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1.
International Eye Science ; (12): 67-70, 2022.
Article in Chinese | WPRIM | ID: wpr-906732

ABSTRACT

@#Proliferative vitreoretinopathy(PVR)is a eye disease characterized by the formation of epiretinal membranes(ERM)composed of extracellular matrix(ECM)and various types of cells in the vitreous and/or the surface of the retina through the wound repair and fibrotic process. ERM shrinks to form retinal folds and stretches the retina to cause retinal detachment(RD). Epithelial-mesenchymal transition(EMT)of retinal pigment epithelial(RPE)cells and accumulation of ECM are considered to be the main pathological mechanisms for the formation of ERM. RPE cells undergo a process named EMT induced by transforming growth factor-β(TGF-β), by which differentiated epithelial cells go through epithelial phenotypic loss, the weakness of cell-cell contact and mesenchymal phenotype expression. Fibroblast-like cells differentiated from mesenchymal cells produce ECM and other components, which forms ERM together with glial cells and fibroblasts, <i>etc</i>. Recent studies indicated a lot of cytokines/growth factors, transcriptional factors, and microRNA(miRNA)regulate the development of EMT in RPE cells, in which miRNA is a novel and powerful regulatory gene and plays a critical regulatory role in the EMT process of PVR. This review focuses on the current understandings of the mechanism and the interventional treatments of miRNA in PVR.

2.
Arch. med ; 21(1): 24-34, 2021/01/03.
Article in Spanish | LILACS | ID: biblio-1148354

ABSTRACT

Objective: to determine the presence and distribution of markers of the epithelialmesenchymal transition (EMT) (S-100A4 and alpha-smooth muscle actin-α-SMA) in gingival tissues of patients affected by Gingival hypertrophy (GH) due to orthodontics.GH is an exaggerated increase in gingival tissue whose pathogenesis is unknown. However, it has been reported that the epithelial-mesenchymal transition as a process involved in other types of GH. Materials and methods: descriptive study that included the analysis of gingival tissues of healthy individuals (n = 6) and patients with GH by orthodontic treatment (n = 6). Before gingival surgery, the patients underwent a periodontal hygiene phase. The gingival tissue samples obtained were processed and embedded in paraffin. The cuts were made with a microtome and deposited on polysine adhesion slides. Histological hematoxylin-eosin staining was performed.The identification and location of S-100A4 and α-SMA markers was determined by immunohistochemistry with monoclonal antibodies. The reading of the findings was carried out by oral pathologists. Results: in healthy individuals, an S100A4 label was observed in Langerhans cells, while α-SMA was identified in the vascular endothelium of all samples analysed. However, in patients with GH due to orthodontics, they registered an intense staining of S100A4 in gingival fibroblasts, Langerhans cells, vascular endothelium, and areas adjacent to the rupture of blood vessel. α-SMA expression in GO was detected in the vascular endothelium and gingival fibroblasts. Conclusion: the differential immunostaining of EMT markers in gingival tissues of patients with orthodontic GH suggests an eventual role of EMT in the pathogenesis of this pathology..Au


Objective: to determine the presence and distribution of markers of the epithelialmesenchymal transition (EMT) (S-100A4 and alpha-smooth muscle actin-α-SMA) in gingival tissues of patients affected by Gingival hypertrophy (GH) due to orthodontics. GH is an exaggerated increase in gingival tissue whose pathogenesis is unknown. However, it has been reported that the epithelial-mesenchymal transition as a process involved in other types of GH. Materials and methods: descriptive study that included the analysis of gingival tissues of healthy individuals (n = 6) and patients with GH by orthodontic treatment (n = 6). Before gingival surgery, the patients underwent a periodontal hygiene phase. The gingival tissue samples obtained were processed and embedded in paraffin. The cuts were made with a microtome and deposited on polysine adhesion slides. Histological hematoxylin-eosin staining was performed. The identification and location of S-100A4 and α-SMA markers was determined by immunohistochemistry with monoclonal antibodies. The reading of the findings was carried out by oral pathologists. Results: in healthy individuals, an S100A4 label was observed in Langerhans cells, while α-SMA was identified in the vascular endothelium of all samples analysed. However, in patients with GH due to orthodontics, they registered an intense staining of S100A4 in gingival fibroblasts, Langerhans cells, vascular endothelium, and areas adjacent to the rupture of blood vessel. α-SMA expression in GO was detected in the vascular endothelium and gingival fibroblasts. Conclusion: the differential immunostaining of EMT markers in gingival tissues of patients with orthodontic GH suggests an eventual role of EMT in the pathogenesis of this pathology..Au


Subject(s)
Humans , Patients , Tissues , S100 Calcium-Binding Protein A4
3.
Cancer Research and Clinic ; (6): 469-472, 2021.
Article in Chinese | WPRIM | ID: wpr-912907

ABSTRACT

miRNA-27a (miR-27a) is a member of miR-23a-27a-24 gene cluster and is abnormally expressed in gastric cancer. miR-27a plays an important role in gastric cancer cell epithelial-mesenchymal transition, proliferation, microenvironment and resistance to chemotherapy drugs by regulating SFRP1, PHLPP2, BTG2, SOCS6, FOXO1 and other tumor suppressor genes. At the same time, single nucleotide polymorphisms of miR-27a and gastric cancer have also become hotspots in recent years. A more comprehensive exploration of the relationship between miR-27a and gastric cancer is expected to provide new methods and ideas in the treatment of gastric cancer. This article reviews the research progress of miR-27a in gastric cancer.

