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: Chinese Medical Journal

Article Title: USP25 promotes hepatocellular carcinoma progression by interacting with TRIM21 via the Wnt/β-catenin signaling pathway

doi: 10.1097/CM9.0000000000002714

Figure Lengend Snippet: USP25 is highly expressed in HCC and is correlated with poor prognosis. (A) The expression of USP25 in pan-carcinoma as determined with the TIMER database. (B) The expression of USP25 mRNA was analyzed in HCC tissues from TCGA and ICGC databases. (C) The protein expression levels of USP25 in a normal liver cell line and six HCC cell lines. (D) The expression of USP25 in different HCC cell lines in the CCLE database. (E) Kaplan–Meier survival analysis of survival in HCC patients grouped according to USP25 expression ( n = 370). (F) The promoter methylation level of USP25 in different grades of HCC according to the analysis of patient samples. Grade 1: well differentiated; Grade 2: moderately-differentiated; Grade 3: moderately and poorly differentiated; Grade 4: poorly differentiated. (G) The expression of USP25 in HCC patients based on nodal metastasis status. * P <0.05; † P <0.001; ‡ P <0.01, § P <0.0001. ACC: Adrenocortieal carcinoma; BLCA: Bladder urothelial carcinoma; BRCA:Breast invasive carcinoma; CESC: Cervical squamous cell carcinoma; CHOL: Cholangiocarcinoma; COAD: Colon adenocarcinoma; DLBC: Lymphoid neoplasm diffuse large B-cell lymphoma; ESCA: Esophageal carcinoma; GBM: Glioblastoma multiforme; HNSC: head and neck squamous cell carcinoma; KICH: Kidney chromophobe; KIRC: Kidney renal clear cell carcinoma; KIRP: Kidney renal papillary cell carcinoma; LAML: Acute myeloid leukemia; LGG: Brain lower grade glioma; LIHC: Liver hepatocellular carcinoma; LUAD: Lung adenocarcinoma; LUSC: Lung squamous cell carcinoma; MESO: Mesothelioma; OV: Ovarian serous cystadenocarcinoma; PAAD: Pancreatic adenocarcinoma; PCPG: Pheochromocytoma and paraganglioma; PRAD: Prostate adenocarcinoma; READ: Rectum adenocarcinoma; SARC: Sarcoma; SKCM: Skin cutaneous melanoma; STAD: Stomach adenocarcinoma; TGCT: Testicular germ cell tumors; THCA: Thyroid carcinoma; THYM: Thymoma; UCEC: Uterine corpus endometrial carcinoma; UCS: Uterine carcinosarcoma; UVM: Uveal melanoma; HCC: Hepatocellular carcinoma; ICGC: International Cancer Genome Consortium; TIMER: Tumor Immune Estimation Resource; USP25: Ubiquitin-specific peptidase 25; TCGA: The Cancer Genome Atlas; CCLE: Cancer Cell Line Encyclopedia; HR: Hazard ratio; L-O2: Normal hepatocyte cell line; GAPDH: Glyceraldehyde-3-phosphate dehydrogenase; N0, 1: Node0,1.

Article Snippet: Six human HCC cell lines (HepG2, HuH7, SMMC7721, Bel7402, MHCC97H, and PLC/PRF/5) and a normal hepatocyte cell line (L-O2) were obtained from the China Center for Type Culture Collection (Wuhan, China).

Techniques: Expressing, Methylation, Ubiquitin Proteomics

Journal: Chinese Medical Journal

Article Title: USP25 promotes hepatocellular carcinoma progression by interacting with TRIM21 via the Wnt/β-catenin signaling pathway

doi: 10.1097/CM9.0000000000002714

Figure Lengend Snippet: USP25 promotes HCC cell proliferation in vitro. (A–C) HepG2 and MHCC97H cell lines with USP25 overexpression or knockdown were successfully established, as verified by analysis of protein and mRNA levels and fluorescence microscopy (Original magnification × 100). (D–F) Colony formation and CCK-8 assays were performed to detect cell proliferation. (G–J) DNA synthesis was measured by EdU IF staining (Original magnification × 200). * P <0.05; † P <0.001; ‡ P <0.01.CCK-8: Cell counting kit-8; HCC: Hepatocellular carcinoma; IF: Immunofluorescence; USP25: Ubiquitin-specific peptidase 25; EdU: 5-ethnyl-2-deoxyuridine; DAPI: 4′,6-Diamidino-2′-phenylindole; shUSP25: short hairpin Ubiquitin-specific peptidase 25; GAPDH: Glyceraldehyde-3-phosphate dehydrogenase.

