Journal: Cell Transplantation

Article Title: circRNA hsa_circ_104566 Sponged miR-338-3p to Promote Hepatocellular Carcinoma Progression

doi: 10.1177/0963689720963948

Figure Lengend Snippet: Expression of hsa_circ_104566 was upregulated in HCC. (A) The expression levels of circRNA hsa_circ_104566 in HCC tissues, including high pathological grade (TNM III and IV) and low grade (TNM I and II), and adjacent noncancer tissues measured by qRT-PCR ( N = 87). (B) Overall survival analysis of HCC patients with high and low levels of hsa_circ_104566 expression. (C) The expression levels of hsa_circ_104566 in HCC cell lines (SK-HEP-1, HLE, SNU449, Hep-3B, and Huh7) and MIHA measured by qRT-PCR. (D) Hsa_circ_104566 was resistant to RNase R digestion compared with linear PSD3 in both Huh7 and SK-HEP-1 cells. (E) Expression levels of hsa_circ_104566, GAPDH, and U6 in cytoplasm or nucleus of Huh7 and SK-HEP-1 cells measured by qRT-PCR, suggesting the mainly cytoplasm subcellular localization of hsa_circ_104566. circRNA: circular RNA; GAPDH: glyceraldehyde 3-phosphate dehydrogenase; HCC: hepatocellular carcinoma; qRT-PCR: quantitative real-time polymerase chain reaction; TNM: tumor node metastasis.

Article Snippet: HCC cell lines (SK-HEP-1, HLE, SNU449, Hep-3B, Huh7) and MIHA normal liver cell line, obtained from the Chinese Academy of Sciences (Shanghai, China), were cultured in Roswell Park Memorial Institute Medium (RPMI) 1640 medium (Lonza, Basel, Switzerland) supplemented with 10% fetal bovine serum (FBS; Gibco, Waltham, MA, USA) at 37 °C constant temperature and incubated with 5% carbon dioxide.

Techniques: Expressing, Quantitative RT-PCR, Real-time Polymerase Chain Reaction

Journal: Cell Transplantation

Article Title: circRNA hsa_circ_104566 Sponged miR-338-3p to Promote Hepatocellular Carcinoma Progression

doi: 10.1177/0963689720963948

Figure Lengend Snippet: Hsa_circ_104566 promoted cell migration and invasion of hepatocellular carcinoma (HCC). (A) The influence of hsa_circ_104566 on cell migration of Huh7 and SK-HEP-1 cells measured by wound healing assay. (B) The influence of hsa_circ_104566 on cell invasion of Huh7 and SK-HEP-1 cells measured by Transwell assay. (C) The influence of hsa_circ_104566 on protein expression of E-cadherin and N-cadherin in Huh7 and SK-HEP-1 cells measured by immunofluorescence.

Article Snippet: HCC cell lines (SK-HEP-1, HLE, SNU449, Hep-3B, Huh7) and MIHA normal liver cell line, obtained from the Chinese Academy of Sciences (Shanghai, China), were cultured in Roswell Park Memorial Institute Medium (RPMI) 1640 medium (Lonza, Basel, Switzerland) supplemented with 10% fetal bovine serum (FBS; Gibco, Waltham, MA, USA) at 37 °C constant temperature and incubated with 5% carbon dioxide.

Techniques: Migration, Wound Healing Assay, Transwell Assay, Expressing, Immunofluorescence

Journal: Cell Transplantation

Article Title: circRNA hsa_circ_104566 Sponged miR-338-3p to Promote Hepatocellular Carcinoma Progression

doi: 10.1177/0963689720963948

Figure Lengend Snippet: Hsa_circ_104566 bound miR-338-3p in HCC cells. (A) miR-338-3p was predicted as the potential binding target of hsa_circ_104566 via circular RNA Interactome ( https://circinteractome.nia.nih.gov/ ). (B) The influence of miR-338-3p mimics on luciferase activities of pmirGLO-wt-hsa_circ_104566 or pmirGLO-mut-hsa_circ_104566 in Huh7 and SK-HEP-1 cells. (C) Subcellular localization of hsa_circ_104566 and miR-338-3p in Huh7 and SK-HEP-1 cells examined via RNA-fluorescence in situ hybridization. (D) The influence of hsa_circ_104566 on miR-338-3p expression in Huh7 and SK-HEP-1 cells. (E) The expression of miR-338-3p in HCC tissues and adjacent noncancer tissues measured by qRT-PCR ( N = 87). (F) Negative correlation between miR-338-3p and hsa_circ_104566 in HCC patients. HCC: hepatocellular carcinoma; qRT-PCR: quantitative real-time polymerase chain reaction.