4.
Chinese Journal of Nephrology ; (12): 911-917, 2021.
Article in Chinese | WPRIM | ID: wpr-911912

ABSTRACT

Objective:To investigate the role and mechanism of (histone deacetylase 6, HDAC6) in the epithelial-mesenchymal transition (EMT) of renal tubular epithelial cells and the activation of renal interstitial fibroblasts.Methods:Human renal tubular epithelial cells (HK-2) and rat renal interstitial fibroblast (NRK-49F) were cultured in vitro, and divided into 4 groups: control group, Tubastatin A (TA) group (treated with 10 μmol/L HDAC6 inhibitor TA for 36 h), transforming growth factor-β1 (TGF-β1) group (10 ng/ml TGF-β1 for 36 h), and TGF-β1+TA group (treated with 10 ng/ml TGF-β1 and 10 μmol/L TA for 36 h). The expression levels of fibronectin, α-smooth muscle actin (α-SMA), collagen I, E-cadherin, HDAC6, acetyl histone H3, histone H3, acetyl α-tubulin, α-tubulin, TGF-β receptor (TGF-βR) 1, p-Smad3, Smad3, connective tissue growth factor (CTGF), epidermal growth factor receptor (EGFR) and p-EGFR in HK-2 and NRK-49F cell samples were detected by Western blotting, and quantitative analysis was performed according to gray level. Results:(1) In HK-2 cells stimulated by TGF-β1, TA decreased the expression of fibronectin, α-SMA, collagen I, and increased the expression of epithelial cell marker E-cadherin. Meanwhile, TA decreased the expression of HDAC6 and increased the expression levels of acetyl histone H3 and acetyl α-tubulin (all P<0.05). (2) Compared with the TGF-β1 group, the expressions of TGF-βR1, p-Smad3, CTGF and p-EGFR in TGF-β1+TA group were decreased (all P<0.05), while the total protein levels of Smad3 and EGFR were not significantly different (both P>0.05). (3) In NRK-49F cells stimulated by TGF-β1, TA decreased the expressions of fibronectin, α-SMA, collagen I, TGF-βR1 and p-Smad3 (all P<0.05). Conclusions:Blockade of HDAC6 by TA may inhibit the EMT of renal tubular epithelial cells and the activation of renal interstitial fibroblasts via regulating multiple signaling pathways including TGF-β/Smad3, CTGF and EGFR.

5.
Chinese Journal of Geriatrics ; (12): 1386-1391, 2021.
Article in Chinese | WPRIM | ID: wpr-911024

ABSTRACT

Objective:To investigate the expression of urinary exosomal miR-150-5p in different age groups and its relationship with kidney aging.Methods:Seventy-six subjects were recruited at the geriatrics department of Beijing Friendship Hospital from January 2014 to December 2016, with 15 in the young group, 35 in the middle-aged group, and 26 in the elderly group.We detected the expression of urinary exosomal miR-150-5p by real-time PCR from urine samples of different age groups and used Logistic regression analysis to evaluate the correlation of different levels of miR-150-5p with age, gender, history of hypertension, history of coronary heart disease, and hyperlipidemia.Additionally after overexpression and knockdown of miR-150-5p in human renal tubular cells of the HK2 cell line, we analyzed the expression levels of epithelial-to-mesenchymal transition(EMT)-related proteins by Western blotting.Results:Logistic regression analysis showed that age was closely related to estimated urinary exosomal miR-150-5p levels( P<0.05), and estimated urinary exosomal miR-150-5p levels in the elderly group(1.33±0.41)were significantly higher than those in the young group(0.88±0.40)( P<0.05). In addition, estimated urinary exosomal miR-150-5p levels became inversely proportional to the glomerular filtration rate as age increased. In vitro experiments showed that the expression levels of ZO-1 and E-cadherin in epithelial cells were slightly decreased after knockdown of miR-150-5p in renal tubular epithelial cells of the HK-2 cell line. Conclusions:The expression of miR-150-5p in urinary exosomes may be slightly increased in the elderly.It could inhibit the occurrence of renal EMT, and its expression is correlated with renal function.Mir-150-5p may also affect the expression of adhesion molecule-related proteins in HK-2 cells.MiR-150-5p in urinary exosomes is expected to become one of the genetic markers associated with renal aging.

6.
Article in Chinese | WPRIM | ID: wpr-910635

ABSTRACT

Objective:To investigate the effect of heme oxygenase-1 (HO-1) modified rat bone marrow mesenchymal stem cells (BMSCs) on fibrotic rats.Methods:110 male SD rats aged 6-8 weeks were selected randomly divided into model group, BMSCs group and HO-1/BMSCs group with 11 rats in each group after intraperitoneal injection of CCl 4, PBS, BMSCs and HO-1/BMSCs were injected respectively. Another 11 rats were selected as control group. After 4 weeks of intervention, tracer experiment was used to detect the location of BMSCs. Rats in each group were executed, and liver function were detected by biochemical analyzer, liver fibrosis indexes were detected by ELISA, liver histopathology were detected by HE and Sirius red staining. Immunohistochemistry, Western blot and RT-PCR were used to detect the protein and mRNA expression of E-cadherin and Vimentin. Results:The rat fibrosis model was successfully established. Tracer experiment showed that BMSCs were implanted in rat liver after transplantation. Compared with the model group, the liver function and liver fibrosis indexes of BMSCs group and HO-1/BMSCs group were improved, and Ishak score and stage were significantly decreased, and HO-1/BMSCs group was superior to BMSCs group. The expression of E-cadherin in HO-1/BMSCs group (0.92±0.21), (0.84±0.03) were higher than those in BMSCs group [(0.54±0.16), (0.53±0.04)] and model group [(0.49±0.06), (0.11±0.06)] both at protein and mRNA level, while protein and mRNA level of Vimentin (1.21±0.23), (3.82±0.80) were lower than that in BMSCs group [(1.32±0.17), (6.39±0.75)] and model group [(1.41±0.18), (16.94±1.30)]. The difference was statistically significant ( P<0.05). Conclusion:HO-1/BMSCs can improve liver function and liver fibrosis in fibrotic rats more effectively than BMSCs alone. The mechanism was possibly through inhibiting liver epithelial mesenchymal transition.

