Journal: Cell Communication and Signaling : CCS

Article Title: Integrated analysis reveals critical glycolytic regulators in hepatocellular carcinoma

doi: 10.1186/s12964-020-00539-4

Figure Lengend Snippet: OPN promotes the Warburg effect in HCC cells. a The knockdown efficiency of OPN in HCC-LM3 cells was measured by Western blotting and ELISA. b Effects of OPN knockdown on the glucose uptake and lactate production in HCC-LM3 cells ( n = 3). c The extracellular acidification rate (ECAR) in sh-OPN and sh-Ctrl HCC-LM3 cells was measured by Seahorse analyzer ( n = 5). d Effects of OPN blockade on the glucose uptake and lactate production in HCC-LM3 cells ( n = 3). e The overexpression efficiency of OPN in NIH3T3 cells and MEFs was measured by Western blotting. f Effects of OPN overexpression on the glucose uptake and lactate production in NIH3T3 cells and MEFs ( n = 3). g Effects of OPN overexpression on ECAR in NIH3T3 cells and MEFs were measured by Seahorse analyzer ( n = 5). * P < 0.05 and ** P < 0.01

Article Snippet: The human HCC cell lines (Huh7, HCC-LM3, SMCC-7721, MHCC-97H, Hep3B, and MHCC-97H), NIH3T3 cells, and MEFs were obtained from American Type Culture Collection (ATCC, UK).

Techniques: Knockdown, Western Blot, Enzyme-linked Immunosorbent Assay, Over Expression

Journal: Cell Communication and Signaling : CCS

Article Title: Integrated analysis reveals critical glycolytic regulators in hepatocellular carcinoma

doi: 10.1186/s12964-020-00539-4

Figure Lengend Snippet: Certification of the negative regulators of HCC glycolysis. a Western blotting showed the overexpression efficiency of SPP2, LECT2, SLC10A1, CYP3A4, HSD17B13, and IYD in Huh7 cells. b Real-time qPCR analysis showed the overexpression efficiency of SPP2, LECT2, SLC10A1, CYP3A4, HSD17B13, and IYD in Huh7 cells ( n = 3). c-e Measurement of SPP2, LECT2, SLC10A1, CYP3A4, HSD17B13, or IYD overexpression on the glucose utilization ( f , n = 3), lactate production ( g , n = 3) and ECAR ( h , n = 5) in Huh7 cells. * P < 0.05, ** P < 0.01, and *** P < 0.001

Article Snippet: The human HCC cell lines (Huh7, HCC-LM3, SMCC-7721, MHCC-97H, Hep3B, and MHCC-97H), NIH3T3 cells, and MEFs were obtained from American Type Culture Collection (ATCC, UK).

Techniques: Western Blot, Over Expression

Journal: Cell Communication and Signaling : CCS

Article Title: Integrated analysis reveals critical glycolytic regulators in hepatocellular carcinoma

doi: 10.1186/s12964-020-00539-4

Figure Lengend Snippet: Effects of glycolysis-related genes on HCC tumor growth. a Colony formation assay showed that OPN knockdown or blockade inhibits HCC-LM3 cell proliferation ( n = 3). b Colony formation assay for Huh3B cells treated with recombinant OPN protein ( n = 3). c Colony formation assay showed that SPP2, LECT2, SLC10A1, CYP3A4, HSD17B13, or IYD overexpression inhibits Huh7 cell proliferation ( n = 3). d The effects of glycolysis-related genes on HCC tumor growth in the presence or absence of 5 mM 2-DG ( n = 3). e In the culture medium containing 25 mM glucose or galactose, the effects of glycolysis-related genes on HCC tumor growth were analyzed by clonogenic assay. * P < 0.05 and ** P < 0.01

Article Snippet: The human HCC cell lines (Huh7, HCC-LM3, SMCC-7721, MHCC-97H, Hep3B, and MHCC-97H), NIH3T3 cells, and MEFs were obtained from American Type Culture Collection (ATCC, UK).

Techniques: Colony Assay, Knockdown, Recombinant, Over Expression, Clonogenic Assay

Journal: Cell Communication and Signaling : CCS

Article Title: Integrated analysis reveals critical glycolytic regulators in hepatocellular carcinoma

doi: 10.1186/s12964-020-00539-4

Figure Lengend Snippet: OPN promotes HCC glycolysis by modulating αvβ3-NF-κB signaling. a Blockade of integrin αvβ3 with Cilengitide inhibits glucose utilization ( n = 3), lactate production ( n = 3) and ECAR ( n = 5) in HCC-LM3 cells. b Western blotting analysis the signaling pathway influenced by OPN. c Glucose utilization and lactate production in Hep3B cells upon treatment with OPN recombinant protein and indicated pathway inhibitors ( n = 3). d Effect of OPN on the NF-κB activity in HCC cells ( n = 3). e Effect of CA-IKKβ on the glucose uptake and lactate production in OPN-silenced HCC-LM3 cells ( n = 3). * P < 0.05 and ** P < 0.01; ns: not significant

Article Snippet: The human HCC cell lines (Huh7, HCC-LM3, SMCC-7721, MHCC-97H, Hep3B, and MHCC-97H), NIH3T3 cells, and MEFs were obtained from American Type Culture Collection (ATCC, UK).

