LABORATORY INVESTIGATION
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2023
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IF 5
Wei Zhao,Jing Zhao,Kun Li,Yanjiao Hu,Dongxia Yang,Bin Tan,Jian Shi
Nuclear factor of activated T cells 2 (NFATC2) is reported to contribute to the initiation and progression of various cancers; however, its expression and function in cholangiocarcinoma (CCA) remain elusive. Herein, we investigated the expression pattern, clinicopathological characteristics, cell biological functions, and potential mechanisms of NFATC2 in CCA. Real-time PCR and immunohistochemistry were performed to analyze the expression of NFATC2 in human CCA tissues. CCK-8, colony formation, flow cytometry, Western blotting and transwell assays, and in vivo xenograft and pulmonary metastasis models were used to explore the effect of NFATC2 on the proliferation and metastasis of CCA. A dual-luciferase reporter system, oligonucleotide pull down, chromatin immunoprecipitation, immunofluorescence, and coimmunoprecipitation were performed to reveal the potential mechanisms. We found that NFATC2 was upregulated in CCA tissues and cells, and its aberrantly high levels were associated with poorer differentiation pattern. Functionally, NFATC2 overexpression promoted CCA cell proliferation and metastasis, whereas knockdown of NFATC2 led to the opposite results. Mechanistically, NFATC2 could be enriched in the promoter region of neural precursor cell expressed developmentally downregulated 4 (NEDD4) to facilitate its expression. Furthermore, NEDD4 targeted fructose-1, 6-bisphosphatase 1 (FBP1), and inhibited FBP1 expression via ubiquitination. In addition, silencing NEDD4 rescued the effects of NFATC2 overexpression on CCA cells. NEDD4 was upregulated in human CCA tissues, and its expression levels were positively correlated with those of NFATC2. We thus conclude that NFATC2 promotes the progression of CCA via the NEDD4/FBP1 axis, emphasizing the oncogenic role of NFATC2 in CCA progression.
研究领域:
肿瘤学
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分子生物学
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细胞生物学
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癌症研究
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DOI
PubMed
高影响因子
BMC CANCER
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2021
·Vol. 21
·期 1
·1-14
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IF 4.43
Zhao Wei,Zhao Jing,Guo Xiao,Feng Yujie,Zhang Bingyuan,Tian Lantian
Background Cholangiocarcinoma is a common malignant tumor of digestive system. LncRNA metallothionein 1 J, pseudogene (MT1JP) has been reported to play tumor-suppressing roles in multiple cancers. However, its effect on cholangiocarcinoma has not been evaluated. Methods The expression of MT1JP in intrahepatic cholangiocarcinoma specimens and paired para-carcinoma tissues were detected by real-time PCR. The overexpression plasmid and siRNA of MT1JP were transfected into intrahepatic cholangiocarcinoma cells to change the expression levels of MT1JP. CCK-8, flow cytometry and transwell assays were performed to measure proliferation, cell cycle transition, apoptosis, migration and invasion. Dual-luciferase reporter assay, real-time PCR and western blot were carried out to screen the miRNA bound by MT1JP. In addition, xenograft experiment was used to determine the tumorigenesis of cholangiocarcinoma cells in nude mice. Results MT1JP was downregulated in intrahepatic cholangiocarcinoma specimens, and its expression was related with TNM stage and lymph node metastasis. Overexpression of MT1JP inhibited proliferation, cell cycle transition, migration and invasion, and induced apoptosis in intrahepatic cholangiocarcinoma cells. The knockdown of MT1JP led to opposite results. MT1JP bound to miR-18a-5p to facilitate the expression of fructose-1,6-bisphosphatase 1 (FBP1). MiR-18a-5p was increased in intrahepatic cholangiocarcinoma samples, and its expression was negatively correlated with that of MT1JP. In addition, MT1JP also suppressed tumorigenesis in nude mice. Conclusions MT1JP alleviated proliferation, migration and invasion, and induced apoptosis in cholangiocarcinoma cells by regulating miR-18a-5p/FBP1 axis. These findings may provide novel insights for clinical diagnosis and treatment of cholangiocarcinoma.
研究领域:
肿瘤学
·
分子生物学
·
癌症研究
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产品关联
DOI
PubMed
高影响因子
PHYTOMEDICINE
·
2025
·
IF 8.3
Kaili Liu,Jianli Li,Zhiwei Sun,Yuheng Sun,Xuerui Zhang,Yang Sui,Zhongyuan Qu,Xiang Zou
Background Chelidonium majus L., a traditional herbal medicine listed in the Pharmacopoeia of China (2025), has remarkable anticancer properties. Chelidonine (CHE), its primary bioactive alkaloid, can inhibit the proliferation of breast cancer (BC) cells; however, whether its antitumor effect in BC is mediated through regulating M2 macrophage polarization remains unexplored. Purpose To investigate the effect of CHE on M2 macrophage polarization in BC and to identify key molecular targets and pathways involved in its immunomodulatory effects. Methods The effect of CHE on tumour progression was evaluated in vivo using a 4T1 BC mouse model. In addition, the effects of CHE on M2 macrophage polarization were investigated both in vitro and in vivo via immunofluorescence and Western blotting. Coculture assays were conducted to elucidate the mechanism by which CHE exerts antiproliferative and antimetastatic effects through the inhibition of M2 macrophage polarization. An integrated analysis of network pharmacology, single-cell, and bulk RNA-seq data was performed to identify potential molecular targets of CHE in the suppression of M2 macrophage polarization. Molecular docking, Western blotting, and ELISA were utilized for target validation to elucidate the roles in M2 macrophage polarization. Results CHE effectively impeded tumour progression in BC mice. Both in vitro and in vivo experiments demonstrated that CHE significantly inhibited M2 macrophage polarization, as evidenced by the altered expression of key polarization markers, including CD86, CD206, Arg-1, and CD11c. Under coculture conditions, CHE disrupted the mitochondrial membrane potential and compromised membrane integrity in BC cells, triggering pyroptosis via activation of the NLRP3/Caspase-1/GSDMD signaling axis. Additionally, CHE attenuated the epithelial–mesenchymal transition (EMT) process, thus reducing the migratory and invasive capacities of BC cells. An integrated analysis of network pharmacology, single-cell, and bulk RNA sequencing data revealed FBP1 as a potential molecular target through which CHE suppresses M2 macrophage polarization. Molecular docking studies revealed a relatively strong binding affinity between CHE and FBP1. Experimental validation further confirmed that CHE downregulated the expression of FBP1 and its downstream target p-STAT3 in macrophages, thereby inhibiting M2 polarization. Conclusions CHE might suppress M2 polarization and tumour progression by targeting FBP1. These findings highlight its potential as a therapeutic agent in immunotherapy and BC treatment.
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产品关联
DOI
PubMed
高影响因子
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