Article Snippet: Six human HCC cell lines (HepG2, HuH7, SMMC7721, Bel7402, MHCC97H, and PLC/PRF/5) and a normal hepatocyte cell line (L-O2) were obtained from the China Center for Type Culture Collection (Wuhan, China).

Techniques: In Vitro, Over Expression, Knockdown, Fluorescence, Microscopy, CCK-8 Assay, DNA Synthesis, Staining, Cell Counting, Immunofluorescence, Ubiquitin Proteomics

Journal: Chinese Medical Journal

Article Title: USP25 promotes hepatocellular carcinoma progression by interacting with TRIM21 via the Wnt/β-catenin signaling pathway

doi: 10.1097/CM9.0000000000002714

Figure Lengend Snippet: USP25 promotes HCC cell migration and invasion in vitro. (A,B) A Transwell invasion assay was performed to assess the invasiveness of HepG2 and MHCC97H cells (Original magnification × 100). (C,D) A transwell migration assay was performed to analyze the migratory ability of HepG2 and MHCC97H cells (Original magnification × 100). (E–H) The scratch assay results demonstrated the migratory ability of HepG2 and MHCC97H cells (Original magnification × 40). (I,J) The expression levels of a PCNA and invasion-related proteins (MMP2 and MMP9) were detected by Western blotting. * P <0.001; † P <0.01. HCC: Hepatocellular carcinoma; PCNA: Proliferation-related protein; USP25: Ubiquitin-specific peptidase 25; MMP2: Matrix metallopeptidase 2; MMP9: Matrix metallopeptidase 9; shUSP25: short hairpin Ubiquitin-specific peptidase 25; GAPDH: Glyceraldehyde-3-phosphate dehydrogenase.

Article Snippet: Six human HCC cell lines (HepG2, HuH7, SMMC7721, Bel7402, MHCC97H, and PLC/PRF/5) and a normal hepatocyte cell line (L-O2) were obtained from the China Center for Type Culture Collection (Wuhan, China).

Techniques: Migration, In Vitro, Transwell Invasion Assay, Transwell Migration Assay, Wound Healing Assay, Expressing, Western Blot, Ubiquitin Proteomics

Journal: Chinese Medical Journal

Article Title: USP25 promotes hepatocellular carcinoma progression by interacting with TRIM21 via the Wnt/β-catenin signaling pathway

doi: 10.1097/CM9.0000000000002714

Figure Lengend Snippet: USP25 promotes HCC growth by activating the Wnt/β-catenin signaling pathway. (A) The expression of β-catenin was observed by IF staining (Original magnification × 200). (B) The mRNA expression of components in the Wnt/β-catenin signaling pathway was measured by qRT-PCR. (C) The expression of β-catenin was observed by Western blotting of nuclear-cytoplasmic fractionation. (D, E) Western blotting was used to detect the protein expression levels of Wnt/β-catenin pathway components and downstream factors. (F, G) Western blotting analysis after XAV939 treatment of HepG2 and MHCC97H cells. * P <0.001; † P <0.01; ‡ P <0.05, shUSP25 vs . Control; § P <0.05, shUSP25+XAV939 vs . Control. HCC: Hepatocellular carcinoma; IF: Immunofluorescence; qRT-PCR: Quantitative real-time polymerase chain reactio; USP25: Ubiquitin-specific peptidase 25.DAPI: 4′,6-Diamidino-2′-phenylindole; shUSP25: Short hairpin Ubiquitin-specific peptidase 25; PCNA: Proliferating cell nuclear antigen GAPDH: Glyceraldehyde-3-phosphate dehydrogenase; GSK3β: Glycogen synthase kinase 3 beta.

