Journal: Cancer Cell International

Article Title: Hsa-miR-423-5p selectively loaded in hypoxic exosomes reduces the sensitivity of normoxic hepatocellular carcinoma to sorafenib via autophagy

doi: 10.1186/s12935-025-04055-8

Figure Lengend Snippet: HExs reduced the sensitivity of normoxic HCC cells to sorafenib via autophagy. A & B . Following pretreatment of the cells with exosomes, Annexin V-FITC/PI flow cytometry was used to determine the extent of sorafenib-induced apoptosis. C . The IC50 of sorafenib in Huh7 and 97H cells was detected using a CCK-8 assay. D . Huh7 and 97H cells were preincubated with or without HExs and subsequently treated with sorafenib (12, 24, and 48 h). The final assessment of cell viability was performed using a CCK-8 assay. E & F . After autophagy, sorafenib-induced apoptosis was detected via Annexin V-FITC/PI flow cytometry. G & H . Western blot analysis was used to detect the expression levels of autophagy markers in Huh7 and 97H cells that had been preincubated with or without hypoxic exosomes and subsequently treated with sorafenib (* p < 0.05, ** p < 0.01)

Article Snippet: The human hepatoma cell line Huh7, human hepatoma cell line MHCC-97H (97H) and human renal epithelial cell line 293 T were purchased from Procell Life Science & Technology Co., Ltd. (Wuhan, China).

Techniques: Flow Cytometry, CCK-8 Assay, Western Blot, Expressing

Journal: Cancer Cell International

Article Title: Hsa-miR-423-5p selectively loaded in hypoxic exosomes reduces the sensitivity of normoxic hepatocellular carcinoma to sorafenib via autophagy

doi: 10.1186/s12935-025-04055-8

Figure Lengend Snippet: Hsa-miR-423-5p was selectively loaded by HExs. A & B . Heatmaps and volcano plots of differentially expressed miRNAs between NExs and HExs secreted by Huh7 or 97H cells. C . Venn diagram of upregulated miRNAs in HExs secreted by Huh7 and 97H cells. D . The expression of the target miRNAs (hsa-miR-29a-3p and hsa-miR-1273f) in exosomes and cells was verified by RT‒qPCR. E & F . Heatmaps and volcano plots of differentially expressed miRNAs in Huh7 and 97H cells (hypoxic vs. normoxic) and exosomes (HExs vs. NExs). G . Venn diagram of miRNAs whose expression levels were not significantly changed in hypoxic HCC cells, downregulated in hypoxic HCC cells, or upregulated in HExs. H . The expression levels of hsa-miR-423-5p in exosomes and cells were verified by RT‒qPCR (N.S. p ≥ 0.05, * p < 0.05, ** p < 0.01, *** p < 0.001)

Article Snippet: The human hepatoma cell line Huh7, human hepatoma cell line MHCC-97H (97H) and human renal epithelial cell line 293 T were purchased from Procell Life Science & Technology Co., Ltd. (Wuhan, China).

Techniques: Expressing

Journal: Cancer Cell International

Article Title: Hsa-miR-423-5p selectively loaded in hypoxic exosomes reduces the sensitivity of normoxic hepatocellular carcinoma to sorafenib via autophagy

doi: 10.1186/s12935-025-04055-8

Figure Lengend Snippet: HNRNPA1 mediated the selective loading of hsa-miR-423-5p in HExs. A . The catRAPID website predicted RNA-binding proteins that could interact with hsa-miR-423-5p. B . HNRNPA1 expression was detected by Western blotting in Huh7 and 97H cells cultured under normoxic or hypoxic conditions. C . RNA pull-down experiments were performed using the miR-423-5p probe to isolate RNA-binding proteins, and the expression level of HNRNPA1 was subsequently assessed by Western blot analysis. D . RIP experiments were performed using an HNRNPA1-specific antibody to pull down miRNAs, and the expression level of hsa-miR-423-5p was subsequently measured by RT‒qPCR. E . Huh7 and 97H cells were infected with a specific knockdown lentivirus (sh-HNRNPA1) or a control lentivirus (sh-NC). The knockdown efficiency of sh-HNRNPA1 on HNRNPA1 was assessed by Western blotting. F . Cells were collected after infection with a lentivirus, along with their secreted exosomes. The expression levels of hsa-miR-423-5p in both cells and exosomes were measured by qRT-PCR (* p < 0.05, ** p < 0.01, *** p < 0.001)

Article Snippet: The human hepatoma cell line Huh7, human hepatoma cell line MHCC-97H (97H) and human renal epithelial cell line 293 T were purchased from Procell Life Science & Technology Co., Ltd. (Wuhan, China).

