Journal of Translational Medicine
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2026
·Vol. 24
·期 1
·492
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IF 9.7
Li Ze,Ling Yimin,Xiao Wei,Li JiaZheng,Rong JiaHui,Zhang Zewen,Wei Yan,Huo Zhongchao,Li Dong
Background Scutellarin (SCU) is a natural flavonoid compound exhibiting anti-tumor activity. This study aimed to elucidate the molecular mechanism underlying SCU’s therapeutic effects in liver cancer. Methods An integrated approach combining network pharmacology and transcriptomic analysis was employed to identify SCU-related targets, followed by the identification of core targets through protein–protein interaction (PPI) network construction and topological analysis using MCC and MCODE algorithms. A circular RNA (circRNA)-mediated competing endogenous RNA (ceRNA) regulatory network was constructed by integrating data from databases including miRWALK and circBANK with whole-transcriptome sequencing. Molecular interactions were experimentally validated using Western blotting, dual-luciferase reporter assays, and RNA immunoprecipitation (RIP). In Huh7 and HepG2 cells, overexpression and knockdown models of key RNAs were established to evaluate their functional roles, with cell proliferation, cell cycle distribution, and apoptosis assessed via CCK-8, colony formation, and flow cytometry assays. To investigate in vivo anti-tumor efficacy and associated mechanisms, a subcutaneous xenograft tumor model was established in nude mice. Results Integrated network pharmacology and transcriptomic analysis identified 11 core genes significantly enriched in the estrogen signaling pathway, among which ESR1 was validated as a key target. SCU downregulated ESR1 expression in a dose-dependent manner. Mechanistically, SCU upregulated miR-2682-5p while downregulating circSRBD1. Dual-luciferase reporter assays confirmed that miR-2682-5p directly binds to the 3′UTRs of both circSRBD1 and ESR1, supporting the existence of a circSRBD1/miR-2682-5p/ESR1 ceRNA regulatory axis. Functional studies demonstrated that overexpression of circSRBD1 attenuated SCU-induced cell cycle arrest, apoptosis, and suppression of proliferation, whereas silencing circSRBD1 enhanced these effects. Furthermore, SCU and fulvestrant exhibited synergistic anti-tumor activity both in vitro and in vivo. Conclusion This study identifies a novel circSRBD1/miR-2682-5p/ESR1 ceRNA regulatory axis that plays a critical role in mediating SCU’s inhibitory effects on liver cancer progression, highlighting the potential therapeutic value of combining SCU with fulvestrant. Clinical trial number Not applicable.
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EXCLI Journal
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2026
·
IF 6.2
Yunxiao Lin,Fan Yang,Yingtian Zhang,Xiaoqin Zhang,Xiangyi Li,Zhiliang Chen,Xianglong Zhao,Yongzhi Wang,Hao Wu,Cong Huai,Qiange Xiao,Wei Bao,Minglei Yang,Ruoyu Chen,Zhongyu Cao,Jinhang Zhu,Zekun Yu,Zexiu Zhang,Shengying Qin
Liver cirrhosis represents a terminal stage of chronic liver disease with limited pharmacological options. Pien Tze Huang (PTH), a traditional Chinese medicine with established hepatoprotective properties, has shown therapeutic potential in liver fibrosis, yet its molecular mechanisms in cirrhosis remain poorly understood. Here, we investigated the anti-cirrhotic effects of PTH and underlying mechanisms through whole-transcriptome sequencing in a carbon tetrachloride (CCl 4 )-induced mouse model of cirrhosis. PTH treatment significantly attenuated liver injury and other cirrhotic phenotypes, as evidenced by reduced serum transaminases and improved histological features. Transcriptomic analysis identified 1,518 genes dysregulated in cirrhosis and reversed by PTH treatment, with functional enrichment implicating inflammatory and fibrotic pathways including NF-κB, TNF, and IL-17 signaling. Connectivity Map analysis revealed similarity between PTH-induced transcriptional signatures and those of statins and beta-blockers, drugs currently evaluated in cirrhosis clinical trials. Through integrative construction of competing endogenous RNA (ceRNA) networks, we identified a core regulatory module comprising 6 miRNAs, 8 mRNAs, and 27 circRNAs with reversed expression patterns following PTH treatment. Notably, the liver-specific miR-122-5p exhibited the highest network connectivity, targeting P4ha1 , a rate-limiting enzyme in collagen biosynthesis. Human proteome data confirmed liver-enriched expression of P4HA1 , which is a validated therapeutic target of Lufironil, a Phase II clinical trial drug for cirrhosis. Our findings suggest that PTH ameliorates cirrhosis through multi-pathway regulation of a P4ha1 -centered ceRNA network. Functional validation in hepatocytes confirmed that PTH modulates this network via the miR-122-5p/ P4HA1 axis, alleviating injury-induced miR-122-5p downregulation and consequent P4HA1 upregulation. These results provide experimental evidence for a mechanistic basis of PTH's therapeutic application and highlight P4ha1 as a promising biomarker for cirrhosis intervention. See also the graphical abstract (Fig. 1) .
