Background: Hepatic fibrosis is attributed to an imbalance of extracellular matrix production and lysis. Human hepatic stellate cells (HSCs) have been uncovered to converge through complex interactions with hepatocytes and immune cells, causing scarring in liver damage. Aims: We aimed to investigate the expression status of ubiquitin specific peptidase 1 (USP1) and its potential mechanisms on HSCs and hepatic fibrosis . Methods: Hepatic fibrosis animal and cell models were generated using mice with carbon tetrachloride (CCl4) treatment and HSCs LX-2 with TGF- beta 1 treatment. Relationships among USP1, SNAIL, and CXCL1 were identified via dual-luciferase reporter gene assay, co-immunoprecipitation, and chromatin immunoprecipitation. With gain- and loss-of-experiments, CCK-8 and flow cytometry assays were employed for cell proliferation and apoptosis. Results: USP1 upregulated SNAIL expression through deubiquitination to increase CXCL1 expression. USP1 downregulation decreased expressions of fibrosis-related genes, suppressed proliferation, and promoted apoptosis in TGF- beta 1-induced LX-2 cells, which were reversed by SNAIL overexpression. The profibrosis role caused by SNAIL upregulation was abolished by CXCL1 reduction. Promotive function of USP1/SNAIL/CXCL1 axis in hepatic fibrosis was further confirmed in vivo . Conclusion: These data supported siRNA-mediated silencing of USP1 improved hepatic fibrosis through inhibition of SNAIL and CXCL1, which yields a new therapeutic target for hepatic fibrosis treatment. (c) 2021 Editrice Gastroenterologica Italiana S.r.l. Published by Elsevier Ltd. All rights reserved.

Hepatic fibrosis (HF) is involved in aggravated wound-healing response as chronic liver injury. Extracellular vesicles (EVs) carrying microRNA (miR) have been reported as therapeutic targets for liver diseases. In this study, we set out to explore whether adipose-derived mesenchymal stem cells (ADMSCs)-derived EVs containing miR-150-5p affect the progression of HF. Carbon tetrachloride (CCl4 ) was firstly used to induce HF mouse models in C57BL/6J mice, and activation of hepatic stellate cells (HSCs) was achieved using transforming growth factor β (TGF-β). EVs were then isolated from ADMSCs and co-cultured with HSCs. The relationship between miR-150-5p and CXCL1 was identified using dual luciferase gene reporter assay. Following loss- and gain-function experimentation, HSC proliferation was examined by MTT assay, and levels of fibrosis-, HSC activation- and apoptosis-related genes were determined in vitro. Additionally, pathological scores, collagen volume fraction (CVF) as well as levels of inflammation- and hepatic injury-associated genes were determined in in vivo. Down-regulated miR-150-5p and elevated CXCL1 expression levels were detected in HF tissues. ADMSCs-derived EVs transferred miR-150-5p to HSCs. CXCL1 was further verified as the downstream target gene of miR-150-5p. Moreover, ADMSCs-EVs containing miR-150-5p markedly inhibited HSC proliferation and activation in vitro. Meanwhile, in vivo experiments also concurred with the aforementioned results as demonstrated by inhibited CVF, reduced inflammatory factor levels and hepatic injury-associated indicators. Both experiments results were could be reversed by CXCL1 over-expression. Collectively, our findings indicate that ADMSCs-derived EVs containing miR-150-5p attenuate HF by inhibiting the CXCL1 expression.

Background: Mesenchymal stem cell (MSC) therapy can prevent parenchymal cell loss and promotes tissue repair through the action of trophic, secreted molecules. In this study, we investigated whether MSC-conditioned medium (MSC-CM) could protect hepatocytes and sinusoidal endothelial cells (SECs) and stimulate their regeneration in 50% reduced-size liver transplantation (RSLT).Materials and methods: Rats were randomly divided into three groups: sham-operated group, MSC-CM group (rats with 50% RSLT receiving MSC-CM infusion), and medium group (rats with 50% RSLT receiving medium therapy). Graft function, proinflammatory cytokines, incidence of apoptosis, proliferation of hepatocytes and SECs, and the expression of vascular endothelial growth factor and matrix metallopeptidase 9 were assessed in this study.Results: Systemic infusion of MSC-CM prevented the release of liver injury biomarkers and provided a significant survival benefit. Furthermore, MSC-CM therapy resulted in reduction of apoptosis of hepatocytes and SECs. The number of proliferating hepatocytes and SECs increased 1.2- and 1.6-fold, respectively, accompanied by a decrease in the expression levels of several proinflammatory cytokines and a noticeable decrease in infiltration of neutrophils and activation of Kupffer cells. Also, increased expression of vascular endothelial growth factor and matrix metallopeptidase 9 in the grafts was observed after MSC-CM therapy.Conclusions: These data suggest that MSC-CM therapy in RSLT provides trophic support to the injured liver by inhibiting SEC and hepatocellular death and stimulating their regeneration. (c) 2013 Elsevier Inc. All rights reserved.

