Xu T , Pan L , Li L , Hu S , Zhou H , Yang C , Yang J , Li H , Liu Y , Meng X , Li J .

1607a0202062 The fund of Anhui Science and Technology Department Soft Science Project, 81700522 National Natural Science Foundation of China, 1704a0802161 Anhui Provincial Natural Science Foundation, 1808085MH235 Anhui Provincial Natural Science Foundation, 0601067101 the Fund of Anhui Medical University Doctoral Start Research, 2018xkj021 Anhui Medical University Scientific Research Foundation

	Figure 2. TMEM88 was decreased in TGF‐β1‐stimulated LX‐2 cells. A, The kinetic profiles of TMEM88 expression were observed in LX‐2 cells during activation induced by TGF‐β1 in different times and concentrations. B, The slides of LX‐2 cells transfected with pEGFP‐C2‐TMEM88 were taken out to immunofluorescence, and the result showed that the TMEM88 protein was mainly localized in cytoplasm. C, D, RT‐qPCR and Western blotting detected the expression level of TMEM88 both at mRNA and protein levels when transfected with pEGFP‐C2‐TMEM88 and pEGFP‐C2, respectively (GFP fluorescence protein‐positive cells were sorted after 48 h using MoFlo XDP cell Sorter of Center For Scientific Research, Beckman Coulter). The results were expressed as the mean ± standard of three different experiments. *P < .05 compared with the control group, # P < .05 compared with the control group
	Figure 3. TMEM88 inhibited HSCs activation in TGF‐β1‐stimulated LX‐2 cells. A, The mRNA expression levels of α‐SMA and Co1.I were analysed with RT‐qPCR in activated LX‐2 cells transfected with pEGFP‐C2‐TMEM88 and TMEM88‐siRNA, respectively. The results showed that TMEM88 could inhibit the mRNA expression levels of α‐SMA and Co1.I. B, The protein expression levels of α‐SMA and Co1.I were measured by Western blotting analysis in activated LX‐2 cells transfected with pEGFP‐C2‐TMEM88 and TMEM88‐siRNA. The results showed that TMEM88 could inhibit the protein expression levels of α‐SMA and Co1.I. The results were expressed as the mean ± standard of three different experiments. *P < .05 compared with the normal group, # P < .05 compared with the control group
	Figure 4. TMEM88 alleviated ECM accumulation in TGF‐β1‐stimulated LX‐2 cells. A, The mRNA expression levels of MMP2 and TIMP1 were analysed with RT‐qPCR in activated LX‐2 cells transfected with pEGFP‐C2‐TMEM88 and TMEM88‐siRNA, respectively. The results showed that TMEM88 could decrease the mRNA expression level of TIMP1, whereas increase the mRNA expression level of MMP2. B, The protein expression levels of MMP2 and TIMP1 were measured by Western blotting analysis in activated LX‐2 cells transfected with pEGFP‐C2‐TMEM88 and TMEM88‐siRNA. The results showed that TMEM88 could decrease the protein expression level of TIMP1, whereas increase the protein expression level of MMP2. The results were expressed as the mean ± standard of three different experiments. *P < .05 compared with the normal group, # P < .05 compared with the control group
	Figure 5. TMEM88 inhibited cell proliferation and promoted cell apoptosis in TGF‐β1‐stimulated LX‐2 cells. A, Cell apoptosis of LX‐2 cells was measured by flow cytometry analysis. B, Proliferation of LX‐2 cells was determined by EDU DNA incorporation assay. Fluorescence microscope was used for the imaging. C, The TMEM88‐3′‐UTR constructs or blank plasmid were transfected into LX‐2 cells with control or miR‐708 mimics, followed by dual‐luciferase assays. D, The mutant 3′UTR‐TMEM88 constructs or blank plasmid were transfected into LX‐2 cells with control or miR‐708 mimics, followed by dual‐luciferase assays. E, The mRNA expression level of miR‐708 was analysed with RT‐qPCR in human fibrotic liver tissues and normal tissues. F, The mRNA expression level of miR‐708 was analysed with RT‐qPCR in TGF‐β1‐stimulated LX‐2 cells. The results were expressed as the mean ± standard of three different experiments. *P < .05 compared with the normal group, # P < .05 compared with the control group