7.
Journal of Chinese Physician ; (12): 842-847, 2021.
Article in Chinese | WPRIM | ID: wpr-909630

ABSTRACT

Objective:To investigate the intervention effect of ventricular zone expressed PH domain-containing 1 (VEPH1) on epithelial mesenchymal transition (EMT) and proliferation of human cutaneous melanoma (CM) cells based onthe transforming growth factor-β (TGF-β) signaling pathway.Methods:The melanoma cells were cultured in vitro. After transfecting the melanoma cells with overexpression or interference plasmids of VEPH1 or TGF-β overexpression plasmids, or treating the cells with SB-431542 (TGF-β pathway inhibitor), we detected the expression of genes and proteins relevant to VEPH1, TGF-β, and EMT by quantitative reverse transcription polymerase chain reaction (qRT-PCR) and Western blot to observe the effect of these proteins on CM cell proliferation. Results:qRT-PCR results showed that the expression of VEPH1 in melanoma cells (B16-BL6, B16 and A375 cells) was significantly lower than that in HaCaT cells, and the lowest expression was found in A375 cells, so A375 cells were selected for follow-up experiments. After transfection with VEPH1 overexpression plasmid or SB-431542, the mRNA and protein expression of E-cadherin in A375 cells were significantly increased, the mRNA and protein expressions of TGF-β, Smad4, N-cadherin and vimentin were significantly decreased, and the cell proliferation was significantly decreased ( P<0.05). Compared with the VEPH1 vector group, the expression of TGF-β, Smad4 and N-cadherin in the VEPH1 vector+ SB-431542 group was significantly reduced ( P<0.05); the expression of E-cadherin was increased, and the cell proliferation was also significantly decreased ( P<0.05). The expression of TGF-β, Smad4, N-cadherin and vimentin were increased after co-transfection with VEPH1 vector, while the expression of E-cadherin was decreased, and the cell proliferation was also enhanced ( P<0.05). The expression of VEPH1 in A375 cells was significantly decreased after transfection with si-VEPH1 plasmid, while that in SB-431542 and TGF-β vector group was not significantly decreased. Conclusions:VEPH1 can inhibit human CM cells by the intervention on TGF-β signaling pathway. This study reveals the potential of VEPH1 as a diagnostic, prognostic and therapeutic target for CM.

8.
Article in Chinese | WPRIM | ID: wpr-907539

ABSTRACT

Objective:To explore the mechanism of a novel BMX splicing variant induced epidermal growth factor receptor tyrosine kinase inhibitor (EGFR-TKI) gefitinib resistance in lung cancer.Methods:Stable transgenic cell line PC9-BMXΔN and HCC827-BMXΔN were constructed by lentivirus infection of PC9 and HCC827 cells carrying EGFR mutation. The cells were divided into PC9-Vec group (PC9 cells transfected with empty vector), PC9-BMX group (PC9 cells stably expressing BMX), PC9-BMXΔN group (PC9 cells stably expressing BMXΔN) and HCC827-Vec group (HCC827 cells transfected with empty vector), HCC827-BMXΔN group (HCC827 cells stably expressing BMXΔN). Quantitative real-time PCR was used to detect the expression levels of mRNA. The protein expression levels in each group were detected by Western blotting. The cells in the PC9-Vec group and PC9-BMXΔN group were treated with 0, 0.01, 2.00, 50.00, 100.00, 200.00 nmol/L and 2.00, 4.00 μmol/L gefitinib. The cells in the HCC827-Vec group and HCC827-BMXΔN group were treated with 0, 0.01, 1.00, 10.00, 100.00 nmol/L and 1.00 μmol/L gefitinib. MTT method was used to detect cell viabilities.Results:The PC9-BMXΔN cells were scattered and showed a fibroblast-like morphology. Compared with the PC9-Vec cells, the relative expression levels of fibronectin, N-cadherin, vimentin, Snail, Slug and TWIST 2 mRNA in PC9-BMXΔN cells were up-regulated. Compared with the PC9-Vec cells and PC9-BMX cells, the expression levels of fibronectin and vimentin protein in PC9-BMXΔN cells were up-regulated; while the expression level of E-cadherin protein in PC9-BMXΔN cells was significantly down-regulated. Compared with the PC9-Vec cells, the cell viabilities of PC9-BMXΔN cells treated with 0.01 nmol/L [(99.11±2.16)% vs. (91.29±1.91)%, t=-4.701, P=0.011], 2.00 nmol/L [(80.41±1.48)% vs. (63.36±2.14)%, t=-11.324, P<0.001], 50.00 nmol/L [(80.83±5.38)% vs. (60.22±3.61)%, t=-5.507, P=0.005], 100.00 nmol/L [(75.54±3.46)% vs. (59.93±1.91)%, t=-6.836, P=0.002], 200.00 nmol/L [(77.57±6.53)% vs. (56.70±2.88)%, t=-5.064, P=0.007], 2.00 μmol/L [(70.22±3.45)% vs. (53.14±0.89)%, t=-8.309, P=0.001], 4.00 μmol/L [(68.66±4.67)% vs. (52.30±2.59)%, t=-4.882, P=0.008] gefitinib were significantly increased, with statistically significant differences. Similarly, compared with the HCC827-Vec cells, the cell viabilities of HCC827-BMXΔN cells treated with 1.00 nmol/L [(64.36±2.49 )% vs. (47.13±4.21)%, t=-7.067, P=0.019], 10.00 nmol/L [(63.25±5.87)% vs. (43.28±2.95)%, t=-5.267, P=0.006], 100.00 nmol/L [(49.47±5.74)% vs. (37.12±4.92)%, t=-2.830, P=0.047], 1.00 μmol/L [(49.05±3.34)% vs. (32.06±4.73)%, t=-5.073, P=0.007] gefitinib were significantly increased, with statistically significant differences. Gefitinib treatment could significantly inhibit the expression levels of p-EGFR and p-ERK1/2 both in PC9-Vec cells, PC9-BMX cells and PC9-BMXΔN cells. Compared with the PC9-Vec cells and PC9-BMX cells, the expression level of p-EGFR in PC9-BMXΔN cells was significantly increased after gefitinib treatment for 8 h (0.91±0.04 vs. 0.81±0.04 vs. 0.80±0.05, all P<0.05); the expression levels of p-ERK1/2 in PC9-BMXΔN cells were significantly increased after gefitinib treatment for 2 h (0.64±0.06 vs. 0.38±0.12 vs. 0.37±0.14), 4 h (1.28±0.06 vs. 1.08±0.06 vs. 1.11±0.07), and 8 h (0.75±0.04 vs. 0.55±0.05 vs. 0.60±0.07), with statistically significant differences (all P<0.05). Conclusion:BMXΔN is involved in EGFR-TKI gefitinib resistance in lung cancer, which may be achieved by inducing cells to undergo epithelial-mesenchymal transition and activating the ERK/MAPK signaling pathway.