Techniques: Western Blot, Recombinant, Activity Assay

Journal: Cell Communication and Signaling : CCS

Article Title: Integrated analysis reveals critical glycolytic regulators in hepatocellular carcinoma

doi: 10.1186/s12964-020-00539-4

Figure Lengend Snippet: Inhibition of OPN-αvβ3 axis suppresses HCC tumor growth and glycolyis. a Tumor volume in sh-Ctrl and sh-OPN HCC-LM3 xenografts as indicated time point was measured ( n = 5). b Effect of Cilengitide treatment on the tumor growth of HCC-LM3 xenografts ( n = 5). c The lactate level in the tumor tissues from ( a ) and ( b ) was detected ( n = 5). d The expression of glycolytic genes in the tumor tissues from ( a ) and ( b ) was analyzed by real-time qPCR ( n = 5). e Hematoxylin and eosin staining in liver tissue samples from tumor-bearing WT and OPN-KO mice. f The expression of glycolytic genes in liver tissue samples from tumor-bearing WT and OPN-KO mice was analyzed by real-time qPCR ( n = 5). * P < 0.05 and ** P < 0.01

Article Snippet: The human HCC cell lines (Huh7, HCC-LM3, SMCC-7721, MHCC-97H, Hep3B, and MHCC-97H), NIH3T3 cells, and MEFs were obtained from American Type Culture Collection (ATCC, UK).

Techniques: Inhibition, Expressing, Staining

Journal: Cell Communication and Signaling : CCS

Article Title: Integrated analysis reveals critical glycolytic regulators in hepatocellular carcinoma

doi: 10.1186/s12964-020-00539-4

Figure Lengend Snippet: Expression pattern of OPN in clinical samples. a The expression of glycolytic genes in human HCC tissue samples with high OPN ( n = 10) and low OPN ( n = 20) expression was analyzed by real-time qPCR. b Representative photographs of OPN expression in HCC tumor tissues; scale bar: 50 μm. The correlation between OPN expression and the SUVmax value was analyzed. * P < 0.05 and ** P < 0.01

Article Snippet: The human HCC cell lines (Huh7, HCC-LM3, SMCC-7721, MHCC-97H, Hep3B, and MHCC-97H), NIH3T3 cells, and MEFs were obtained from American Type Culture Collection (ATCC, UK).

Techniques: Expressing

Journal: Journal of Immunology Research

Article Title: Exosomal miR-452-5p Induce M2 Macrophage Polarization to Accelerate Hepatocellular Carcinoma Progression by Targeting TIMP3

doi: 10.1155/2022/1032106

Figure Lengend Snippet: miR-452-5p inhibition suppressed HCC cell migration and invasion. (a) miR-452-5p expression in HCC cells and normal human epithelial cells. (b) miR-452-5p was successfully inhibited by the miR-inhibitor. (c, d) CCK-8 assay and EdU staining of HCC cells with and without miR-452-5p inhibition. Optical density (OD) was measured at 24, 48, and 72 h after transfection. (e) Apoptosis rate was analyzed by flow cytometry. (f, g) Migration and invasion were detected by Transwell assay. ∗∗ P < 0.01.

Article Snippet: HCC cells (Huh-7, MHCC97-L, HCCLM3, and SNU-182), normal human epithelial cell THLE-3, and THP-1 monocytes were obtained from ATCC.

Techniques: Inhibition, Migration, Expressing, CCK-8 Assay, Staining, Transfection, Flow Cytometry, Transwell Assay

Journal: Journal of Immunology Research

Article Title: Exosomal miR-452-5p Induce M2 Macrophage Polarization to Accelerate Hepatocellular Carcinoma Progression by Targeting TIMP3

doi: 10.1155/2022/1032106

Figure Lengend Snippet: miR-452-5p mainly reside in HCC cells-derived exosomes. (a) miR-452-5p in the culture medium of normal epithelial cells and HCC cells. (b) miR-452-5p were encapsulated protected from RNase. (c, d) TEM and WB validation of purified exosomes from SNU-182 and Huh-7 cells. (e) miR-452-5p in HCC cells treated with GW4869 or purified exosomes are analyzed by qRT-PCR. ∗∗ P < 0.01. Scale bar: 200 nm.