Article Snippet: HCC cell lines (SK-HEP-1, HLE, SNU449, Hep-3B, Huh7) and MIHA normal liver cell line, obtained from the Chinese Academy of Sciences (Shanghai, China), were cultured in Roswell Park Memorial Institute Medium (RPMI) 1640 medium (Lonza, Basel, Switzerland) supplemented with 10% fetal bovine serum (FBS; Gibco, Waltham, MA, USA) at 37 °C constant temperature and incubated with 5% carbon dioxide.

Techniques: Binding Assay, Luciferase, Fluorescence, In Situ Hybridization, Expressing, Quantitative RT-PCR, Real-time Polymerase Chain Reaction

Journal: Cell Transplantation

Article Title: circRNA hsa_circ_104566 Sponged miR-338-3p to Promote Hepatocellular Carcinoma Progression

doi: 10.1177/0963689720963948

Figure Lengend Snippet: miR-338-3p bound FOXP1 in HCC cells. (A) FOXP1 was predicted as a potential miR-338-3p binding targets via Targetscan ( http://www.targetscan.org/vert_71/ ). (B) The influence of miR-338-3p mimics on luciferase activities of pmirGLO-wt-FOXP1 or pmirGLO-mut-FOXP1 in Huh7 and SK-HEP-1 cells. (C) The influence of hsa_circ_104566 on protein expression of FOXP1 in Huh7 and SK-HEP-1 cells measured by western blot. (D) Immunohistochemical analysis of FOXP1 in HCC tissues and adjacent noncancer tissues. (E) The mRNA expression levels of FOXP1 in HCC tissues and adjacent noncancer tissues measured by qRT-PCR ( N = 87). (F) Negative correlation between miR-338-3p and FOXP1 in HCC patients. Positive correlation between hsa_circ_104566 and FOXP1 in HCC patients. (G) Protein expression levels of SOX12 in clear cell renal cell carcinoma tissues and adjacent noncancer tissues measured by western blot. FOXP1: forkhead box protein 1; HCC: hepatocellular carcinoma; qRT-PCR: quantitative real-time polymerase chain reaction.

Article Snippet: HCC cell lines (SK-HEP-1, HLE, SNU449, Hep-3B, Huh7) and MIHA normal liver cell line, obtained from the Chinese Academy of Sciences (Shanghai, China), were cultured in Roswell Park Memorial Institute Medium (RPMI) 1640 medium (Lonza, Basel, Switzerland) supplemented with 10% fetal bovine serum (FBS; Gibco, Waltham, MA, USA) at 37 °C constant temperature and incubated with 5% carbon dioxide.

Techniques: Binding Assay, Luciferase, Expressing, Western Blot, Immunohistochemical staining, Quantitative RT-PCR, Real-time Polymerase Chain Reaction

Journal: Cell Transplantation

Article Title: circRNA hsa_circ_104566 Sponged miR-338-3p to Promote Hepatocellular Carcinoma Progression

doi: 10.1177/0963689720963948

Figure Lengend Snippet: Hsa_circ_104566 promoted malignant behaviors of HCC through miR-338-3p/FOXP1. (A) The influence of hsa_circ_104566 and miR-338-3p on protein expression of FOXP1 in Huh7 and SK-HEP-1 cells measured by western blot. (B) The influence of hsa_circ_104566 and FOXP1 on cell viability of Huh7 and SK-HEP-1 cells measured by cell counting kit-8. (C) The influence of hsa_circ_104566 and FOXP1 on cell proliferation of Huh7 and SK-HEP-1 cells measured by the EdU staining assay. (D) The influence of hsa_circ_104566 and FOXP1 on cell apoptosis of Huh7 and SK-HEP-1 cells measured by flow cytometry. (E) The influence of hsa_circ_104566 and FOXP1 on cell migration of Huh7 and SK-HEP-1 cells measured by wound healing assay. (F) The influence of hsa_circ_104566 and FOXP1 on cell invasion of Huh7 and SK-HEP-1 cells measured by Transwell assay. (G) The influence of hsa_circ_104566 and FOXP1 on protein expression levels of FOXP1, PCNA, c-Myc, Bcl-2, cleaved caspase-3, E-cadherin, and N-cadherin in Huh7 and SK-HEP-1 cells measured by western blot. FOXP1: forkhead box protein 1; HCC: hepatocellular carcinoma; PCNA: proliferating cell nuclear antigen.

Article Snippet: HCC cell lines (SK-HEP-1, HLE, SNU449, Hep-3B, Huh7) and MIHA normal liver cell line, obtained from the Chinese Academy of Sciences (Shanghai, China), were cultured in Roswell Park Memorial Institute Medium (RPMI) 1640 medium (Lonza, Basel, Switzerland) supplemented with 10% fetal bovine serum (FBS; Gibco, Waltham, MA, USA) at 37 °C constant temperature and incubated with 5% carbon dioxide.