Article Snippet: Six human HCC cell lines (HepG2, HuH7, SMMC7721, Bel7402, MHCC97H, and PLC/PRF/5) and a normal hepatocyte cell line (L-O2) were obtained from the China Center for Type Culture Collection (Wuhan, China).

Techniques: Expressing, Staining, Quantitative RT-PCR, Western Blot, Fractionation, Control, Immunofluorescence, Ubiquitin Proteomics

Journal: BMB Reports

Article Title: Overexpression of CXCL2 inhibits cell proliferation and promotes apoptosis in hepatocellular carcinoma

doi: 10.5483/BMBRep.2018.51.12.140

Figure Lengend Snippet: Relative CXCL2 expression levels in hepatocellular carcinoma and normal tissues and its inhibition of cell proliferation in HCC cells. (A) In 264 pairs of tissues, CXCL2 expression was apparently down-regulated in tumours compared with adjacent liver tissues. ****P < 0.0001. (B) In the majority of HCC tissues (91%), CXCL2 mRNA levels were reduced. (C) Oncomine data ( www.oncomine.org ) showed under-expression of CXCL2 in HCC and paired normal liver tissues. ****P < 0.0001. (D) Immunohistochemical analysis of CXCL2 expression in three paired HCC and normal liver tissue. (E) Association between overall survival and CXCL2 expression in HCC patients. HCC, hepatocellular carcinoma. (F) CXCL2 expression was reduced in MHCC97H, HepG2, HCCLM3 and SMMC7721. (G) Western blot and RT-PCR confirmed CXCL2 mRNA and protein expression in HCCLM3-LV CXCL2, MHCC97H-LV CXCL2 and control group cells. (H) Representative images of EdU incorporation assays. **P < 0.01 and *P < 0.05. (I) Cell viability was analysed by the CCK-8 assay. ***P < 0.001.

Article Snippet: Six HCC cell lines (MHCC97H, SMMC7721, HCCLM3, Huh7, SKhep1, HepG2) and normal hepatocytes (QSG-7701) were purchased from the Liver Cancer Institute of Fudan University (Shanghai, China) and the American Type Culture Collection (Manassas, VA).

Techniques: Expressing, Inhibition, Immunohistochemical staining, Western Blot, Reverse Transcription Polymerase Chain Reaction, Control, CCK-8 Assay

Journal: BMB Reports

Article Title: Overexpression of CXCL2 inhibits cell proliferation and promotes apoptosis in hepatocellular carcinoma

doi: 10.5483/BMBRep.2018.51.12.140

Figure Lengend Snippet: The effects of CXCL2 overexpression on the cell cycle. (A) Representative FACS images of HCCLM3 and MHCC97H cells with LV-CXCL2 or LV-GFP. (B) Alterations in cell cycle distribution were analysed. **P < 0.01 and *P < 0.05. (C) Western blot analysis revealed the effect of CXCL2 on the expression of cyclin-dependent kinase and cyclin proteins. (D) Gene expression microarray showed 6908 differentially expressed genes. (E) The KEGG analysis indicated that CXCL2 regulated metabolic, PI3K—Akt signalling, cell cycle, pathways and so on. (F) CXCL2 reduced ERK1/2 phosphorylation in HCC cells.

Article Snippet: Six HCC cell lines (MHCC97H, SMMC7721, HCCLM3, Huh7, SKhep1, HepG2) and normal hepatocytes (QSG-7701) were purchased from the Liver Cancer Institute of Fudan University (Shanghai, China) and the American Type Culture Collection (Manassas, VA).

Techniques: Over Expression, Western Blot, Expressing, Gene Expression, Microarray, Phospho-proteomics

Journal: BMB Reports

Article Title: Overexpression of CXCL2 inhibits cell proliferation and promotes apoptosis in hepatocellular carcinoma

doi: 10.5483/BMBRep.2018.51.12.140

Figure Lengend Snippet: CXCL2 induces apoptosis in HCC cell lines and inhibits the growth of tumours in vivo . (A) Apoptosis rates of HCC cells transfected with CXCL2 compared with control as assessed by flow cytometry. (B) Quantitative analysis of apoptosis cells (Q2 + Q4, *P < 0.05). (C) Protein expression of pro-apoptosis mediators and anti-apoptosis regulators in HCC cells were determined by Western blot. (D) Photographic images of the subcutaneously formed tumour tissues. (E) LV-CXCL2-transfected HCCLM3 exhibited significantly reduced tumour volume and weight. *P < 0.05. (F) Representative images of CXCL2 protein expression as assessed by immunohistochemistry.

