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2022 ◽  
Vol 12 (3) ◽  
pp. 653-658
Author(s):  
Xin Yang ◽  
Shandan Wang

This study intends to promote bone marrow mesenchymal stem cells (BMSCs) differentiation into neural stem cells by down-regulating p38 MAPK/NF-κB to heal neurodegeneration. 26 patients with neurodegenerative diseases were enrolled from the Department of Neurology along with recruitment of 26 other healthy controls followed by analysis of p38 MAPK/NF-κB signaling pathway expression by ELISA. BMSCs were cultured and characterized by flow cytometry. Western blot and qRTPCR measured the p38 MAPK/NF-κB expression in the absence or presence of p38 MAPK/NF-κB inhibitors. p38 MAPK/NF-κB expression in 26 neurodegenerative patients was significantly higher than that of 26 healthy controls. The qRT-PCR and western blot results showed that the neural stem cell-specific proteins expression was increased as days went; after addition of p38 MAPK/NF-κB inhibitor, the expression of related specific genes were significantly decreased. In conclusion, inhibition of the expression of p38 MAPK/NF-κB signaling pathway can heal neurodegeneration by promoting the differentiation of BMSCs into neural stem cells.


2022 ◽  
Vol 12 (3) ◽  
pp. 647-652
Author(s):  
Yang Zhang ◽  
Peng Sun ◽  
Shuying Han ◽  
Duojiao Fan

Mitochondrial autophagy and inflammatory response involves in diabetes. This study mainly explores the role of Silent Information Regulator (SIRT1) in pancreatic β-cells under high glucose conditions and related mechanism. Pancreatic β cells was cultured in a high-glucose environment with SRT1720 and EX527 respectively to define activation group and inhibition group followed by analysis of SIRT1, P-FOXO1, FOXO1, LC3, ATG5, PINK, Parkin, Mfn1, Mfn2, Fis1, IL-6, TNF-α, NLRP3 protein and mRNA expression by qRT-PCR, Western blot and fluorescent probe technology. Compared with control group, SIRT1 protein and mRNA expression in the high glucose group was significantly reduced. Activation group had highest protein and mRNA expression of SIRT1 P-FOXO1, FOXO1, Mfn1, Mfn2, Fis1, PINK, Parkin and mitochondrial membrane potential followed by blank group and inhibition group.SIRT1 secretion by pancreatic β-cells under high glucose environment is reduced. After activating SIRT1, mitochondrial autophagy decreased significantly and inflammatory response is significantly alleviated, indicating that SIRT1 might be used as a therapeutic target.


2022 ◽  
Vol 12 (3) ◽  
pp. 558-563
Author(s):  
Boxian Zhao ◽  
Weiguo Zhu

Multiple miRNAs are differentially expressed in gastric cancer (GC). Herein, this study aims to investigate miR-455’s role in GC and its mechanism. Exosomes (exo) separated from BMSCs after transfection were co-cultured with either phagocytes, GC cells (NCI-N87 cell), or macrophages combined with NCI-N87cells (mixed group) followed by analysis of the expression of PTEN, N-cadherin, E-cadherin, and PI3K, and AKT by RT-qPCR and Western blot. Increased miR-455 expression was observed in GC cells upon transfection. GC cells in the mixed group relative to NCI-N87 group exhibited a lower cell migration and invasion and impaired proliferative capacity (p < 0.05), accompanied with higher expressions of N-cadherin, E-cadherin, PI3K, and AKT, and decreased level of PTEN (p < 0.05). The combined treatment resulted in a higher phagocytic rate (12.38±0.21%) and phagocytic index (14.29±2.11%) compared to treatment with only phagocytes (p < 0.05). In conclusion, BMSC-derived exosomal miR-455 inhibits the growth of GC cells and promotes the phagocytosis through inactivating PI3K/AKT signaling pathway.


2022 ◽  
Vol 12 (3) ◽  
pp. 659-664
Author(s):  
Wei Li ◽  
Tieying Shan ◽  
Jianping Shi ◽  
Zexian Fu ◽  
Shujing Qi ◽  
...  