9.
Article in Chinese | WPRIM | ID: wpr-907527

ABSTRACT

Objective:To investigate the interaction between heat shock protein 90 (Hsp90) and silent mating-type information regulation 2 homolog 1 (SIRT1) and evaluate its effect on epithelial-mesenchymal transition (EMT) of lung cancer A549 cells.Methods:EMT model was established by treating lung cancer A549 cells with 5 μg/L transforming growth factor-β1 (TGF-β1), which was used as TGF-β1 group, and the normal lung cancer A549 cells were used as control group. The interaction between Hsp90 and SIRT1 in lung cancer A549 cells was detected by immunocoprecipitation method. The expression of Hsp90 gene was silenced by RNA interference technique, and the cells were divided into TGF-β1 group, TGF-β1+ siRNA-Hsp90-neg group and TGF-β1+ siRNA-Hsp90 group. Transwell invasion assay was used to investigate the effect of the interaction of Hsp90 and SIRT1 on the invasion ability of lung cancer A549 cells. The expressions of Hsp90, SIRT1, E-cadherin and vimentin were detected by Western blotting. The effect of inhibiting Hsp90 expression on the stability of SIRT1 protein and EMT of lung cancer A549 cells was observed.Results:After 48 h induction with TGF-β1, EMT characteristics of lung cancer A549 cells were induced successfully. The relative expression levels of Hsp90 protein in the control group and TGF-β1 group were 0.45±0.05 and 1.31±0.06, respectively, the relative expression levels of SIRT1 protein were 0.29±0.04 and 0.95±0.08, respectively, and there were statistically signigicant differences ( t=10.98, P=0.018; t=7.39, P=0.028). The results of immunocoprecipitation showed that there was an interaction between Hsp90 and SIRT1 protein in lung cancer A549 cells. The relative expression levels of Hsp90 in the TGF-β1 group, TGF-β1+ siRNA-Hsp90-neg group and TGF-β1+ siRNA-Hsp90 group were 0.75±0.07, 0.63±0.06 and 0.23±0.05, respectively, and there was a statistically significant difference ( F=18.85, P=0.012). The relative expression levels of SIRT1 in the above three groups were 0.99±0.08, 0.97±0.12 and 0.35±0.05, respectively, and there was a statistically significant difference ( F=16.52, P=0.014). The expression levels of Hsp90 and SIRT1 in the TGF-β1+ siRNA-Hsp90 group were significantly lower than those in the TGF-β1 group ( P=0.019, P=0.016). The numbers of cells passing Matrigel in the above three groups were 378.13±27.70, 323.52±19.82 and 142.51±22.54, respectively, and there was a statistically significant difference ( F=27.35, P=0.022). The number of cells passing Matrigel in the TGF-β1+ siRNA-Hsp90 group was significantly less than that in the TGF-β1 group ( P=0.028). The relative expression levels of E-cadherin in the above three groups were 0.31±0.02, 0.34±0.04 and 0.63±0.05, respectively, and there was a statistically significant difference ( F=19.39, P=0.031). The relative expression levels of vimentin in the above three groups were 0.33±0.02, 0.27±0.05 and 0.09±0.03, respectively, and there was a statistically significant difference ( F=12.58, P=0.012). The expression level of E-cadherin in the TGF-β1+ siRNA-Hsp90 group was significantly higher than that in the TGF-β1 group ( P=0.017), while the expression level of vimentin was significantly lower than that in the TGF-β1 group ( P=0.023). Conclusion:Hsp90 interacts with SIRT1, and Hsp90 inhibition can lead to the decrease of SIRT1 protein level. Hsp90 may play a role of molecular chaperone to maintain the conformation stability of SIRT1, and the interaction between Hsp90 and SIRT1 may be one of the molecular mechanisms for the occurrence of EMT and the enhancement of invasion ability of lung cancer A549 cells.

10.
International Journal of Surgery ; (12): 401-405, 2021.
Article in Chinese | WPRIM | ID: wpr-907451

ABSTRACT

CCL20 and CCR6 are chemokines produced by a variety of cells. CCL20 and CCR6 combine to stimulate a series of downstream pathways, participate in the occurrence and development of various malignant tumors, and also play an important role in the invasion and metastasis of breast cancer and the process of chemotherapy resistance. Epithelial-mesenchymal transformation (EMT) is a key step in the process of tumor cell metastasis, which is characterized by loss of cell adhesion, down-regulation of E-cherherin expression, up-regulation of mesenchymal markers and fibrinectin expression, and enhancement of cell motor ability and invasion ability. This article reviews the research of CCL20-CCR6 biological axis and EMT on invasion and metastasis of breast cancer cells.