Techniques: Expressing, Western Blot, Cell Counting, Staining, Flow Cytometry, Migration, Wound Healing Assay, Transwell Assay

Journal: Journal of Gastrointestinal Oncology

Article Title: The deficiency of FKBP-5 inhibited hepatocellular progression by increasing the infiltration of distinct immune cells and inhibiting obesity-associated gut microbial metabolite

doi: 10.21037/jgo-21-71

Figure Lengend Snippet: FKBP-5 was highly expressed in HCC. (A) The expression of FKBP-5 was examined by the IHC staining of para-carcinoma tissues and HCC tissues. The representative images were displayed. Magnification, 100×; Scale bar =100 µm. (B) The western blotting analysis of FKBP-5 expression in HCC tissues (He) and para-carcinoma tissues (Pa), GAPDH was used as an internal reference. (C) The mRNA expression level of FKBP-5 was confirmed by RT-qPCR assay in HL-7702 and five HCC cells (SMMC-7721, Hep 3B, Huh7, Hep G2, and LO2), *, P<0.05; ***, P<0.001 vs. HL-7702 cells.

Article Snippet: Cell culture Human liver (HL)-7702 and HCC cells (i.e., SMMC-7721, Hep 3B, Huh7, Hep G2, and LO2) were purchased from the American Type Culture Collection.

Techniques: Expressing, Immunohistochemistry, Western Blot, Quantitative RT-PCR

Journal: OncoTargets and therapy

Article Title: circ_0003418 Inhibits Tumorigenesis And Cisplatin Chemoresistance Through Wnt/β-Catenin Pathway In Hepatocellular Carcinoma

doi: 10.2147/OTT.S229507

Figure Lengend Snippet: circ_0003418 downregulation in HCC tissues and cells. Notes: ( A ) The expression profile of circ_0003418 in HCC tissues and adjacent noncancerous tissues were detected by qRT-PCR assays. ( B ) qRT-PCR was used to analyze the level of circ_0003418 in five HCC cell lines and one normal human hepatocyte cell line. ( C and D ) The knockdown efficiency of LV3-circ_0003418 on circ_0003418 in Huh-7 and Hep-3B cells was verified via qRT-PCR. ** P<0.01 and *** P<0.001 compared to control group. Abbreviations: HCC, hepatocellular carcinoma; NC, negative control.

Article Snippet: Human HCC cell lines (Hep-3B, Huh-7, Sk-hep-1, SMMC-7721 and PLC) and normal human hepatocyte line (HL-77O2) were purchased from the China Center for Type Culture Collection (Wuhan, China).

Techniques: Expressing, Quantitative RT-PCR, Knockdown, Control, Negative Control

Journal: OncoTargets and therapy

Article Title: circ_0003418 Inhibits Tumorigenesis And Cisplatin Chemoresistance Through Wnt/β-Catenin Pathway In Hepatocellular Carcinoma

doi: 10.2147/OTT.S229507

Figure Lengend Snippet: circ_0003418 suppresses proliferation, migration, and invasion and promotes apoptosis in HCC cells. Notes: ( A and B ) CCK-8 assays were performed to measure the effect of silencing circ_0003418 on the proliferation in Huh-7 and Hep-3B cells. ( C–F ) Effect of silencing circ_0003418 on cell migration ( C and D ) and invasion ( E and F ) were analyzed by transwell migration and invasion assays, respectively. * P<0.05, ** P<0.01 and *** P<0.001 compared to control group. Abbreviations: CCK-8, cell counting kit 8; NC, negative control.

Article Snippet: Human HCC cell lines (Hep-3B, Huh-7, Sk-hep-1, SMMC-7721 and PLC) and normal human hepatocyte line (HL-77O2) were purchased from the China Center for Type Culture Collection (Wuhan, China).

Techniques: Migration, CCK-8 Assay, Control, Cell Counting, Negative Control

Journal: OncoTargets and therapy

Article Title: circ_0003418 Inhibits Tumorigenesis And Cisplatin Chemoresistance Through Wnt/β-Catenin Pathway In Hepatocellular Carcinoma

doi: 10.2147/OTT.S229507

Figure Lengend Snippet: circ-0003418 sensitizes HCC cells to cisplatin in vitro. Notes: ( A and B ) Huh-7 and Hep-3B cells infected with LV3-NC or LV3-circ_0003418 were treated with different doses of cisplatin (1, 2, 4, 8, 16, 32, 64 and 128 mg/L) for 24 hrs, and then cell viability was determined by CCK-8 assays. ( C–F ) Huh-7 cells were treated with cisplatin (11.39 mg/L) for 24 hrs as well as Hep-3B cells were treated with cisplatin (20.18 mg/L) for 24 hrs, and then cell migration and invasion were detected by transwell migration ( C and D ) and invasion ( E and F ) assays, respectively. * P<0.05, ** P<0.01 and *** P<0.001 compared to control group. Abbreviation: NC, negative control.