	Figure 6. MiR‐708 promoted cell activation by targeting TMEM88 in TGF‐β1‐stimulated LX‐2 cells. A, The mRNA and protein expression level of TMEM88 was measured by Western blotting and RT‐qPCR analysis respectively in activated LX‐2 cells transfected with miR‐708 mimics and miR‐708 inhibitor, respectively. B, The mRNA expression levels of α‐SMA and Co1.I were analysed with RT‐qPCR in activated LX‐2 cells transfected with miR‐708 mimics and miR‐708 inhibitor, respectively. The results showed that miR‐708 could increase the mRNA expression levels of α‐SMA and Co1.I. C, The protein expression levels of α‐SMA and Co1.I were measured by Western blotting analysis in activated LX‐2 cells transfected with miR‐708 mimics and miR‐708 inhibitor, respectively. The results showed that miR‐708 could increase the protein expression levels of α‐SMA and Co1.I. The results were expressed as the mean ± standard of three different experiments. *P < .05 compared with the normal group, # P < .05 compared with the control group
	Figure 7. MiR‐708 enhanced ECM accumulation on Wnt/β‐catenin signalling pathway in TGF‐β1‐stimulated LX‐2 cells. A, The mRNA expression levels of MMP2 and TIMP1 were analysed with RT‐qPCR in activated LX‐2 cells transfected with miR‐708 mimics and miR‐708 inhibitor, respectively. The results showed that miR‐708 could increase the mRNA expression level of TIMP1, whereas decrease the mRNA expression level of MMP2. B, The protein expression levels of MMP2 and TIMP1 were measured by Western blotting analysis in activated LX‐2 cells transfected with miR‐708 mimics and miR‐708 inhibitor, respectively. The results were expressed as the mean ± SD of three different experiments. The results showed that miR‐708 could increase the protein expression level of TIMP1, whereas decrease the protein expression level of MMP2. C, The protein expression level of β‐catenin, Wnt3a and Wnt2b was performed in activated LX‐2 cells transfected with pEGFP‐C2‐TMEM88 and TMEM88‐siRNA, respectively. The results were expressed as the mean ± standard of three different experiments. *P < .05 compared with the control group, # P < .05 compared with the control group

Abdel-Al, miRNA-221 and miRNA-222 are promising biomarkers for progression of liver fibrosis in HCV Egyptian patients. 2018, Pubmed

Abdel-Al, miRNA-221 and miRNA-222 are promising biomarkers for progression of liver fibrosis in HCV Egyptian patients. 2018, Pubmed
Adamovich, AMPK couples p73 with p53 in cell fate decision. 2014, Pubmed
Adan, Cell Proliferation and Cytotoxicity Assays. 2016, Pubmed
Affo, The Role of Cancer-Associated Fibroblasts and Fibrosis in Liver Cancer. 2017, Pubmed
Ahmadzadeh, Wnt/β-catenin signaling in bone marrow niche. 2016, Pubmed
Cai, Transmembrane protein 88 attenuates liver fibrosis by promoting apoptosis and reversion of activated hepatic stellate cells. 2016, Pubmed
Campana, Regression of Liver Fibrosis. 2017, Pubmed
Caviglia, MicroRNA-21 and Dicer are dispensable for hepatic stellate cell activation and the development of liver fibrosis. 2018, Pubmed
Chen, TGF-β1 Induces EMT in Bovine Mammary Epithelial Cells Through the TGFβ1/Smad Signaling Pathway. 2017, Pubmed
Czaja, Hepatic inflammation and progressive liver fibrosis in chronic liver disease. 2014, Pubmed
de Leon, Transmembrane protein 88 (TMEM88) promoter hypomethylation is associated with platinum resistance in ovarian cancer. 2016, Pubmed
Freeman, Brain-derived neurotrophic factor and airway fibrosis in asthma. 2017, Pubmed