11.
Article in Chinese | WPRIM | ID: wpr-906330

ABSTRACT

Objective:To investigate the effects of artesunate (ART) on epithelial-mesenchymal transformation (EMT) of colorectal cancer HCT-8 cells,and explore the effects of ART on cell migration,invasion,EMT ability, and protein kinase B (Akt)/Snail signaling pathway of colorectal cancer. Method:3-(4-5-Dimethylthiazol-2-yl)2,5-diphenyltetrazolium bromide (MTT) assay was used to detect the effects of ART at different concentrations on the proliferation of HCT-8 cells. Wound healing assay and Transwell assay were used respectively to detect the effects of ART on migration and invasion of colorectal cancer cells. The effects of different concentrations of ART on the distribution of EMT-related proteins vimentin and E-cadherin in HCT-8 cells were detected by double-immunofluorescent staining. The effects of ART on protein expression levels of EMT markers E-cadherin,vimentin and N-cadherin in HCT-8 cells and the expression of Akt1, p-Akt1, and Snail1 in the Akt/Snail signaling pathway were determined by Western blot. Result:The dose-dependent inhibitory effects of ART on the proliferation of HCT-8 cells were determined and the inhibition rate was calculated. A dose-response curve was plotted accordingly. The half-maximal inhibitory concentration (IC<sub>50</sub>) of ART on HCT-8 cells was (16.67±1.95) μmol·L<sup>-1</sup>. The following four groups were set up: a control group (0 μmol·L<sup>-1</sup>),and low-, medium-, and high-dose ART groups(2, 10, 50 μmol·L<sup>-1</sup>). Compared with the results in the control group,ART inhibited the migration and invasion of HCT-8 cells(<italic>P</italic><0.05). Specifically, the expression of E-cadherin in HCT-8 cells was significantly up-regulated,and that of vimentin and N-cadherin was significantly down-regulated (<italic>P</italic><0.05). The expression levels of p-Akt1 and Snail1 were significantly decreased after ART treatment,thus inhibiting EMT(<italic>P</italic><0.05). Conclusion:The findings of this study suggested that ART inhibited the EMT-triggered migration and invasion of HCT-8 cells presumedly by inhibiting the activation of the Akt/Snail pathway to reverse EMT.

12.
Article in Chinese | WPRIM | ID: wpr-906175

ABSTRACT

Objective:To study the effect of Fuzheng Qufeng prescription (FZQP) on transforming growth factor-<italic>β</italic><sub>1</sub> (TGF-<italic>β</italic><sub>1</sub>)/Smad signaling pathway and epithelial-mesenchymal transition of podocyte in membranous nephropathy (MN) rats and to explore its molecular mechanism for podocyte protection. Method:The rats were randomly divided into normal control group (NC) and modeling group. Rats in modeling group induced by bovine serum albumin (C-BSA) were randomly divided into model group (MN), losartan potassium group (LP, 0.05g·kg<sup>-1</sup>), and FZQP high dose (FZQPH, 41 g·kg<sup>-1</sup>), medium dose (FZQPM, 20.5 g·kg<sup>-1</sup>), and low dose (FZQPL, 10.25 g·kg<sup>-1</sup>) groups. The administration lasted for 4 weeks. In week 0, 2, and 4 of administration, the levels of 24 hours urine protein (24 h-Upro) were tested. At the end of 4th week, the levels of blood urea nitrogen (BUN) and serum creatinine (SCr) were detected, and the rats in each group were sacrificed and the renal pathological morphology changes were observed by light microscope with hematoxylin-eosin (HE), Masson and periodic acid-silver metheramine (PASM) staining. The deposition of immune complex, the thickening of glomerular basement membrane (GBM) and podocyte foot process were observed by transmission electron microscope (TEM). The distribution and expression intensity of Desmin in renal tissues were detected by immunohistochemistry (IHC). The mRNA and protein expression levels of TGF-<italic>β</italic><sub>1</sub>, Smad2/3, phospho(p)-Smad2/3, Smad7 and Desmin in renal tissues were respectively detected by Western blot (WB) and Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR). Result:Compared with NC group, the levels of 24 h-Upro, BUN and SCr significantly increased in model group (<italic>P</italic><0.01), with increased deposition of immune complex, significantly thickened GBM and fusion of foot processes, significantly increased Desmin mRNA and protein expression (<italic>P</italic><0.01) and increased TGF-<italic>β</italic><sub>1</sub>, Smad2, and Smad3 mRNA and protein expression (<italic>P</italic><0.05), and decreased Smad7 mRNA and protein expression (<italic>P</italic><0.05,<italic>P</italic><0.01). Compared with model group, 24 h-Upro and BUN decreased in FZQP groups and LP group (<italic>P</italic><0.05), levels of serum SCr in FZQPM group decreased (<italic>P</italic><0.05), deposition of immune complex, thickening of GBM and fusion of foot process were all alleviated in FZQP groups and LP group. Distribution of Desmin along GBM decreased in FZQPH group, FZQPM group and LP group (<italic>P</italic><0.05). Both mRNA and protein expression levels of TGF-<italic>β</italic><sub>1</sub> and p-Smad2/Smad2 in FZQPM group decreased, while mRNA and protein expression levels of Smad7 increased (<italic>P</italic><0.05). Both mRNA and protein expression levels of p-Smad3/Smad3 in FZQPH group decreased (<italic>P</italic><0.05). Both mRNA and protein expression levels of Desmin in podocyte in FZQPH group, FZQPM group and LP group decreased (<italic>P</italic><0.05). Conclusion:FZQP might realize podocyte protection effect in MN via suppressing EMT mediated by overactivated TGF-<italic>β</italic><sub>1</sub>/Smad signaling pathway.