Article Snippet: Six HCC cell lines (MHCC97H, SMMC7721, HCCLM3, Huh7, SKhep1, HepG2) and normal hepatocytes (QSG-7701) were purchased from the Liver Cancer Institute of Fudan University (Shanghai, China) and the American Type Culture Collection (Manassas, VA).

Techniques: In Vivo, Transfection, Control, Flow Cytometry, Expressing, Western Blot, Immunohistochemistry

Journal: Cell Death & Disease

Article Title: Circular RNA circUBE2J2 acts as the sponge of microRNA-370-5P to suppress hepatocellular carcinoma progression

doi: 10.1038/s41419-021-04269-4

Figure Lengend Snippet: A The expression level of circUBE2J2 in the subcellular fractions of HepG2 cells was detected by qRT-PCR. NEAT1 and GAPDH were used as nuclear and cytoplasmic markers, respectively. B The overexpression and knockdown efficiency of circUBE2J2 were examined by qRT-PCR. C–E CCK-8 ( C ), colony formation assay ( D ), and EDU ( E ) were used to evaluate HCC cells proliferation after circUBE2J2 overexpression or knockdown. Scale bar: white bar, 200 μm. F Flow cytometry detection showing the percentages of cells in the G1, S, or G2 phase in both HepG2 and MHCC97H cells. G–L HCC cells were subcutaneously injected into nude mice, and tumor growth curves and tumor volume were plotted. Data are presented as mean ± SD; ** p < 0.05; ** p < 0.01; vs. NC. M Expression levels of Ki67 were observed in subcutaneous tumor tissues by IHC. The data are represented as the mean ± SD, n = 3. * p < 0.05; ** p < 0.01; NS no significance. OE overexpression; sh-circ transfected with lentivirus for circUBE2J2-specific shRNA; sh-NC transfected with control lentivirus.

Article Snippet: The human liver cancer cell lines MHCC97H were obtained from the Liver Cancer Institute of Fudan University; HepG2 and HLF were purchased from China Center for Type Culture Collection (CCTCC, Wuhan, China).

Techniques: Expressing, Quantitative RT-PCR, Over Expression, Knockdown, CCK-8 Assay, Colony Assay, Flow Cytometry, Injection, Transfection, shRNA, Control

Journal: Cell Death & Disease

Article Title: Circular RNA circUBE2J2 acts as the sponge of microRNA-370-5P to suppress hepatocellular carcinoma progression

doi: 10.1038/s41419-021-04269-4

Figure Lengend Snippet: A The wound healing of HCC cell. B Cell migration and invasion were assessed in both HepG2 and MHCC97H cells using transwell assays. C Western blot analysis for the relative levels of EMT-relevant protein expression in HepG2 and MHCC97H cells. D–I Tumor growth in HCC cells with the forced expression of circUBE2J2 was investigated by nude mice orthotopic tumor models. In orthotopic model mice, representative images of the liver ( D , F ). The size of the largest visible liver nodule was measured ( E , G ). Quantifications of visible surface liver metastatic nodules ( H , I ). Data are presented as mean ± SD; * p < 0.05; ** p < 0.01; NS no significance. OE overexpression; sh-circ transfected with lentivirus for circUBE2J2-specific shRNA; sh-NC transfected with control lentivirus.

Article Snippet: The human liver cancer cell lines MHCC97H were obtained from the Liver Cancer Institute of Fudan University; HepG2 and HLF were purchased from China Center for Type Culture Collection (CCTCC, Wuhan, China).