Techniques: RNA Binding Assay, Expressing, Western Blot, Cell Culture, Infection, Knockdown, Control, Quantitative RT-PCR

Journal: Cancer Cell International

Article Title: Hsa-miR-423-5p selectively loaded in hypoxic exosomes reduces the sensitivity of normoxic hepatocellular carcinoma to sorafenib via autophagy

doi: 10.1186/s12935-025-04055-8

Figure Lengend Snippet: Hsa-miR-423-5p promoted autophagy in normoxic HCC cells by targeting TAB2, thereby reducing the sensitivity of normoxic HCC cells to sorafenib . A . Huh7 and 97H cells were transfected with miR-423-5p mimics or mimics-NC, and the expression level of hsa-miR-423-5p in the cells was detected by RT‒qPCR. B . Expression levels of the autophagy markers LC3B and SQSTM1 in cells were detected by Western blotting. C . Huh7 and 97H cells were transfected with or without the miR-423-5p mimic, followed by sorafenib treatment, and apoptosis was detected via Annexin V-FITC/PI flow cytometry. D . Wild-type and mutated binding sites between hsa-miR-423-5p and TAB2. Relative luciferase intensity of cells cotransfected with miR-423-5p mimics or mimics-NC and TAB2-WT or TAB2-MUT. E . Expression level of TAB2 in Huh7 and 97H cells transfected with miR-423-5p mimics or mimics-NC (N.S. p ≥ 0.05, * p < 0.05, ** p < 0.01, *** p < 0.001)

Article Snippet: The human hepatoma cell line Huh7, human hepatoma cell line MHCC-97H (97H) and human renal epithelial cell line 293 T were purchased from Procell Life Science & Technology Co., Ltd. (Wuhan, China).

Techniques: Transfection, Expressing, Western Blot, Flow Cytometry, Binding Assay, Luciferase

Journal: Journal of Virology

Article Title: Ubiquitin-dependent proteasomal degradation of small hepatitis B virus surface antigen mediated by TRIM21 and antagonized by OTUD4

doi: 10.1128/jvi.02309-24

Figure Lengend Snippet: OTUD4 and TRIM21 alter the production of subviral particles and virions. (A, B, E, F) Lysates were prepared from Huh7 and HepG2 cells co-transfected with plasmids encoding 1.2HBV and either TRIM21-myc ( A, B ) or OTUD4-myc ( E, F ). These lysates were subjected to immunoblotting using antibodies against myc, β-actin, and SHBs, and intracellular HBV DNA levels were quantified by qPCR. Additionally, cell culture supernatants were analyzed by qPCR to quantify extracellular HBV DNA and virion DNA levels. (C, D, G, H) Lysates from cells transfected with plasmids encoding 1.2HBV and siRNA targeting either TRIM21 ( C, D ) or OTUD4 ( G, H ) were similarly processed. The lysates were immunoblotted using the indicated antibodies, and intracellular HBV DNA was quantified by qPCR. qPCR was performed on the supernatants to measure HBV DNA and virion DNA levels.

Article Snippet: The human hepatoma cell lines HepG2 and Huh7 were sourced from the American Type Culture Collection (Manassas, VA, USA) and the Japanese Collection of Research Bioresources Cell Bank (Osaka, Japan), respectively, while HUVECs were generously provided by Dr. Huang ( ).

Techniques: Transfection, Western Blot, Cell Culture

Journal: PLOS ONE

Article Title: Increased QPCT gene expression by the hepatitis B virus promotes HBV replication

doi: 10.1371/journal.pone.0312773

Figure Lengend Snippet: (B and C) HepG2.2.15 cells and HepG2 cells at the same cellular concentration (1 × 10 6 cells) were cultured for 48 h. Supernatants and cells were harvested. The protein and mRNA expression levels of QPCT were detected by Western blot and RT‒PCR, respectively. (E and F) After the gradient transfection of pBlue-HBV 1.3-fold plasmids into HepG2 cells, the cells were cultured for 48 h. Supernatants and cells were harvested. The protein and mRNA expression levels of QPCT were detected by Western blot and RT‒PCR, respectively, with the mRNA expression level of GAPDH as a reference. (H and I) Huh7 cells were analyzed in the same way as HepG2 cells. (A, D, and G) ELISA results for HBeAg and HBsAg concentrations in cell supernatants demonstrated successful infection.

Article Snippet: The human hepatoma cell line Huh7 was purchased from the China Center for Type Culture Collection (CCTCC).

Techniques: Concentration Assay, Cell Culture, Expressing, Western Blot, Transfection, Enzyme-linked Immunosorbent Assay, Infection

Journal: PLOS ONE

Article Title: Increased QPCT gene expression by the hepatitis B virus promotes HBV replication

doi: 10.1371/journal.pone.0312773

Figure Lengend Snippet: (A and D) HBV 1.3-fold plasmids and various concentrations of PcDNA3.1–3*Flag-QPCT were cotransfected into Huh7 and HepG2 cells and cultured for 48 h, supernatants and cells were harvested. (A and D) The overexpression of Flag-QPCT in the transfected HepG2 and Huh7 cells was detected by Western blot. (B and E) The expression levels of HBeAg and HBsAg in the supernatants were detected by ELISA. (C and F) Total RNA was extracted from the cells, and the mRNA expression of HBV pgRNA in the cells was detected by RT‒PCR, with the mRNA expression level of GAPDH as a reference. (G and H) Total DNA was extracted from the cells, and the HBV-DNA copies in Huh7 and HepG2 cells were detected by RT‒PCR.

Article Snippet: The human hepatoma cell line Huh7 was purchased from the China Center for Type Culture Collection (CCTCC).

Techniques: Cell Culture, Over Expression, Transfection, Western Blot, Expressing, Enzyme-linked Immunosorbent Assay