13.
Article in Chinese | WPRIM | ID: wpr-905954

ABSTRACT

Objective:To study the efficacy and mechanism of Zishenwan (ZSW) against pyroptosis and epithelial-mesenchymal transition (EMT) of renal tubular epithelial cells in diabetic nephropathy (DN) mice, so as to provide evidence for the treatment of DN with ZSW. Method:The <italic>db/db</italic> mice with spontaneous diabetes were randomly divided into the model group, dapagliflozin (1.0 mg·kg<sup>-1</sup>) group, and high-, medium-, and low-dose (6.0, 3.0, 1.5 g·kg<sup>-1</sup>) ZSW groups. The non-diabetic <italic>db/m</italic> mice were classified into the normal group. The ones in the model and normal groups were given an equal volume of deionized water by gavage, while those in the other groups were intervened with the corresponding drugs for 12 weeks. The fasting blood glucose (FBG) level was tested at tail vein once every two weeks. The levels of urine albumin-creatinine ratio (ACR), <italic>β</italic>-N-acetyl-D-glucosaminidase (NAG), and cystatin C (CysC) were detected once every four weeks. After 12 weeks of administration, the blood sampled from eyeballs was used for measuring the blood urea nitrogen (BUN) and serum creatinine (SCr). The pathological changes in renal tissues were observed by light microscopy and transmission electron microscopy. The expression of EMT markers in the renal tubular epithelium was analyzed by immunohistochemistry (IHC). The in situ terminal end-labeling (TUNEL) staining was conducted to analyze the nuclear damage of renal tubular epithelial cells. The protein and mRNA expression levels of EMT markers, nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) inflammasome and pyroptosis-related inflammatory cytokines in renal tissues were separately assayed by Western blot and Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR). Result:Compared with the normal group, the model group displayed significantly increased FBG, BUN, serum SCr, ACR, NAG, and CysC (<italic>P</italic><0.01), impaired renal tissues, altered EMT marker expression intensities and levels (<italic>P</italic><0.01), and elevated TUNEL-positive rate and protein and mRNA expression levels of pyroptosis-related inflammatory cytokines and NLRP3 inflammasome (<italic>P</italic><0.01). Compared with the model group, ZSW and dapagliflozin significantly decreased the levels of FBG, BUN, serum SCr, ACR, NAG, and CysC (<italic>P</italic><0.01), relieved the pathological injuries in renal tissues, changed the EMT marker expression intensities (<italic>P</italic><0.01) and protein and mRNA expression levels (<italic>P</italic><0.05, <italic>P</italic><0.01), and down-regulated the TUNEL-positive rate (<italic>P</italic><0.01) of renal tubular epithelial cells as well as the protein and mRNA expression levels of pyroptosis-related inflammatory cytokines (<italic>P</italic><0.01) and NLRP3 inflammasome (<italic>P</italic><0.05, <italic>P</italic><0.01). Conclusion:ZSW alleviates DN possibly by inhibiting pyroptosis and EMT in renal tubular epithelial cells.

14.
Article in Chinese | WPRIM | ID: wpr-905953

ABSTRACT

Objective:To investigate the effect of Banxia Xiexintang on the epithelial-mesenchymal transition (EMT) of human peritoneal mesothelial cell line (HMrSV5) induced by gastric cancer-derived exosomes (Exo). Method:Banxia Xiexintang-containing serum was prepared and the human gastric cancer NCI-N87-derived exosomes (NCI-N87-Exo) were extracted, followed by their identification by transmission electron microscopy and Western blotting and labeling with 1,1-dioctadecyl-3,3,3,3- tetramethylindocarbocyanine perchlorate (Dil). The cells were divided into the blank group, model group, and low-, medium-, and high-dose (13.5,27,54 g·kg<sup>-1</sup>) Banxia Xiexintang groups. HMrSV5 cells in the blank group were cultured alone, the ones in the model group with 100 mg·L<sup>-1</sup> NCI-N87-Exo, and those in the low-, medium-, and high-dose Banxia Xiexintang groups with 100 mg·L<sup>-1</sup> NCI-N87-Exo plus low-, medium-, and high-dose 10% Banxia Xiexintang-containing serum, respectively. Confocal laser microscope was used to observe the uptake of NCI-N87-Exo by HMrSV5 cells at 24 h, 48 h and 72 h. Seventy-two hours later, the morphological changes in HMrSV5 cells were observed. The protein expression levels of E-cadherin, cytokeratin 19 (CK19), <italic>α</italic>-smooth muscle actin (<italic>α</italic>-SMA), elastin, and transforming growth factor-<italic>β</italic><sub>1</sub> (TGF-<italic>β</italic><sub>1</sub>), Smad2/3, and p-Smad2/3 were assayed by Western blot. Result:It was observed under the transmission electron microscope that NCI-N87-Exo showed an oval or dish-shaped vesicle structure with a particle size ranging from 40 to 80 nm. Exo marker proteins CD9 and CD63 were highly expressed while calreticulin was not expressed, implying that the NCI-N87-Exo was confirmed. After 24 h, 48 h, 72 h of co-culture, it was observed under the fluorescence microscope that NCI-N87-Exo were taken up by HMrSV5 cells, which was positively correlated with time. Compared with the blank group, Banxia Xiexintang significantly inhibited the uptake of NCI-N87-Exo by HMrSV5 cells, with better effect noticed in the middle- and high-dose Banxia Xiexintang groups(<italic>P</italic><0.05,<italic>P</italic><0.01). After intervention with Banxia Xiexintang-containing serum, the HMrSV5 cells were arranged densely, and the intercellular space was significantly reduced, with the most obvious changes present in the high-dose Banxia Xiexintang group. Western blot revealed that the protein expression levels of E-cadherin and CK19 in HMrSV5 cells after being intervened with the medium- and high-dose Banxia Xiexintang-containing serum were increased significantly as compared with those in the blank group, whereas the levels of <italic>α</italic>-SMA and Elastin were decreased significantly (<italic>P</italic><0.01). Banxia Xiexintang-containing serum at the low, medium, and high doses remarkably down-regulated TGF-<italic>β</italic><sub>1</sub> and p-Smad2/3 protein expression(<italic>P</italic><0.05,<italic>P</italic><0.01). However, there was no significant change in Smad2/3. Conclusion:NCI-N87-Exo can be taken up by HMrSV5 cells to induce EMT. Banxia Xiexintang can inhibit the uptake of NCI-N87-Exo by HMrSV5 cells and the resulting EMT induced by NCI-N87-Exo, which is related to the regulation of TGF-<italic>β</italic><sub>1</sub>/Smads signaling pathway.