Article Snippet: Human HCC cell lines (Hep-3B, Huh-7, Sk-hep-1, SMMC-7721 and PLC) and normal human hepatocyte line (HL-77O2) were purchased from the China Center for Type Culture Collection (Wuhan, China).

Techniques: In Vitro, Infection, CCK-8 Assay, Migration, Control, Negative Control

Journal: OncoTargets and therapy

Article Title: circ_0003418 Inhibits Tumorigenesis And Cisplatin Chemoresistance Through Wnt/β-Catenin Pathway In Hepatocellular Carcinoma

doi: 10.2147/OTT.S229507

Figure Lengend Snippet: circ-0003418 enhances sensitivity of HCC cells to cisplatin in vivo. Female BALB/c nude mice were implanted subcutaneously with Huh-7 cells infected with LV3-NC or LV3-circ_0003418. Twelve days later, the LV3-NC tumor-bearing mice were treated with saline or cisplatin (5 mg/kg) by intraperitoneal injection twice a week up to 36 days posttreatment. The mice bearing LV3-circ_0003418 tumors received the same treatment. Notes: ( A ) Image of the tumors in the nude mice. ( B ) The tumors growth curve of xenograft model mice. ( C and D ) The weight and volume of the tumors in the nude mice. * P<0.05, ** P<0.01 and *** P<0.001 compared to control group. Abbreviation: NC, negative control.

Article Snippet: Human HCC cell lines (Hep-3B, Huh-7, Sk-hep-1, SMMC-7721 and PLC) and normal human hepatocyte line (HL-77O2) were purchased from the China Center for Type Culture Collection (Wuhan, China).

Techniques: In Vivo, Infection, Saline, Injection, Control, Negative Control

Journal: OncoTargets and therapy

Article Title: circ_0003418 Inhibits Tumorigenesis And Cisplatin Chemoresistance Through Wnt/β-Catenin Pathway In Hepatocellular Carcinoma

doi: 10.2147/OTT.S229507

Figure Lengend Snippet: Silencing circ_0003418 induces cisplatin resistance of HCC cells through activating the Wnt/β-catenin pathway. Notes: ( A and B ) Huh-7 cells were treated with cisplatin (11.39 mg/L) for 24 hrs as well as Hep-3B cells were treated with cisplatin (20.18 mg/L) for 24 hrs, and then the protein levels of β-catenin and c-Myc in the Huh-7 and Hep-3B cells were detected by Western blotting. ( C and D ) The inhibition efficiency of ICG-001 was detected by Western blotting. ( E and F ) Cell proliferation assay showed that inhibition of Wnt/β-catenin pathway in cells infected with LV3-circ_0003418 inhibited cell proliferation. ( G and H ) Chemotherapy sensitivity assay showed that inhibition of Wnt/β-catenin pathway in cells infected with LV3-circ_0003418 enhanced sensitivity of HCC cells to cisplatin. * P<0.05, ** P<0.01 and *** P<0.001 compared to control group. Abbreviation: NC, negative control.

Article Snippet: Human HCC cell lines (Hep-3B, Huh-7, Sk-hep-1, SMMC-7721 and PLC) and normal human hepatocyte line (HL-77O2) were purchased from the China Center for Type Culture Collection (Wuhan, China).

Techniques: Western Blot, Inhibition, Proliferation Assay, Infection, Sensitive Assay, Control, Negative Control

Journal: Molecular Therapy. Nucleic Acids

Article Title: lncRNA PRR34-AS1 promotes HCC development via modulating Wnt/β-catenin pathway by absorbing miR-296-5p and upregulating E2F2 and SOX12

doi: 10.1016/j.omtn.2021.04.016

Figure Lengend Snippet: PRR34-AS1 expression is elevated in HCC cells (A) UCSC database presented PRR34-AS1 expression profile in multiple human normal tissues, including liver tissues. (B) NONCODE database indicated the profile of PRR34-AS1 expression in different normal tissue samples. (C) GEPIA2 database exhibited PRR34-AS1 expression pattern in 369 cases of LIHC tissues and 160 cases of normal tissues. (D) Quantitative real-time RT-PCR analyzed PRR34-AS1 expression in HCC cell lines (Hep 3B, SK-HEP-1, Huh7, MHCC97-H, and HCCLM3) and human normal hepatocyte cell line (THLE-3). (E and F) Subcellular fractionation and FISH experiments determined PRR34-AS1 location in HCC cells. (G) Knockdown efficiencies of sh-PRR34-AS1#1&2&3 in MHCC97-H and HCCLM3 cells, as well as overexpression efficiency of pcDNA3.1/PRR34-AS1 in Hep 3B cells, were evaluated via quantitative real-time RT-PCR. ∗p < 0.05, ∗∗p < 0.01.

