein expression of POU2F2 was significantly higher in ccRCC tissues than in adjacent normal tissues, and was correlated with tumor pathological grade and TNM stage. The mRNA expression of POU2F2 was significantly decreased in 786-O cells after sh-POU2F2-1013 plasmid transfection (P<0.05); the proliferation ability, clonal formation rate, migration ability and invasion ability were significantly reduced (P<0.05). Knockdown of POU2F2 down-regulated the mRNA and protein expressions of MMP2, MMP9 and Twist in 786-O cells, while up-regulated E-ca expression. 【Conclusion】 The mRNA expression of POU2F2 was significantly up-regulated in ccRCC tissues and renal cancer cells. Knockdown of POU2F2 inhibited the proliferation, migration and invasion of cells in vitro, and slowed or inhibited the occurrence and development of renal cancer.

LncRNA UNC5B-AS1 promotes proliferation and epithelial-mesenchymal transition of cervical cancer cells by regulating Toll-like receptor signaling pathway / 中国药理学通报

Aim To investigate the effects of long non-coding RNA(lncRNA)UNC5B-AS1 on the proliferation and epithelial mesenchymal transformation(EMT)of cervical cancer. Methods GEO and TCGA databases were used to download data sets and differential expression analysis was performed. qRT-PCR was used to verify the differential expression of lncRNA UNC5B-AS1 in normal and cancerous cervical tissues.The interference and overexpression of lncRNA UNC5B-AS1 were transfected into cervical cancer cell lines, and plate cloning, CCK-8 and EdU experiments were used to detect the effect of lncRNA UNC5B-AS1 on the pro-liferation of cervical cancer cells.Transwell assay was used to detect its effect on migration and invasion of cervical cancer cells.The expression levels of EMT-related genes E-Cadherin, N-Cadherin and Vimentin were detected by Western blot. Transcriptome sequencing was used to obtain the signal pathway regulated by lncRNA UNC5B-AS1, and to verify the expression level of related genes. Results RNA microarray and bioinformatics analysis showed that the expression level of lncRNA UNC5B-AS1 in cervical cancer was significantly higher than that in normal cervical tissue, and correlated with the overall survival time of patients.Compared with the negative control group, knockdown lncRNA UNC5B-AS1 could reduce the proliferation, migration and invasion of cervical cancer cells, while overexpression could promote the proliferation, migration and invasion of cervical cancer cells. Western blot showed that lncRNA UNC5B-AS1 could regulate EMT of cervical cancer cells. Transcriptome sequencing showed that lncRNA UNC5B-AS1 could regulate Toll like receptor(TLR)signaling pathway. qRT-PCR and Western blot results showed that the expression levels of TLR-related genes IL-6 and TICAM2 in the knockdown and overexpression lncRNA UNC5B-AS1 group were significantly changed(P<0.05). Conclusions LncRNA UNC5B-AS1 is highly expressed in cervical cancer. Overexpression of lncRNA UNC5B-AS1 may enhance TLR signaling pathway activity, thereby promoting proliferation and EMT of cervical cancer cells.

Acteoside Inhibits Epithelial Mesenchymal Transformation of Hepatoma Cells Through Regulation of ERK1/2 Signaling Pathway / 肿瘤防治研究

Objective To investigate the effect and mechanism of acteoside (ACT) in inhibiting epithelial-mesenchymal transition (EMT) in human hepatoma HCCLM3 cells by regulating the ERK1/2 pathway. Methods CCK-8 assay was used to detect the effect of hepatocellular carcinoma cell proliferation. The invasion and migration of HCC cells were detected by scratch and Transwell tests. The mRNA and protein expression levels of the ERK1/2 signaling pathway and EMT-related genes (E-cadherin and N-cadherin) were detected by real-time PCR and Western blot analyses. Results ACT reduced the activity of HCCLM3 cells and inhibited the proliferation of HCC cells, and the effects had certain correlation with drug concentration and time. ACT inhibited the migration and invasion process of HCCLM3 cells in a concentration-dependent manner. ACT downregulated the mRNA and protein expression of genes related to the ERK1/2 signaling pathway. It increased the mRNA and protein expression levels of the EMT-related gene E-cadherin but decreased those of N-cadherin. Conclusion ACT could inhibit EMT and the invasion and migration of HCCLM3 cells in human hepatoma, and the underlying mechanism is closely related to the downregulation of the ERK1/2 signaling pathway.