15.
Article in Chinese | WPRIM | ID: wpr-905902

ABSTRACT

Objective:To observe the effect of oxymatrine (OM) combined with bevacizumab ( BV ) on the proliferation, invasion, and migration of breast cancer MCF-7 cells and explore the mechanism of OM in regulating BV-induced epithelial-mesenchymal transition (EMT) based on the Wnt/<italic>β</italic>-catenin signaling pathway. Method:The effect of different concentrations of OM(0, 0.5, 1.0, 2.0, 4.0, 8.0, 16.0 mmol·L<sup>-1</sup>)and BV(0, 0.25×10<sup>-4</sup>, 0.50×10<sup>-4</sup>, 1.00×10<sup>-4</sup>, 2.00×10<sup>-4</sup>, 4.00×10<sup>-4</sup>, and 8.00×10<sup>-4</sup> mmol·L<sup>-1</sup>)on the proliferation of MCF-7 cells were detected by cell counting kit-8(CCK-8)assay. The effect of OM(4.0 mmol·L<sup>-1</sup>) combined with BV(2.00×10<sup>-4</sup> mmol·L<sup>-1</sup>)on the invasion and migration of MCF-7 cells were observed in transwell and scratch repair tests. Western blot was conducted to investigate the effect of OM(4.0 mmol·L<sup>-1</sup>)combined with BV (2.00×10<sup>-4</sup> mmol·L<sup>-1</sup>) on proliferation-related proteins in MCF-7 cells, followed by the detection of the expression levels of Wnt/<italic>β</italic>-catenin signaling pathway- and EMT-related proteins. Result:Compared with the blank group, OM (2.0,4.0,8.0,16.0 mmol·L<sup>-1</sup>) inhibited the proliferation of MCF-7 cells in a concentration-dependent manner (<italic>P</italic><0.01), while BV did not show the inhibitory effect against the proliferation of MCF-7 cells. The inhibitory effect of the combination of the two drugs on the proliferation of MCF-7 cells was not significantly different from that of OM. Compared with the blank group, OM significantly reduced the migration distance of MCF-7 cells and the number of invaded cells(<italic>P</italic><0.01), while BV increased the migration distance of MCF-7 cells and the number of invaded cells (<italic>P</italic><0.05,<italic>P</italic><0.01). Compared with BV, its combination with OM significantly inhibited the invasion and migration of MCF-7 cells induced by BV (<italic>P</italic><0.01). Compared with the blank group, both OM and the combined medication obviously inhibited the phosphorylation of proliferation-related protein kinase B(Akt) and extracellular-signal-regulated protein kinase 1/2 (ERK1/2)in MCF-7 cells (<italic>P</italic><0.01) and down-regulated the protein expression levels of <italic>β</italic>-catenin, proto-oncogene (c-Myc), CD44, and G<sub>1</sub>/S-specific cyclin D<sub>1</sub> in Wnt/<italic>β</italic>-catenin signaling pathway (<italic>P</italic><0.05,<italic>P</italic><0.01). Besides, OM and the combination of two drugs both significantly reduced the protein expression levels of calcium-dependent cell adhesion protein <italic>N</italic>-cadherin and Vimentin in EMT, whereas increased the expression of calcium-dependent cell adhesion protein E-cadherin(<italic>P</italic><0.01). However, the expression of the above-mentioned proteins in the BV group was reversed (<italic>P</italic><0.05,<italic>P</italic><0.01). Conclusion:After the combination with BV, OM plays an anti-breast cancer role by effectively inhibiting the activation of Wnt/<italic>β</italic>-catenin pathway induced by BV and reversing EMT.

16.
Article in Chinese | WPRIM | ID: wpr-905862

ABSTRACT

Objective:To investigate the effect of Astragalus polysaccharide (APS) on transforming growth factor-<italic>β</italic><sub>1</sub> (TGF-<italic>β</italic><sub>1</sub>)-induced epithelial mesenchymal transition (EMT) of A549/DDP lung adenocarcinoma xenograft and its potential molecular mechanism. Method:BALB/c nude mice were randomly divided into the non-loading group (A549/DDP cells not loaded with TGF-<italic>β</italic><sub>1</sub>), model group, cisplatin group, and combined group (A549/DDP cells overexpressing TGF-<italic>β</italic><sub>1</sub>). Mice in the combined group were treated with intragastric administration of APS (0.3 g·kg<sup>-1</sup>·d<sup>-1</sup>) and intraperitoneal injection of cisplatin (0.003 5 g·kg<sup>-1</sup>), while those in the cisplatin group only received intraperitoneal injection of cisplatin (0.003 5 g·kg<sup>-1</sup>). After drug intervention, the nude mice were sacrificed and the xenograft and lung were harvested, followed by the weighing of tumor and the calculation of the inhibition rate. The number of tumors metastasizing to the lung was counted under the microscope. The pathological features of tumors and their metastasis to the lung tumor were observed by hematoxylin-eosin (HE) staining. The protein and mRNA expression levels of EMT molecular markers E-cadherin, Vimentin, <italic>α</italic>-smooth muscle actin (<italic>α</italic>-SMA), and phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt) in the xenograft were detected by immunohistochemistry, Western blot, and Real-time polymerase chain reaction (Real-time PCR). Result:Compared with the non-loading group, the model group exhibited increased tumor weight and pulmonary metastatic nodules (<italic>P</italic><0.05), sparse tumor cell junctions, long spindle cells, massive metastatic nodules in the lung, down-regulated E-cadherin protein and mRNA expression, and up-regulated Vimentin and <italic>α</italic>-SMA protein and mRNA expression and p-PI3K and p-Akt protein expression (<italic>P</italic><0.05). Compared with the model group and cisplatin group, the combined group displayed decreased tumor weight and pulmonary metastatic nodules (<italic>P<</italic>0.05), tight tumor cell junctions, round or oval cells, no obvious lung metastasis, up-regulated E-cadherin protein and mRNA expression (<italic>P</italic><0.05), and down-regulated Vimentin and <italic>α</italic>-SMA protein and mRNA expression (<italic>P</italic><0.05) and p-PI3K and p-Akt protein expression (<italic>P</italic><0.05). There was no significant difference in PI3K or Akt protein expression among groups. Conclusion:APS has a certain inhibitory effect against EMT in lung adenocarcinoma A549/DDP cells, which may be related to the inhibition of activated PI3K/Akt protein expression.