Article Snippet: Human HCC cell lines (Hep 3B and SK-HEP-1), human normal hepatocyte cell line (THLE-3), and human embryonic kidney cell line (HEK293T) were acquired from American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Expressing, Quantitative RT-PCR, Fractionation, Knockdown, Over Expression

Journal: Molecular Therapy. Nucleic Acids

Article Title: lncRNA PRR34-AS1 promotes HCC development via modulating Wnt/β-catenin pathway by absorbing miR-296-5p and upregulating E2F2 and SOX12

doi: 10.1016/j.omtn.2021.04.016

Figure Lengend Snippet: PRR34-AS1 facilitates HCC cell proliferation in vitro and tumor growth in vivo (A) CCK-8 assays tested the changes in cell proliferation ability induced by PRR34-AS1 silencing in MHCC97-H and HCCLM3 cells and induced by PRR34-AS1 upregulation in Hep 3B cells. (B) Colony formation assays measured the number of colonies formed under PRR34-AS1 silencing in MHCC97-H, HCCLM3 cells, or under PRR34-AS1 upregulation in Hep 3B cells. (C and D) TUNEL assays and flow cytometry analysis assessed the apoptosis rate when PRR34-AS1 was downregulated in HCCLM3 cells (E) Representative pictures of xenografts derived from HCCLM3 cells transfected with sh-NC or sh-PRR34-AS1#1. (F) The grow curves of tumors from two different groups. (G and H) The final tumor volume and weight were measured. ∗p < 0.05, ∗∗p < 0.01.

Article Snippet: Human HCC cell lines (Hep 3B and SK-HEP-1), human normal hepatocyte cell line (THLE-3), and human embryonic kidney cell line (HEK293T) were acquired from American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: In Vitro, In Vivo, CCK-8 Assay, TUNEL Assay, Flow Cytometry, Derivative Assay, Transfection

Journal: Molecular Therapy. Nucleic Acids

Article Title: lncRNA PRR34-AS1 promotes HCC development via modulating Wnt/β-catenin pathway by absorbing miR-296-5p and upregulating E2F2 and SOX12

doi: 10.1016/j.omtn.2021.04.016

Figure Lengend Snippet: PRR34-AS1 contributes to cell migration, invasion, and EMT process in HCC cells and activates the Wnt/β-catenin pathway (A) Wound healing assays detected the migration ability of indicated HCC cells when PRR34-AS1 was knocked down or upregulated. (B and C) Transwell experiments analyzed the migration and invasion properties in PRR34-AS1 silenced MHCC97-H and HCCLM3 cells, as well as in PRR34-AS1 upregulated Hep 3B cells. (D) Western blot assay examined the protein levels of E-cadherin, N-cadherin, and Vimentin in HCC cells under PRR34-AS1 depletion or overexpression. (E) H&E staining evaluated the metastatic ability of HCCLM3 cells after PRR34-AS1 depletion. (F) TOP Flash/FOP Flash reporter assays assessed the activity of Wnt/β-catenin in HCC cells with PRR34-AS1 depletion or overexpression. (G–I) Western blot evaluated the levels of indicated proteins in HCC cells after PRR34-AS1 depletion or overexpression. ∗p < 0.05.

Article Snippet: Human HCC cell lines (Hep 3B and SK-HEP-1), human normal hepatocyte cell line (THLE-3), and human embryonic kidney cell line (HEK293T) were acquired from American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Migration, Western Blot, Over Expression, Staining, Activity Assay

Journal: Molecular Therapy. Nucleic Acids

Article Title: lncRNA PRR34-AS1 promotes HCC development via modulating Wnt/β-catenin pathway by absorbing miR-296-5p and upregulating E2F2 and SOX12

doi: 10.1016/j.omtn.2021.04.016

Figure Lengend Snippet: PRR34-AS1 sponges miR-296-5p in HCC cells (A) LncBase database was used to predict underlying miRNAs that could combine with PRR34-AS1. (B) RNA pull-down assays assessed the abundance of candidate miRNAs in biotinylated PRR34-AS1 group in Hep 3B cells. (C) ENCORI database predicted the underlying binding sites between PRR34-AS1 and miR-296-5p. (D) Quantitative real-time RT-PCR analyzed miR-296-5p expression pattern in HCC cells (Hep 3B, SK-HEP-1, Huh7, MHCC97-H, and HCCLM3) and human normal hepatocyte cells (THLE-3). (E) RIP assays assessed the abundance of PRR34-AS1 and miR-296-5p in AGO2 complex. (F) Luciferase reporter assays detected the luciferase activity of PRR34-AS1-WT or PRR34-AS1-Mut under miR-296-5p upregulation in HCC cells. ∗p < 0.05, ∗∗p < 0.01.