Article Snippet: HCC cells (Huh-7, MHCC97-L, HCCLM3, and SNU-182), normal human epithelial cell THLE-3, and THP-1 monocytes were obtained from ATCC.

Techniques: Derivative Assay, Biomarker Discovery, Purification, Quantitative RT-PCR

Journal: Journal of Immunology Research

Article Title: Exosomal miR-452-5p Induce M2 Macrophage Polarization to Accelerate Hepatocellular Carcinoma Progression by Targeting TIMP3

doi: 10.1155/2022/1032106

Figure Lengend Snippet: HCC cells deserved exosomal miR-452-5p induces M2 polarization of macrophages. (a) PKH26-labelled SNU-182-exo and Huh-7-exo. (b) mRNA expression of macrophage markers after coculturing with HCC cell exosome. (c) mRNA expression of M2 macrophage markers. (d) mRNA expression of M2 macrophage markers in macrophages treated with exosomes from miR-452-5p inhibited or overexpressed HCC cells. ∗∗ P < 0.01. Scale bar: 100 μ m.

Article Snippet: HCC cells (Huh-7, MHCC97-L, HCCLM3, and SNU-182), normal human epithelial cell THLE-3, and THP-1 monocytes were obtained from ATCC.

Techniques: Expressing

Journal: Journal of Immunology Research

Article Title: Exosomal miR-452-5p Induce M2 Macrophage Polarization to Accelerate Hepatocellular Carcinoma Progression by Targeting TIMP3

doi: 10.1155/2022/1032106

Figure Lengend Snippet: HCC cells deserved exosomal miR-452-5p accelerates M2 macrophage polarization to stimulate HCC cell migration, invasion in vitro , and tumorigenesis in vivo . (a) Migration and invasion rates of transfected MHCC97-L cells, M-PBS set up as a negative control. (b) Tumorigenicity of xenograft mice models. (c) Tumor volumes were measured each week for three weeks. (d) Tumor weights were measured after mice were sacrificed. ∗∗ P < 0.01.

Article Snippet: HCC cells (Huh-7, MHCC97-L, HCCLM3, and SNU-182), normal human epithelial cell THLE-3, and THP-1 monocytes were obtained from ATCC.

Techniques: Migration, In Vitro, In Vivo, Transfection, Negative Control

Journal: Mediators of Inflammation

Article Title: miR-559 Inhibits Proliferation, Autophagy, and Angiogenesis of Hepatocellular Carcinoma Cells by Targeting PARD3

doi: 10.1155/2022/3121492

Figure Lengend Snippet: miR-559 was downregulated, and PARD3 was upregulated in HCC cells. (a) The expression level of miR-559 in HCC cells. (b) The mRNA expression of PARD3 in HCC cells. (c) The protein level of PARD3 in HCC cells. The values were obtained from three independent experiments and shown as mean ± SD. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001, compared with L02 cells.

Article Snippet: The HCC cell lines (SNU-387, Huh-7, HCCLM3, and MHCC-97H cells) and the normal human liver cell line L02 were provided by ATCC (Manassas, VA, USA).

Techniques: Expressing

Journal: Frontiers in Pharmacology

Article Title: Iberverin exhibits antineoplastic activities against human hepatocellular carcinoma via DNA damage-mediated cell cycle arrest and mitochondrial-related apoptosis

doi: 10.3389/fphar.2023.1326346

Figure Lengend Snippet: Iberverin inhibits the viability and proliferation of HCC cells. (A) The chemical structure formula of iberverin. (B) HCCLM3, HepG2, Huh1, Huh7, Huh7.5.1, SMMC7721 and SNU739 cells were incubated with iberverin for 48 h. The 50% inhibiting concentrations (IC 50 ) were calculated based on the MTT assay ( n = 3). (C) Colony formation assay of Huh7, Huh7.5.1 and SNU739 cells incubated with 10, 20, and 40 μmol/L iberverin or DMSO control for 2 weeks ( n = 3). Data were shown as mean ± SD (standard deviation). ** p < 0.01, *** p < 0.001 and **** p < 0.0001.

Article Snippet: The human HCC cell lines HCCLM3, HepG2, Huh7, Huh7.5.1 and SMMC7721 were purchased from the China Typical Culture Collection Center (CCTCC), SNU739 from the Korean Cell Line Bank (KCLB), and Huh1 from the Japanese Collection of Research Bioresources Cell Bank (JCRB).

Techniques: Incubation, MTT Assay, Colony Assay, Control, Standard Deviation