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PATHOLOGY RESEARCH AND PRACTICE
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2021
·
IF 3.25
Yang Yang,Guoying Zhang,Jian Li,Rui Gong,Yingbao Wang,Yang Qin,Qinrong Ping,Libing Hu
Background Increasing evidences have proved that long noncoding RNAs (lncRNAs) regulate the occurrence of bladder cancer (BC) and participate in various pathophysiology processes. However, little is unknown about the role of lncRNAs in drug resistance of BC cells. In this study, we explored the role of non-coding RNA activated by DNA damage (NORAD) in the gemcitabine (GEM) resistant of BC cells and explored its potential mechanism. Methods Real-time quantitative PCR (RT-qPCR) was used to detect the expression of NORAD and miR-155–5p of BC cells. Cell counting kit-8 (CCK-8) and Western blot were used to detect cell inhibition rate and the expression of WEE1 G2 checkpoint kinase (WEE1), P-glycoprotein (P-gp) and multidrug resistance-associated protein 1 (MRP1). Flow cytometry detected cell cycle and apoptosis . Dual luciferase reporter gene assay and RNA immunoprecipitation (RIP) assay were used to confirm the targeting relationship between miR-155–5p, NORAD and WEE1. The xenograft model was used to observe the function of NORAD in vivo. immunohistochemistry (IHC) assay was used to detect the expression of WEE1, caspase-3 and Ki67 in tumor tissues. Results NORAD highly expressed in GEM-resistant BC cell lines . Knockdown of NORAD significantly inhibited the proliferation of T24/GEM cells, the expression of drug-resistant proteins P-gp and MRP1, inhibit the G0/G1 phase of cells, and induce cell apoptosis. Knockdown of NORAD reversed the promotion effect of miR-155–5p on WEE1 expression and promoted the sensitivity of T24/GEM cells to GEM. In vivo , knockdown of NORAD inhibited the tumor growth, and enhanced the GEM-sensitivity in mice. Conclusion These data highlight the potential of NORAD acts as a therapeutic target for BC GEM resistance. It revealed the vital roles of NORAD/miR-155–5p/WEE1 axis in GEM resistant BC cells, providing a novel therapeutic strategy for BC.
研究领域:
肿瘤学
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分子生物学
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药理学
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高影响因子
American Journal of Translational Research
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2021
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IF 4.06
Shan Jiang,Chonghe Wang,Jingyao Zhu,Xuezhu Huang
Objective: To investigate the regulatory mechanism of sevoflurane-induced neuronal apoptosis through analyzing the expression of glial cell-derived neurotrophic factor (GDNF) mediated by miR-133, sponged by long non-coding RNA (lncRNA) CDKN2B-AS1. Methods: An in vitro cell injury model was established by using different concentrations of sevoflurane and primary hippocampal neurons. Cell proliferation was detected by Cell Counting Kit-8 (CCK-8); caspase-3 and caspase-9 activities were detected by colorimetry, and apoptotic cells were determined by Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. Fluorescence in situ hybridization (FISH) analysis was used to detect localized expression of CDKN2B antisense RNA 1 (CDKN2B-AS1), and dual-luciferase reporter assay was employed to verify the correlation of CDKN2B-AS1 and miR-133, and of miR-133 and GDNF. The expression of CDKN2B-AS1, miR-133, and GDNF mRNA in the cell injury model were measured by quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Western blot was utilized to detect the expression of GDNF protein in the cell injury model. Results: In the cell injury model, CDKN2B-AS1 was highly expressed in the cytoplasm, and CDKN2B-AS1 and GDNF were downregulated and miR-133 was upregulated as detected by qRT-PCR (all P<0.05). The connections between CDKN2B-AS1 and miR-133, and between miR-133 and GDNF were confirmed. That is, CDKN2B-AS1 regulated the expression of GDNF by adsorbing miR-133 (all P<0.05). In cells treated with sevoflurane, overexpression of CDKN2B-AS1 could inhibit caspase-3 and caspase-9 activities and the degree of apoptosis. miR-133 could partially alleviate the effect of overexpressing CDKN2B-AS1 on cells, and si-GDNF the effect of miR-133 inhibitor (all P<0.05). Conclusion: lncRNA CDKN2B-AS1 can up-regulate the expression of GDNF, inhibit neuronal apoptosis, and ease the toxic effect of sevoflurane on neural cells by acting as a sponge to adsorb miR-133.