Gong, Protective effect of miR-20a against hypoxia/reoxygenation treatment on cardiomyocytes cell viability and cell apoptosis by targeting TLR4 and inhibiting p38 MAPK/JNK signaling. 2019, Pubmed
Hatori, Osteoblasts and osteocytes express MMP2 and -8 and TIMP1, -2, and -3 along with extracellular matrix molecules during appositional bone formation. 2004, Pubmed
Hemmann, Expression of MMPs and TIMPs in liver fibrosis - a systematic review with special emphasis on anti-fibrotic strategies. 2007, Pubmed
Hernández-Aquino, Naringenin prevents experimental liver fibrosis by blocking TGFβ-Smad3 and JNK-Smad3 pathways. 2017, Pubmed
Hyun, MicroRNAs in liver fibrosis: Focusing on the interaction with hedgehog signaling. 2016, Pubmed
Jimenez-Mateos, Role of MicroRNAs in innate neuroprotection mechanisms due to preconditioning of the brain. 2015, Pubmed
Koyama, Liver inflammation and fibrosis. 2017, Pubmed
Lee, Mechanisms of hepatic fibrogenesis. 2011, Pubmed
Lee, Identification of transmembrane protein 88 (TMEM88) as a dishevelled-binding protein. 2010, Pubmed , Xenbase
Lee, TMEM88 Inhibits Wnt Signaling by Promoting Wnt Signalosome Localization to Multivesicular Bodies. 2019, Pubmed
Li, MicroRNA-708 is downregulated in hepatocellular carcinoma and suppresses tumor invasion and migration. 2015, Pubmed
Liu, Myofibroblast-specific YY1 promotes liver fibrosis. 2019, Pubmed
Manka, Fibrosis in Chronic Liver Disease: An Update on Diagnostic and Treatment Modalities. 2019, Pubmed
Perumal, Morin attenuates diethylnitrosamine-induced rat liver fibrosis and hepatic stellate cell activation by co-ordinated regulation of Hippo/Yap and TGF-β1/Smad signaling. 2017, Pubmed
Ponsuksili, Genetic architecture and regulatory impact on hepatic microRNA expression linked to immune and metabolic traits. 2017, Pubmed
Prestigiacomo, Pro-fibrotic compounds induce stellate cell activation, ECM-remodelling and Nrf2 activation in a human 3D-multicellular model of liver fibrosis. 2017, Pubmed
Prideaux, MMP and TIMP temporal gene expression during osteocytogenesis. 2015, Pubmed
Rong, Human bone marrow mesenchymal stem cells-derived exosomes alleviate liver fibrosis through the Wnt/β-catenin pathway. 2019, Pubmed
Sun, HBOA ameliorates CCl4-incuded liver fibrosis through inhibiting TGF-β1/Smads, NF-κB and ERK signaling pathways. 2019, Pubmed
Sun, Relevance function of microRNA-708 in the pathogenesis of cancer. 2019, Pubmed
Tokunaga, Selective inhibitor of Wnt/β-catenin/CBP signaling ameliorates hepatitis C virus-induced liver fibrosis in mouse model. 2017, Pubmed
Wen, Interactions between Th1 cells and Tregs affect regulation of hepatic fibrosis in biliary atresia through the IFN-γ/STAT1 pathway. 2017, Pubmed
Wu, Hepatic stellate cells: a target for the treatment of liver fibrosis. 2000, Pubmed
Xu, MicroRNA-708 modulates Hepatic Stellate Cells activation and enhances extracellular matrix accumulation via direct targeting TMEM88. 2020, Pubmed , Xenbase
Xu, TMEM88 mediates inflammatory cytokines secretion by regulating JNK/P38 and canonical Wnt/β-catenin signaling pathway in LX-2 cells. 2018, Pubmed , Xenbase
Yu, Cytosolic TMEM88 promotes triple-negative breast cancer by interacting with Dvl. 2015, Pubmed
Yu, Loss of lncRNA-SNHG7 Promotes the Suppression of Hepatic Stellate Cell Activation via miR-378a-3p and DVL2. 2019, Pubmed
Yu, MicroRNA-17-5p-activated Wnt/β-catenin pathway contributes to the progression of liver fibrosis. 2016, Pubmed
Zhang, Disrupting the TRIB3-SQSTM1 interaction reduces liver fibrosis by restoring autophagy and suppressing exosome-mediated HSC activation. 2020, Pubmed
Zhao, TMEM88 inhibits extracellular matrix expression in keloid fibroblasts. 2017, Pubmed
Zhou, Pathogenesis of liver cirrhosis. 2014, Pubmed