Mesenchymal stem cell (MSC) therapy can prevent hepatic parenchymal cell loss and promote tissue repair. However, poor MSC engraftment is one of the primary barriers to the effectiveness of cell therapy because culture-expanded MSCs progressively down-regulate C-X-C chemokine receptor type 4 (CXCR4) expression and lose their ability to migrate toward a concentration gradient of stromal cellderived factor 1a (SDF1a). In this study, we investigated whether a CXCR4-MSC infusion could protect hepatocytes and stimulate regeneration in 50% reduced size liver transplantation (RSLT). Rats that underwent 50% RSLT were randomly divided into 3 groups: a phosphate-buffered solution group (PBS), a green fluorescent protein (GFP)MSC group, and a CXCR4-MSC group. Rats received 1 mL of PBS with or without a resuspension of GFP-MSCs or CXCR4-MSCs. The factors secreted by MSCs, the graft function, the apoptosis and proliferation of hepatocytes, the efficacy of MSC engraftment, and the expression of SDF1, albumin (Alb), and cytokeratin 18 (CK18) in engrafted GFP-positive MSCs were assessed. A systemic infusion of GFP-MSCs led to a reduction of the release of liver injury biomarkers and apoptosis of hepatocytes; CXCR4 overexpression did not further reduce the liver injury. However, CXCR4 overexpression enhanced MSC engraftment in liver grafts, improved the effect on the proliferation of hepatocytes, and thus provided a significant 1-week survival benefit. SDF1 expression in grafts was elevated after transplanted CXCR4-MSCs were recruited to the remnant liver. However, engrafted MSCs did not express the markers of hepatocytes, including Alb and CK18, in vivo 168 hours after transplantation. CXCR4 overexpression enhanced the mobilization and engraftment of MSCs into small-for-size liver grafts, in which these cells promoted the early regeneration of the remnant liver not by direct differentiation but perhaps by a paracrine mechanism. Liver Transpl 19:215-225, 2013. (c) 2012 AASLD.

Tumor resistance to chemoradiation therapy is partly attributed to the presence of apoptosis-resistant cancer stem cells (CSCs). Chemoradiation therapy can enrich CSCs by killing apoptosis-susceptible cancer cells.

Objective To investigate the significance of pancreatic cancer cells resistance to chemoradiotherapy induces EMT.Methods Several pancreatic cancer cell lines were exposed to gemzar while radiotherapy was given at the same time.After about two weeks,when 90% pancertic cancer cells were killed,a small number of cells resistant to chemoradiotherapy was obtained,and their morphological changes were observed by microscopy;transwell test was used to detect invasiveness,and western-blot was used to detect expression of epithelial marker E-cad and mesenchymal marker vimentin.Results Pancreatic cancer cells resistance to chemoradiotherapy showed phenotypic changes consistent with EMT: spindle-cell shape,loss of polarity,and pseudopodia formation.BxPC3,jf305 and pc3 cells resistance to chemoradiotherapy exhibited an increase in migrating and invading(P﹤0.05,respectively).Pancreatic cancer cells resistance to chemoradiotherapy also had decreased expression of E-cadherin and an increase in vimentin.Conclusions Pancreatic cancer cells resistance to chemoradiotherapy can induce epithelial-mesenchymal transition(EMT),and EMT may be related to resistance of chemoradiotherapy of pancreatic cancer.

Objective To investigate the expression of Bcl-2,survivin and pancreatic cancer stem cells markers Oct-4 and ABCG2 in pancreatic cancer cells resistance to chemoradiotherapy.and explore its mechanism.Methods Concurrent ehemoradiotherapy was used to obtain pancreatic cancer cells resistant to chemoradiotherapy,the pancreatic cancer cells without chemoradiotherapy treatment were used as control.Western-blot was applied to detect the expression of Bcl-2,survivin,Oct-4,ABCG2.Results The expression of Bcl-2 was 0.7955±0.0326,0.5718±0.0212,0.6137±0.0382 and 0.8733±0.0461,respectively;the expression of survivin protein was 0.8207±0.0490,0.6973±0.0211,0.7967±0.0346 and 0.8013±0.0398,respectively;the expression of Oct-4 protein was 0.8728±0.0177,0.7861±0.0139,0.4794±0.0932 and 0.4216±0.1043,respectively;the expression of ABCG2 protein was 0.7810±0.1370,0.4957±0.1126,0.6102±0.1358 and 0.4670±0.1274,respectively.in resistant pancreatic cancer cells of SW1990,BxPC3,pc3,jf305 cell line.The corresponding values in the control group were 0.4723±0.018,0.2954±0.0103.0.3587±0.0201 and 0.2718±0.0136;0.4717±0.0274,0.3587±0.0113,0.3891±0.0147 and 0.3326±0.0124;0.6053±0.0142,0.4236±0.0086.0.2385±0.0671 and 0.1985±0.0582;0.3156±0.0582.0.2360±0.0423,0.2813±0.0512 and 0.1808±0.a0370.The expression of all the four proteins significantly increased after ehemoradiotherapy(P＜0.05).Conclusions Pancreatic cancer cells resistant to chemoradiotherapy may contain cancer stem cells.

目的 分析胚胎干细胞标志物Oct-4在胰腺癌组织和胰腺癌细胞株中的表达状况.方法 应用RT-PCR检测Oct-4 mRNA在25例胰腺癌组织以及胰腺癌细胞株SW1990、PC3、JF305和BxPC3中的表达;应用Western blot法检测胰腺癌组织及胰腺癌细胞株中Oct-4蛋白表达;应用异硫氰荧光素 (FITC)标记的流式细胞仪分选法检测Oct-4在胰腺癌细胞株中的表达率.结果 4株胰腺癌细胞株均表达Oct-4 mRNA和Oct-4 蛋白.88%(22/25)胰腺癌组织高表达Oct-4 mRNA和Oct-4 蛋白,其表达率与胰腺癌的分化程度无关.Oct-4在SW1990的表达率为50.21%、BxPC3为68.53%、PC3为66.27%、JF305为67.61%.结论 Oct-4异常表达可能与胰腺癌始发有关,胰腺癌可能来源于胰腺干细胞.