Extracted MenSC (Menstrual blood-derived stem cells) from female menstrual blood. Added various exogenous factors in-vitro and simulated the female uterine environment to observe how to make MenSC differentiation into Endometrial epithelial cells by artificial induction. MenSCs were divided into 4 groups: 2.5×10−5 mol/L E group, 1.613 nmol/L EGF group, 2.5×10−5 mol/L E+1.613 nmol/L EGF group, control Group (only MenSCs); the relevant indicators of the experiment includes cell staining and Western Blot to detect CK and VIM protein content; RT-PCR to detect CK-19 mRNA and VIM mRNA. The cell staining results showed that E+EGF group had significant differentiation in 7 days and 14 days. CK-19mRNA of E+EGF group was significantly higher than other groups, and the EGF group expression was obviously higher than that of E group, and VIMmRNA expression is opposite to that. The protein expression had the similar performance. MenSC can differentiate into endometrial epithelial cells after induced by E and EFG; and the co-culture of E and EFG can achieve better differentiation, which proves their work together in MenSC differentiate towards endometrial epithelial cells.


2022 ◽  
Vol 12 (4) ◽  
pp. 867-872
Author(s):  
Qunwei You ◽  
Wenjie Wang ◽  
Taotao Tao ◽  
Tianyu Wang ◽  
Danhong Zhang ◽  
...  

This study intends to explore miR-129’s effect on cell viability of Alzheimer’s disease by regulating the target gene APP. The hippocampal neurons were assigned into model group (MO group); mimetic group (SI group); inhibitor group (IN group) followed by analysis of hippocampal neuronal cell proliferation and activity, APP protein content, miR-129 expression and cell apoptosis by CCK-8 assay, Western blot method, MTT assay, qRT-PCR and flow cytometry. miR-129 expression of hippocampal neurons in IN group was lowest. Compared with IN and MO groups, SI group had significantly increased miR-129 level and reduced number of hippocampal neuron apoptosis (P < 0.05). Compared with IN group, MO group had significantly reduced cell apoptosis (P < 0.05). SI group had highest number of hippocampal neurons proliferation followed by IN group. SI group had highest OD value followed by MO group and IN group. The cell activity of SI group was higher than that of IN group and MO group (both P < 0.05). Compared with SI group, rat neuron activity in MO group was significantly higher than IN group (P < 0.05). The APP protein expression of hippocampal neuron cells in SI group was lowest followed by MO group and IN group (P < 0.05). In conclusion, the low miR-129 expression can inhibit the activity of hippocampal neurons possibly through up-regulation of APP protein content.


2022 ◽  
Vol 12 (4) ◽  
pp. 800-806
Author(s):  
Jing Cao ◽  
Fan Yang ◽  
Haiyan Zhou ◽  
Duojiao Fan ◽  
Hengzhou Li ◽  
...  

Our study explores whether BMSC-exosomes overexpressing miR-141 can regulate Wnt signal to inhibit the malignant biological behavior of glioma cells. Thirty healthy mice were selected to construct a glioma mouse model and assigned randomly into the control group, miR-141 NC group, and miR-141 mimic group followed by analysis of cell proliferation, apoptosis, protein expression and mRNA expression by MTT method, flow cytometry, Western blot and RT-PCR methods. Compared with the other two groups, miR-141 mimic group showed reduced number of cell proliferation at 24 h and 48 h, decreased cell migration and invasion ability, and the increased cell apoptosis rate (P < 0.05). In miR-141 mimic group, the protein expression of miR-141 was the highest, while the protein expression of β-catenin, survivin and c-myc was the lowest (P < 0.05). In conclusion, BMSC-exosomes overexpressing miR-141 can inhibit the malignant biological behavior of GC cells possibly by inhibiting the activation of Wnt signaling pathway.


2022 ◽  
Vol 12 (5) ◽  
pp. 1040-1045
Author(s):  
Jingfang Zhu ◽  
Jianglin Hu

Preeclampsia (PE) causes serious harm to the health of mothers and infants. PTEN regulates cell biological behaviors, but its role in preeclampsia have not been reported. Real time PCR and Western blot detected PTEN level in the placenta of PE patients and controls. Placental trophoblastderived cell line HTR8 was assigned into NC group, PTEN group and si-PTEN inhibitor group followed by measuring PTEN level, cell proliferation by MTT assay, cell invasion by Transwell, Caspase 3 activity, Beclin-1 and Atg-5 expression as well as PI3K/Akt/HIF-1α/VEGF signaling protein by Western blot. PTEN in PE patients was significantly downregulated (P < 0.05). Transfection of PTEN siRNA significantly down-regulated PTEN, promoted cell proliferation and invasion, reduced Caspase 3 activity, increased Beclin-1 and Atg-5, and PI3K/Akt/HIF-1α/VEGF protein expression (P < 0.05). Transfection of pcDNA 3.0-PTEN up-regulated PTEN and significantly reversed the above changes (P < 0.05). In conclusion, PTEN is reduced in PE and it can regulate pre-eclampsia trophoblast autophagy possibly through PI3K/Akt/HIF-1α/VEGF signaling, suggesting that PTEN can be a potential target for PE therapy.