Article Snippet: Human HCC cell lines (Hep 3B and SK-HEP-1), human normal hepatocyte cell line (THLE-3), and human embryonic kidney cell line (HEK293T) were acquired from American Type Culture Collection (ATCC; Manassas, VA, USA).

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

Journal: Molecular Therapy. Nucleic Acids

Article Title: lncRNA PRR34-AS1 promotes HCC development via modulating Wnt/β-catenin pathway by absorbing miR-296-5p and upregulating E2F2 and SOX12

doi: 10.1016/j.omtn.2021.04.016

Figure Lengend Snippet: PRR34-AS1 aggravates HCC cell proliferation, migration, and EMT through targeting miR-296-5p (A) Quantitative real-time RT-PCR detected the expression of miR-296-5p in HCCLM3 cells under indicated transfections. (B and C) CCK-8 and colony formation assays examined the proliferation ability of indicated HCCLM3 cells. (D and E) TUNEL and flow cytometry experiments analyzed the apoptosis rate of indicated HCCLM3 cells. (F–H) Wound healing and Transwell assays detected the migratory and invasive capacities of indicated HCCLM3 cells. (I) Western blot examined the protein levels of E-cadherin, N-cadherin, and Vimentin in HCCLM3 cells under different conditions. ∗p < 0.05.

Article Snippet: Human HCC cell lines (Hep 3B and SK-HEP-1), human normal hepatocyte cell line (THLE-3), and human embryonic kidney cell line (HEK293T) were acquired from American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Migration, Quantitative RT-PCR, Expressing, Transfection, CCK-8 Assay, TUNEL Assay, Flow Cytometry, Western Blot

Journal: Molecular Therapy. Nucleic Acids

Article Title: lncRNA PRR34-AS1 promotes HCC development via modulating Wnt/β-catenin pathway by absorbing miR-296-5p and upregulating E2F2 and SOX12

doi: 10.1016/j.omtn.2021.04.016

Figure Lengend Snippet: E2F2 and SOX12 are identified as the targets of miR-296-5p in HCC cells (A) The mirDIP, microT, PITA, and miRmap databases were used to screen underlying mRNAs that potentially combined with miR-296-5p. (B) Quantitative real-time RT-PCR analyzed the expression of candidate targets (ZNF76, HMGA1, FAM53B, FGFR3, E2F2, HIPK1, SOX12, CDK16, BMF, and SLC16A3) in PRR34-AS1 silenced HCCLM3 cells. (C) ENCORI predicted the binding sites between E2F2/SOX12 and miR-296-5p. (D) Quantitative real-time RT-PCR detected E2F2 and SOX12 mRNA levels in HCC cells (Hep 3B, SK-HEP-1, Huh7, MHCC97-H, and HCCLM3) and THLE-3 cells. (E) Western blot assay analyzed the protein levels of E2F2 and SOX12 in PRR34-AS1 silenced or overexpressed HCC cells. (F) RIP assays assessed the enrichments of PRR34-AS1, miR-296-5p, and E2F2/SOX12 in anti-AGO2 complex. (G) Luciferase reporter experiments assessed the luciferase activity of E2F2-3′ UTR-WT/Mut and SOX12-WT/Mut in HEK293T cells with different transfections. ∗p < 0.05, ∗∗p < 0.01.

Article Snippet: Human HCC cell lines (Hep 3B and SK-HEP-1), human normal hepatocyte cell line (THLE-3), and human embryonic kidney cell line (HEK293T) were acquired from American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Quantitative RT-PCR, Expressing, Binding Assay, Western Blot, Luciferase, Activity Assay, Transfection

Journal: Molecular Therapy. Nucleic Acids

Article Title: lncRNA PRR34-AS1 promotes HCC development via modulating Wnt/β-catenin pathway by absorbing miR-296-5p and upregulating E2F2 and SOX12

doi: 10.1016/j.omtn.2021.04.016

Figure Lengend Snippet: E2F2 activates PRR34-AS1 transcription in HCC cells (A) Western blot analyzed the knockdown or overexpression efficiencies of E2F2 in HCC cells. (B and C) The levels of E2F2 and PRR34-AS1 in indicated HCC cells were examined via quantitative real-time RT-PCR. (D) ChIP experiments measured the connection between PRR34-AS1 promoter and E2F2 in HCC cells. (E) The binding motif of E2F2 were predicted by JASPAR. (F) Luciferase reporter experiments assessed the luciferase activities of PRR34-AS1-Pro-WT and PRR34-AS1-Pro-Mut in HCC cells after E2F2 expression was increased or silenced. ∗∗p < 0.01.