17.
Article in Chinese | WPRIM | ID: wpr-905830

ABSTRACT

Objective:This studu aims to investigate the effect of aqueous extract of modified Xiao Xianxiongtang on the epithelial mesenchymal transition(EMT) and the change of its invasion and migration ability of human gastric cancer MGC-803 cells mediated by transforming growth factor-<italic>β</italic><sub>1</sub>(TGF-<italic>β</italic><sub>1</sub>),and to explore the mechanism of regulating Wnt5a/Ca<sup>2+</sup>/ activated T-cell nuclear factor(NFAT) signaling pathway to inhibit EMT and invasion and metastasis of MGC-803 cells. Method:TGF-<italic>β</italic><sub>1</sub>(10 μg·L<sup>-1</sup>)was used to induce EMT and the invasion and metastasis model of human gastric cancer MGC-803 cells. Transwell chamber experiment, scratchhealing experiment, Western blot and immunofluorescence assay were used to detect cell invasion and migration ability, expression of EMT marker protein and key protein expression of Wnt5a/Ca<sup>2+</sup>/NFAT signaling pathway, and intracellular Ca<sup>2+</sup> concentration. Result:Compared with the blank group, TGF-<italic>β</italic><sub>1</sub> could significantly enhance the invasion and migration ability of MGC-803 cells(<italic>P</italic><0.01), down-regulate the level of E-cadherin(<italic>P</italic><0.01), up-regulate protein expressions of N-cadherin, Snail and Vimentin(<italic>P</italic><0.01), and induce cell Wnt5a, calcineurin (CaN), total protein of activated T-cell nuclear factor 1(NFAT1), up-regulation of phosphorylated proteins p-NFAT1 and NFAT1 nucleoprotein and intracellular accumulation of Ca<sup>2+</sup>(<italic>P</italic><0.01). Compared with the TGF-<italic>β</italic><sub>1</sub> group, modified Xiao Xianxiongtang (10, 20, 40 mg·L<sup>-1</sup>) could significantly inhibit this phenomenon,and 40 mg·L<sup>-1</sup> had the best effect(<italic>P</italic><0.05,<italic>P</italic><0.01).The specific inhibitors of Wnt5a/Ca<sup>2+</sup>/NFAT signaling pathway (<italic>R</italic>)-(+)-Bay-K-8644 and modified Xiao Xianxiongtang (40 mg·L<sup>-1</sup>) could significantly inhibit theinvasion and migration of MGC-803 cells mediated by TGF-<italic>β</italic><sub>1</sub>, up-regulate the level of E-cadherin, and down-regulate expressions of N-cadherin, Snail, Vimentin, Wnt5a, CaN and NFAT1 proteins and reduce the intracellular accumulation of Ca<sup>2+</sup>(<italic>P</italic><0.05,<italic>P</italic><0.01).Moreover, (<italic>R</italic>)-(+)-Bay-K-8644 combined with modified Xiao Xianxiongtang (40 mg·L<sup>-1</sup>) had stronger inhibitory effect(<italic>P</italic><0.05,<italic>P</italic><0.01). Conclusion:These results suggest that modified Xiao Xianxiongtang can inhibit the EMT mediated by TGF-<italic>β</italic><sub>1</sub> via Wnt5a/Ca<sup>2+</sup>/NFAT signaling pathway,thereby reducing the invasion and migration ability of MGC-803 cells.

18.
International Eye Science ; (12): 2104-2108, 2021.
Article in Chinese | WPRIM | ID: wpr-904683

ABSTRACT

@#Proliferative vitreoretinopathy(PVR)is a serious complication arisen from ocular trauma, diabetic retinopathy, vascular retinopathy, inflammatory retinopathy and other ocular diseases. It is also the most important reason for the failure of rhegmatogenous retinal detachment surgery, which is a great threat of visual function. A large number of studies have proved that the main risk factor for PVR is the damage of blood-retinal barrier, in which retinal pigment epithelial(RPE)cells are stimulated by cytokines in the vitreous cavity. RPE cells underwent epithelial-mesenchymal transition(EMT), which transformed into fibroblasts. The cell morphology changed, the tight junctions between cells disappeared, the cell polarity lost, and the proliferation, migration, and invasion abilities were enhanced. A contractile fibrous proliferative membrane is formed on the anterior surface or under the retina. The fibrous proliferative membrane will lead to the retina folds, pull the retina and lead to retinal detachment, which will eventually lead to vision loss or even blindness. Nowadays, plenty of studies investigating the prevention and treatment of PVR have been carried out at home and abroad. In this review, we briefly illustrated the signaling pathways related to epithelial-mesenchymal transformation in RPE cells and the treatment of PVR.

19.
Journal of Medical Biomechanics ; (6): E659-E663, 2021.
Article in Chinese | WPRIM | ID: wpr-904452

ABSTRACT

In the process of tumor growth, with the proliferation and expansion of cancer cells, the reconstruction of extracellular matrix (ECM) of cancer tissues, the restriction of surrounding tissues and the flow of cancer tissue interstitial fluid, the special stress environment is formed in the tumor tissues. Significant differences are found in the mechanical environment and mechanical characteristics of different regions of tumor tissues, that is, mechanical heterogeneity. The reseach shows that the mechanical properties of tumor tissue invasion frontier areas are more significant and complex. In particular, the epithelial-mesenchymal transition (EMT) of tumor cells also prefers to concentrate on this area. The mechanical stress generated by the invasion front can induce EMT of tumor cells through TWIST1, TGF-β, WNT and other force signal transduction pathways, and promote tumor cell invasion. From the perspective of tumor biomechanics, this review focuses on the relationship between mechanical heterogeneity of tumor cells and EMT, so as to provide the theoretical basis for mechanoenvironment-targeted therapy of tumors.

20.
Acta Pharmaceutica Sinica B ; (6): 1274-1285, 2021.
Article in English | WPRIM | ID: wpr-881198

ABSTRACT

Liver is the most common metastatic site for colorectal cancer (CRC), there is no satisfied approach to treat CRC liver metastasis (CRCLM). Here, we investigated the role of a polycomb protein BMI-1 in CRCLM. Immunohistochemical analysis showed that BMI-1 expression in liver metastases was upregulated and associated with T4 stage, invasion depth and right-sided primary tumor. Knockdown

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