Advances in the correlation between growth factors and proliferative vitreoretinopathy / 国际眼科杂志(Guoji Yanke Zazhi)

@#Proliferative vitreoretinopathy(PVR)is a common complication of perforation injury and surgery for rhegmatogenous retinal detachment. The pathogenesis of this disease is still unclear. However, studies have shown that retina pigment epithelium(RPE)cells have the ability to secrete cytokines, and many growth factors are overexpressed in vitreous or subretinal fluid in PVR patients. These growth factors and their receptors play an important role in the occurrence and development of PVR. When the blood-retinal barrier is broken, the physiological balance of growth factors disappears, and RPE cells are stimulated by growth factors to undergo epithelial-mesenchymal transformation(EMT), migration and proliferation, this leads to the formation of the preretinal membrane, which pulls on the retina and causes retinal detachment. In recent years, scholars have done a lot of researches on the signaling pathways, EMT process and cell proliferation involved in the formation of PVR with growth factors. This article will summarize the function of growth factors involved in the formation of PVR and the therapeutic effects of antagonistic growth factors in the development of PVR.

Effects of sodium arsenite on liver fibrosis and expression of epithelial-mesenchymal transformation-related proteins in SD rats / 环境与职业医学

Background Long-term exposure to sodium arsenite leads to its accumulation in the liver and liver injury as a result. Previous studies showed that mesenchymal cells play an important role in hepatic fibrosis, and epithelial-mesenchymal transformation (EMT) is considered to be a main source of mesenchymal cells. Objective To investigate the effects of sodium arsenite at different doses on liver fibrosis and EMT-related protein expressions in SD rats. Methods Twenty-four healthy weaned SD rats, half male and half female, were randomly divided into four groups according to body weight, with 6 rats in each group. The four groups were control group (gavage with 10.0 mL·kg−1 physiological saline), 2.5 mg·kg−1 sodium arsenite group, 5.0 mg·kg−1 sodium arsenite group, and 10.0 mg·kg−1 sodium arsenite group. All rats were gavaged 6 d per week for 36 weeks and weighed once a week, the serum and liver tissues of rats were collected and weighed, then the organ coefficient was calculated. Hematoxylin-eosin staining and Masson's trichrome staining were used to determine the pathological changes of hepatic fibrosis in rats. The serum secretion levels of hyaluronic acid (HA), laminin (LN), procollagen Ⅲ N-terminal propeptide (PⅢNP), and collagen Ⅳ (COL-Ⅳ) in rats were detected by enzyme-linked immunosorbent assay (ELISA). The protein expressions of HSCs activation-related proteins, such as α-smooth muscle actin (α-SMA) and transforming growth factor-β1 (TGF-β1), as well as EMT-related markers, such as E-cadherin, N-cadherin, Vimentin, and Snail, were detected by Western blotting. Results Compared with the control group, the 10.0 mg·kg−1 sodium arsenite group showed decreased body weight (P<0.05) and increased liver coefficient (P<0.05) of female and male rats. The pathological staining showed that, compared with the control group, a large number of inflammatory cells were observed in liver tissue of rats exposed to sodium arsenite, liver parenchymal cells were also liquefied, necrotic, and denatured, and the collagen positive staining area of liver tissue showed an upward trend along with the increase of arsenic exposure dose (P<0.05). The results of ELISA and Western blotting showed that the serum secretion levels of HA, LN, PⅢNP, and COL-Ⅳ in the 5.0 and 10.0 mg·kg−1 sodium arsenite groups were higher than those in the control group and the 2.5 mg·kg−1 sodium arsenite group (P<0.05). Compared with the control group, the expressions of α-SMA and TGF-β1 proteins in liver tissue were increased in each sodium arsenite exposure group (P<0.05), the expression levels of E-cadherin protein were decreased (P<0.05), and the expression levels of N-cadherin, Vimentin, and Snail were increased (P<0.05). Conclusion Sodium arsenite exposure can induce HSCs activation and liver fibrosis injury in SD rats, resulting in increased extracellular matrix secretion levels, accompanied by EMT in liver tissue, suggesting that EMT is closely related to the process of liver fibrosis caused by arsenic.