Article Snippet: Human HCC cell lines (Hep 3B and SK-HEP-1), human normal hepatocyte cell line (THLE-3), and human embryonic kidney cell line (HEK293T) were acquired from American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Western Blot, Knockdown, Over Expression, Quantitative RT-PCR, Binding Assay, Luciferase, Expressing

Journal: Disease markers

Article Title: hsa_circ_0129047 Upregulates LYVE1 to Inhibit Hepatocellular Carcinoma Progression by Sponging miR-492.

doi: 10.1155/2023/6978234

Figure Lengend Snippet: FIGURE 2: circ_0129047 overexpression delays in vitro and in vivo HCC growth. Overexpression of circ_0129047 (OE-circ) and empty vectors (OE-NC) was induced in Hep 3B and Huh7 cells. (a) RT-qPCR was verifying the exogenous expression of circ_0129047 in Hep 3B and Huh7 cells. (b) CCK8 assays examining the proliferation of Hep 3B and Huh7 cells transfected with OE-circ and OE-NC. (c) Scratch migration assay for examining HCC cell migration. (d) HCC cell invasion ability was detected using a transwell invasion assay. (e) Hep 3B cells overexpressing circ_0129047 or NC were introduced into nude mice through subcutaneous flank injection. Tumor volumes were recorded every 7 days, and xenograft tumors were excised and weighed at 5 weeks. The xenograft tumors were photographed, measured, and weighed. ∗∗P<0:001, vs. OE-NC.

Article Snippet: A human liver cell line (THLE2) and two HCC cell lines (Hep 3B and SNU-182) were purchased from the American Type Culture Collection (ATCC), Manassas, VA, USA.

Techniques: Over Expression, In Vitro, In Vivo, Quantitative RT-PCR, Expressing, Transfection, Migration, Transwell Invasion Assay, Injection

Journal: Disease markers

Article Title: hsa_circ_0129047 Upregulates LYVE1 to Inhibit Hepatocellular Carcinoma Progression by Sponging miR-492.

doi: 10.1155/2023/6978234

Figure Lengend Snippet: FIGURE 4: miR-492 downregulation is critical for the inhibitory effect of circ_0129047 on HCC cell survival. Hep 3B and Huh7 cells were transfected with OE-NC, OE-circ (OE-circ_0129047), miR-492 mimic, NC mimic, or OE-circ+mimic. (a) CCK8 assays were conducted to determine HCC cell proliferation. (b) Scratch migration assays were conducted to evaluate the migration rate of HCC cells after 24 hr of transfection. (c) Transwell migration assays were conducted to examine HCC cell invasion after 48 hr of transfection. ∗P<0:05; ∗∗P<0:001, vs. mimic; ##P<0:001, vs.mimic-NC; &P<0:05; &&P<0:001, vs.OE + mimic.

Article Snippet: A human liver cell line (THLE2) and two HCC cell lines (Hep 3B and SNU-182) were purchased from the American Type Culture Collection (ATCC), Manassas, VA, USA.

Techniques: Transfection, Migration

Journal: Disease markers

Article Title: hsa_circ_0129047 Upregulates LYVE1 to Inhibit Hepatocellular Carcinoma Progression by Sponging miR-492.

doi: 10.1155/2023/6978234

Figure Lengend Snippet: FIGURE 5: MiR-492 targets LYVE1. (a) TargetScan was used to identify the target of miR-492. (b) Luciferase activity driven by 3′UTR LYVE1 MUT and WT was assessed in Hep 3B and Huh7 cells cotransfected with miR-492 mimic or mimic NC. ∗∗P<0:001, vs. miR-NC. (c) RT- qPCR analysis of LYVE1 expression in HCC and normal tissues. (d) RT-qPCR analysis of LYVE1 in THLE2 and HCC cells. ∗∗P<0:001, vs. THLE2. (e) Pearson analysis of the association between LYVE1 and miR-492 in HCC tissues. (f) Western blotting was conducted to examine the LYVE1 expression in Hep 3B and Huh7 cells transfected with OE-NC, mimic NC, OE-circ mimic, or OE-circ + mimic. ∗∗P<0:001, vs. OE-NC; ##P<0:001, vs.mimic-NC; &&P<0:001, vs.OE- circ + mimic.

Article Snippet: A human liver cell line (THLE2) and two HCC cell lines (Hep 3B and SNU-182) were purchased from the American Type Culture Collection (ATCC), Manassas, VA, USA.

Techniques: Luciferase, Activity Assay, Quantitative RT-PCR, Expressing, Western Blot, Transfection

Journal: Disease markers

Article Title: hsa_circ_0129047 Upregulates LYVE1 to Inhibit Hepatocellular Carcinoma Progression by Sponging miR-492.

doi: 10.1155/2023/6978234

Figure Lengend Snippet: FIGURE 6: LYVE1 downregulation contributes to the oncogenic behaviors of miR-492. Hep 3B and Huh7 cells were transfected with OE-NC, mimic NC, OE-LYVE1 mimic, or OE-LYVE1+mimic. (a) Western blot conducted to examine LYVE1 expression in Hep 3B and Huh7 cells. (b) The CCK8 assay was conducted to determine HCC cell proliferation. (c) Scratch migration assays were conducted to evaluate the migration rate of HCC cells after 24 hr of transfection. (d) Transwell migration assays were conducted to examine HCC cell invasion after 48 hr of transfection. ∗∗P<0:001, vs.OE-NC; #P<0:05; ##P<0:001, vs. mimic-NC; &P<0:05; &&P<0:001, vs. OE + mimic.