研究领域:
神经科学
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分子生物学
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细胞生物学
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高影响因子
Frontiers in Immunology
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2024
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IF 5.7
Jijin Jiang,Yan Chen,Yue Su,Li Zhang,Hao Qian,Xinmiao Song,Jin-Fu Xu
Background Sepsis is an uncontrolled reaction to infection that causes severe organ dysfunction and is a primary cause of ARDS. Patients suffering both sepsis and ARDS have a poor prognosis and high mortality. However, the mechanisms behind their simultaneous occurrence are unclear. Methods We acquired sepsis and ARDS datasets from GEO and Arrayexpress databases and screened hub genes by WGCNA and machine learning algorithm. For diagnosis and prognosis, ROC curve and survival analysis were used. We performed GO, KEGG, GSEA, immune cell infiltration, drug prediction, molecular docking, transcription factor prediction, and constructed PPI and ceRNA networks to explore these genes and the common mechanisms of sepsis and ARDS. Single-cell data analysis compared immune cell profiles and hub gene localization. Finally, RT-qPCR and H&E staining confirmed the reliability of hub genes using PBMCs samples and mouse models. Results We identified 242 common differentially expressed genes in sepsis and ARDS. WGCNA analysis showed that the turquoise module in GSE95233 is strongly linked to sepsis occurrence and poor prognosis, while the black module in GSE10474 is associated with ARDS. Using WGCNA and three machine learning methods (LASSO, random forest and Boruta), we identified three key genes CX3CR1, PID1 and PTGDS. Models built with them showed high AUC values in ROC curve evaluations and were validated by external datasets, accurately predicting the occurrence and mortality. We further explored the immunological landscape of these genes using immune infiltration and single-cell analysis. Then, the ceRNA, predicted drugs and molecular docking were analyzed. Ultimately, we demonstrated that these genes are expressed differently in human and mouse samples with sepsis and ARDS. Conclusion This study identified three molecular signatures (CX3CR1, PID1 and PTGDS) linked to the diagnosis and poor prognosis of sepsis and ARDS, validated by RT-qPCR and H&E staining in both patient and mouse samples. This research may be valuable for identifying shared biological mechanisms and potential treatment targets for both diseases.
研究领域:
医学遗传学
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免疫学
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生物信息学
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分子生物学
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转化医学
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npj Regenerative Medicine
·
2023
·Vol. 8
·期 1
·1-18
·
IF 7.2
Tu Tian,Shi Yuan,Zhou Boya,Wang Xiaoyu,Zhang Wenjie,Zhou Guangdong,Mo Xiumei,Wang Wenbo,Wu Jinglei,Liu Wei
Our previous work demonstrated the tendon-derived extracellular matrix (ECM) extracts as vital niches to specifically direct mesenchymal stem cells towards tenogenic differentiation. This study aims to further define the effective ECM molecules capable of teno-lineage induction on human adipose-derived stem cells (hASCs) and test their function for tendon engineering. By detecting the teno-markers expression levels in hASCs exposed to various substrate coatings, collagen I (COL1) and fibromodulin (FMOD) were identified to be the key molecules as a combination and further employed to the modification of poly(L-lactide- co -ε-caprolactone) electrospun nanoyarns, which showed advantages in inducting seeded hASCs for teno-lineage specific differentiation. Under dynamic mechanical loading, modified scaffold seeded with hASCs formed neo-tendon in vitro at the histological level and formed better tendon tissue in vivo with mature histology and enhanced mechanical properties. Primary mechanistic investigation with RNA sequencing demonstrated that the inductive mechanism of these two molecules for hASCs tenogenic differentiation was directly correlated with positive regulation of peptidase activity, regulation of cell-substrate adhesion and regulation of cytoskeletal organization. These biological processes were potentially affected by LOC101929398/has-miR-197-3p/TENM4 ceRNA regulation axis. In summary, COL1 and FMOD in combination are the major bioactive molecules in tendon ECM for likely directing tenogenic phenotype of hASCs and certainly valuable for hASCs-based tendon engineering.
研究领域:
组织工程
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干细胞生物学
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再生医学
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生物医学工程
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力学生物学
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