Techniques: Migration, Western Blot, Expressing, Over Expression, Transfection, shRNA, Control

Journal: Cell Death & Disease

Article Title: Circular RNA circUBE2J2 acts as the sponge of microRNA-370-5P to suppress hepatocellular carcinoma progression

doi: 10.1038/s41419-021-04269-4

Figure Lengend Snippet: A RAP assays were performed using an biotin probe against circUBE2J2 on extracts from HepG2 cells. Relative expression levels of circRNA were evaluated by qRT-PCR. B The relative expression level of potential target miRNA of circUBE2J2 was assessed by qRT-PCR. C , D RIP experiments were performed using an antibody against AGO2 on extracts from HLF and HepG2 cells. E Schematic diagram of reporter gene structure. F A schematic drawing showing the putative binding sites. The mutant version of circUBE2J2 is presented. G , H Relative luciferase activity determined 48 h after transfecting HEK293T cells ( G ) and HepG2 cells ( H ) with miR-370-5P mimic/NC or circUBE2J2 WT/Mut. WT wild type, Mut, mutant-type. I The expression of circUBE2J2 in MHCC97H, HLF, and HepG2 cells after transfection with miR-370-5P mimcs or miR-370-5P inhibitor. J The expression of miR-370-5P in HepG2, HLF, and MHCC97H cells after transfection with circUBE2J2-OE or circUBE2J2 shRNA. K RAP assay were performed using an probe against miR370-5P on extracts from HepG2 cells. Relative expression levels of circRNA were evaluated by qRT-PCR. L FISH assay analysis for the co-localization of miR-370-5P and circUBE2J2 in HCC cells. M miR-370-5P co-localized with circUBE2J2 in HCC adjacent non-tumor and tumor tissues was detected by FISH. Data are from three independent experiments (mean ± SEM). The data are represented as the mean ± SD, n = 3. * p < 0.05; ** p < 0.01; NS no significance.

Article Snippet: The human liver cancer cell lines MHCC97H were obtained from the Liver Cancer Institute of Fudan University; HepG2 and HLF were purchased from China Center for Type Culture Collection (CCTCC, Wuhan, China).

Techniques: Expressing, Quantitative RT-PCR, Binding Assay, Mutagenesis, Luciferase, Activity Assay, Transfection, shRNA

Journal: Cell Death & Disease

Article Title: Circular RNA circUBE2J2 acts as the sponge of microRNA-370-5P to suppress hepatocellular carcinoma progression

doi: 10.1038/s41419-021-04269-4

Figure Lengend Snippet: A , B The expression level of miR-370-5P in 40 TNBC tissues and their matched normal adjacent tissues was determined by qRT-PCR. C Overexpression and downregulation of miR-370-5P in hepatocellular carcinoma (HCC) cells using a miRNA-370-5P mimic or inhibitor, respectively, as measured by qRT-PCR. D miR-370-5P overexpression in MHCC97H and HLF cells promotes cell proliferation as measured by the CCK8 assay; downregulation of miR-370-5P in HepG2 cells inhibits cell proliferation as measured by the CCK8 assay. E The transwell assays. F The wound healing assays. G The expression levels of miR-370-5p were examined by qRT-PCR in HLF and MHCC97H cells with stable miR-370-5p overexpression. H , I HCC cells were subcutaneously injected into nude mice ( H ) and tumor volume were plotted ( I ). The data are represented as the mean ± SD, n = 3. * p < 0.05; ** p < 0.01; NS no significance. m miR370-5P mimcs, mn mimcs nc, i miR370-5P inhibitor, in inhibitor nc.

Article Snippet: The human liver cancer cell lines MHCC97H were obtained from the Liver Cancer Institute of Fudan University; HepG2 and HLF were purchased from China Center for Type Culture Collection (CCTCC, Wuhan, China).