Article Snippet: A human liver cell line (THLE2) and two HCC cell lines (Hep 3B and SNU-182) were purchased from the American Type Culture Collection (ATCC), Manassas, VA, USA.

Techniques: Transfection, Western Blot, Expressing, CCK-8 Assay, Migration

Journal: Discover Oncology

Article Title: BIRC5 knockdown ameliorates hepatocellular carcinoma progression via regulating PPARγ pathway and cuproptosis

doi: 10.1007/s12672-024-01592-y

Figure Lengend Snippet: BIRC5 knockdown suppresses proliferation, invasion, migration and promotes apoptosis of HCC cells. A QRT-PCR assay was used to verify the knockdown efficiency of BIRC5 expression in both SK-Hep-1 and Hep-3B cells. B Cell viability of SK-Hep-1 and Hep-3B cells with or without BIRC5 knockdown was detected by CCK-8 assay. C Cell migration in si-NC and si-BIRC5 groups was measured by scratch assay. Scale bar: 100 μm. D The transwell assay was utilized to detect cell invasion in si-NC and si-BIRC5 groups. Scale bar: 100 μm. E Flow cytometry was performed to detect cell apoptosis of BIRC5 silencing SK-Hep-1 and Hep-3B cells. ** p < 0.01, *** p < 0.001 vs. si-NC

Article Snippet: The normal human liver cell line (L-O2) and human HCC lines (Hep-3B and SK-Hep-1) were obtained from iCell (Shanghai, China), then were cultured in DMEM medium (Hyclone, Logan, UT, USA) containing 10% heat-inactivated FBS, 100 U/mL penicillin and 100 μg/mL streptomycin (Hyclone, Logan, UT, USA) at 37 °C in a humidified atmosphere of 5% CO 2 concentration conditions.

Techniques: Knockdown, Migration, Quantitative RT-PCR, Expressing, CCK-8 Assay, Wound Healing Assay, Transwell Assay, Flow Cytometry

Journal: Discover Oncology

Article Title: BIRC5 knockdown ameliorates hepatocellular carcinoma progression via regulating PPARγ pathway and cuproptosis

doi: 10.1007/s12672-024-01592-y

Figure Lengend Snippet: Knockdown of BIRC5 affects the cuproptosis of HCC cells. A The supernatant contents of pyruvic acid and α-ketoglutaric acid in BIRC5-silenced SK-Hep-1 and Hep-3B cells were detected using assay kits. B The cellular copper ion (Cu 2+ ) level in SK-Hep-1 and Hep-3B cells characterized by knockdown of BIRC5 was measured using inductively coupled plasma-Optical Emission Spectrometer (ICP-OES). C Expressions of FDX1 and DLAT in BIRC5-silenced SK-Hep-1 and Hep-3B cells were detected by Western blot. ** p < 0.01, *** p < 0.001 vs. si-NC

Article Snippet: The normal human liver cell line (L-O2) and human HCC lines (Hep-3B and SK-Hep-1) were obtained from iCell (Shanghai, China), then were cultured in DMEM medium (Hyclone, Logan, UT, USA) containing 10% heat-inactivated FBS, 100 U/mL penicillin and 100 μg/mL streptomycin (Hyclone, Logan, UT, USA) at 37 °C in a humidified atmosphere of 5% CO 2 concentration conditions.

Techniques: Knockdown, Clinical Proteomics, Western Blot

Journal: Discover Oncology

Article Title: BIRC5 knockdown ameliorates hepatocellular carcinoma progression via regulating PPARγ pathway and cuproptosis

doi: 10.1007/s12672-024-01592-y

Figure Lengend Snippet: BIRC5 regulates the PPAR pathway in HCC cells. Protein levels of PPAR-γ and FABPP5 were verified by western blot in SK-Hep-1 and Hep-3B cells between si-NC, si-BIRC5 group. * p < 0.05, *** p < 0.001 vs. si-NC

Article Snippet: The normal human liver cell line (L-O2) and human HCC lines (Hep-3B and SK-Hep-1) were obtained from iCell (Shanghai, China), then were cultured in DMEM medium (Hyclone, Logan, UT, USA) containing 10% heat-inactivated FBS, 100 U/mL penicillin and 100 μg/mL streptomycin (Hyclone, Logan, UT, USA) at 37 °C in a humidified atmosphere of 5% CO 2 concentration conditions.

Techniques: Western Blot