2022 ◽  
Vol 12 (5) ◽  
pp. 907-913
Author(s):  
Liyan Zhong ◽  
Yi Yi ◽  
Qian Liu ◽  
Yan Peng

This study intends to discuss the mechanism of MTH1 inhibitor (TH588) in the biological activity of ovarian carcinoma cells. A2780 and SKOV-3 cells were treated with different concentrations of TH588 and assigned into AT group (control), BT group (8 μmol/L TH588), CT group (16 μmol/L), DT group (32 μmol/L), ET group (64 μmol/L) and FT group (128 μmol/L) followed by measuring level of Bcl-2 and Bax by Western blot and PCR, and cell biological activities by MTT, FCM and Transwell chamber assay. The cell proliferative rate was not affected in AT group, but was lower in other groups in a reverse dose-dependent manner. There was significant difference on apoptotic rate and cell invasion among groups with increased apoptosis and reduce invasion after TH588 treatment. FT group showed lowest expression of Bcl-2 and Bax compared to other groups. In conclusion, the biological activity of A2780/SKOV3 cells could be reduced by MTH1 inhibitor which was probably through regulation of Bax and Bcl-2 expression.


2022 ◽  
Vol 12 (5) ◽  
pp. 914-919
Author(s):  
Yilidana Mijiti ◽  
Fang Fang ◽  
Shanhui Liang ◽  
Xiuju Huang ◽  
Yilidana Yilihamu ◽  
...  

The miRNA derived from Bone marrow mesenchymal stem cells (BMSCs) have crucial effects on tumors. The tumor could be affected by the abnormal expression of miRNA in human papillomavirus (HPV). Our study aimed to identify the potential brand-new biomarker in order to reveal the pathogenesis of HPV. miRNA derived from BMSCs was detected and identified. The action of miR-12 on biological behavior of HPV was detected. The level of AN1 protein was detected by Western-blot and IHC method. The relationship between miR-12 and AN1 was assessed by bioinformatics analysis and luciferase assay. The tumor cell biological behaviors were evaluated by manipulating miR12 and AN1 level. The tumor volume derived from BMSCs was diminished significantly compared with normal tissues. The tumor volume was bigger after combined injection with Hela cell and miR-12 compared with single injection. The cell proliferative and invasive ability was strengthened after transfection with miR-12mimics. The cell invasive ability was reduced significantly after transfection of si-miR-12. AN1 was a target gene of miR-12 as confirmed by the analysis on bioinformatics and luciferase activity. The phenotype was reversed after the silent presentation of AN1 was disturbed. In conclusion, miR-12 expression is elevated in HPV cells and affects HPV cells through targeting the AN1 signaling pathway.


2022 ◽  
Vol 12 (5) ◽  
pp. 926-932
Author(s):  
Xin Guan ◽  
Ning Sun

High expression of E74-like factor 3 (ELF3) has been reported in type 1 endometrial cancer (EC). Bioinformatics analysis predicted a positive correlation with ELF3 and mucin 1 (MUC1)/hypoxiainducible factor 1α (HIF-1α), a previously identified cancer-promoting pathway. This study focused on the MUC1/HIF-1α-involved action mechanism of ELF3 in EC. ELF3 expression in EC cell lines was measured by RT-qPCR and western blot analysis. Following the expression of ELF3 was silent, cell proliferation was examined using CCK-8 and colony formation assay, cell migration and invasion were observed using wound healing and transwell assays. The effect of ELF1 silencing on MUC1/HIF-1α expression was detected by western blot. Rescue experiments incorporating pcDNA3.1(+)/MUC1 explored the interaction between ELF3 and MUC1/HIF-1α in EC cell proliferation, migration and invasion. ELF3 was found to be expressed at a high level in EC cell lines, and the silencing of it effectively inhibited EC cell proliferation. Moreover, ELF silencing also inhibited the migration and invasion of EC cells. Consistent with the database prediction, a positive correlation between ELF3 and MUC1/HIF-1α was observed. More importantly, MUC1 overexpression abated the promotive effect of ELF3 silencing on EC cell proliferation, migration and invasion. ELF3 promotes EC cell proliferation, migration and invasion by regulating MUC1/HIF-1α pathway. Thus, ELF3 as well as MUC1/HIF-1α pathway may be particle targets in the treatment of EC.


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