Techniques: Expressing, Quantitative RT-PCR, Over Expression, CCK-8 Assay, Injection

Journal: Cell Death & Disease

Article Title: Circular RNA circUBE2J2 acts as the sponge of microRNA-370-5P to suppress hepatocellular carcinoma progression

doi: 10.1038/s41419-021-04269-4

Figure Lengend Snippet: A Venn diagram analyses of three independent databases reveals 72 possible targets of miR370-5P. B Pearson’s correlation analysis showed that miR-370-5P was inversely correlated with KLF7 mRNA levels. C Overexpression and downregulation of miR-370-5P in MHCC97H,HLF and HepG2 cells changes the KLF7 mRNA level as measured by qRT-PCR. D Bioinformatics predicted binding sites of miR-370-5P within KLF7 were shown. E A schematic drawing showing the putative binding sites. The mutant version of KLF7 is presented. F , G Relative luciferase activity determined 48 h after transfecting HEK293T cells ( D ) and HLF cells ( E ) with miR-370-5P mimic/NC or KLF7 WT/Mut. Data are presented as means ± SD, n = 3. ** p < 0.05; ** p < 0.01. WT wild-type, Mut mutant-type, m miR370-5P mimcs, mn mimcs nc, i miR370-5P inhibitor, in inhibitor nc.

Article Snippet: The human liver cancer cell lines MHCC97H were obtained from the Liver Cancer Institute of Fudan University; HepG2 and HLF were purchased from China Center for Type Culture Collection (CCTCC, Wuhan, China).

Techniques: Over Expression, Quantitative RT-PCR, Binding Assay, Mutagenesis, Luciferase, Activity Assay

Journal: Cell Death & Disease

Article Title: Circular RNA circUBE2J2 acts as the sponge of microRNA-370-5P to suppress hepatocellular carcinoma progression

doi: 10.1038/s41419-021-04269-4

Figure Lengend Snippet: A qRT-PCR detection show the differential expression of KLF7 in 40 paired HCC tissues. B qRT-PCR detection showing the expression of KLF7 in both MHCC97H and HepG2 cells after transfection with siRNA against KLF7 or negative control. C CCK8 assays were used to evaluate cell proliferation. D Colony formation assay showing proliferation in both MHCC97H and HepG2 cells. E The migration of HCC cells. F The wound-healing assays. G Flow cytometry detection showing the percentages of cells in G1, S, or G2 phase in both MHCC97H and HepG2 cells. Data are presented as mean ± SD, n = 3. * p < 0.05, ** p < 0.01; NS no significance.

Article Snippet: The human liver cancer cell lines MHCC97H were obtained from the Liver Cancer Institute of Fudan University; HepG2 and HLF were purchased from China Center for Type Culture Collection (CCTCC, Wuhan, China).

Techniques: Quantitative RT-PCR, Quantitative Proteomics, Expressing, Transfection, Negative Control, Colony Assay, Migration, Flow Cytometry

Journal: Cell Death & Disease

Article Title: Circular RNA circUBE2J2 acts as the sponge of microRNA-370-5P to suppress hepatocellular carcinoma progression

doi: 10.1038/s41419-021-04269-4

Figure Lengend Snippet: A KLF7 expression in the mRNA level was measured by qPCR in HCC cells after circUBE2J2 knockdown. B After circUBE2J2 knocking down, miR-370-5P inhibitor or inhibitor nc were added to HLF and HepG2 cell culture. The KLF7 protein expression level was analyzed by western blot. C The cell proliferation in HCC cells with the reduced expression of miR-370-5P was assessed by a CCK-8 assay. D The proliferation ratio of HCC cells was measured by EdU incorporation. E The migration abilities in HCC cells with the reduced expression of miR-370-5P were evaluated via a transwell assay. F After miR-370-5P overexpression, KLF7 overexpression vector was added to MHCC97H and HLF cell culture. KLF7 expression in the mRNA level was measured by qPCR. G The KLF7 protein expression level was analyzed by western blot. H The cell proliferation in HCC cells was assessed by a CCK-8 assay. I Colony formation assays show the proliferation. J Transwell migration assays. Data are presented as mean ± SD, n = 3. * p < 0.05, ** p < 0.01; NS no significance. sh-circ transfected with lentivirus for circUBE2J2-specific shRNA; sh-NC transfected with control lentivirus.

Article Snippet: The human liver cancer cell lines MHCC97H were obtained from the Liver Cancer Institute of Fudan University; HepG2 and HLF were purchased from China Center for Type Culture Collection (CCTCC, Wuhan, China).

Techniques: Expressing, Knockdown, Cell Culture, Western Blot, CCK-8 Assay, Migration, Transwell Assay, Over Expression, Plasmid Preparation, Transfection, shRNA, Control