Journal: Translational Oncology

Article Title: A monoclonal antibody targeting B7-H3 in human embryonic stem cells exhibits anti-tumor activity in hepatocellular carcinoma xenografts

doi: 10.1016/j.tranon.2025.102638

Figure Lengend Snippet: Binding profiles of NPB40 on primary cells and various cancer cells. A, Binding profiles of NPB40 were examined by flow cytometry in naïve and primed hESCs, mouse embryonic fibroblasts (MEFs), peripheral blood mononuclear cells (PBMCs), normal hepatocytes and various cancer cell lines, including hepatocellular carcinoma cells (Huh7, HepG2, SNU387, SNU449). B, Immunocytochemical analysis of 2102Ep, Huh7 and SNU449 cells with NPB40. The scale bar is 20μ m.

Article Snippet: Human cancer cell lines Huh7, SNU387 and SNU449 were purchased from the Korean Cell Line Bank (KCLB, Seoul, Korea) and cultured in RPMI-1640 (Biowest, Kansas, MO, USA) medium supplemented with 10 % FBS (Fetal Bovine Serum, Corning, Seoul, Korea) and antibiotic-antimycotic solution (AA, Biowest).

Techniques: Binding Assay, Flow Cytometry

Journal: Journal of Oncology

Article Title: The lncRNA KIF9-AS1 Accelerates Hepatocellular Carcinoma Growth by Recruiting DNMT1 to Promote RAI2 DNA Methylation

doi: 10.1155/2022/3888798

Figure Lengend Snippet: Knocking down lncRNA KIF9-AS1 expression inhibited the proliferation and migration and promoted the apoptosis of HCC cells. Huh-7 cells were transfected with short hairpin (sh)-NC and sh-KIF9-AS1. (a) StarBase was used to predict lncRNA KIF9-AS1 expression in HCC and paracancerous tissues. (b) lncRNA KIF9-AS1 expression in normal liver cells (HHL-5 cells) and HCC cells (Huh-7, BEL-7405, SNU-398, SNU-387, and Li-7 cells) was detected by qRT–PCR. (c) The proliferative capacity of Huh-7 cells was determined by Cell Counting Kit-8 (CCK-8) assay. (d) The migratory ability of Huh-7 cells was analyzed with a scratch assay. (e) The Huh-7 cell apoptosis rate was measured by flow cytometry. (f) Western blotting was performed to detect Bax, Bcl-2, ERK, and pERK expression. The data are expressed as the means ± SD. ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001.

Article Snippet: Normal human hepatic cells (HHL-5 cells) and HCC cell lines (Huh-7, BEL-7405, SNU-398, SNU-387, and Li-7 cell lines) were provided by American Type Culture Collection (ATCC, Virginia, USA).

Techniques: Expressing, Migration, Transfection, Quantitative RT-PCR, Cell Counting, CCK-8 Assay, Wound Healing Assay, Flow Cytometry, Western Blot

Journal: Journal of Oncology

Article Title: The lncRNA KIF9-AS1 Accelerates Hepatocellular Carcinoma Growth by Recruiting DNMT1 to Promote RAI2 DNA Methylation

doi: 10.1155/2022/3888798

Figure Lengend Snippet: Overexpression of RAI2 inhibited the proliferation and migration and promoted the apoptosis of HCC cells. Huh-7 cells were transfected with overexpressing (oe)-NC and oe-RAI2 vectors. (a) qRT–PCR was performed to detect RAI2 expression in normal liver cells (HHL-5 cells) and HCC cells (Huh-7, BEL-7405, SNU-398, SNU-387, and Li-7 cells). (b) The expression of RAI2 was detected by western blotting (WB). (c) A Cell Counting Kit-8 (CCK-8) assay was performed to assess Huh-7-cellcell proliferation. (d) A scratch assay was performed to determine the migratory ability of Huh-7 cells. (e) The Huh-7 cell apoptosis rate was determined by flow cytometry. (f) Bax, Bcl-2, ERK, and pERK expression were measured by WB. The data are expressed as the mean ± SD. ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001.

Article Snippet: Normal human hepatic cells (HHL-5 cells) and HCC cell lines (Huh-7, BEL-7405, SNU-398, SNU-387, and Li-7 cell lines) were provided by American Type Culture Collection (ATCC, Virginia, USA).

Techniques: Over Expression, Migration, Transfection, Quantitative RT-PCR, Expressing, Western Blot, Cell Counting, CCK-8 Assay, Wound Healing Assay, Flow Cytometry

Journal: Journal of Oncology

Article Title: The lncRNA KIF9-AS1 Accelerates Hepatocellular Carcinoma Growth by Recruiting DNMT1 to Promote RAI2 DNA Methylation

doi: 10.1155/2022/3888798

Figure Lengend Snippet: Knocking down RAI2 expression reversed lncRNA KIF9-AS1-knockdown effects on the proliferation, migration, and apoptosis of HCC cells. Huh-7 cells were transfected with short hairpin (sh)-NC, sh − RAI2, sh − KIF9 − AS1 + sh − NC, and sh − KIF9 − AS1 + sh − RAI2. (a) The expression of KIF9-AS1 and RAI2 was detected by qRT–PCR. (b) The expression of RAI2 was measured by western blotting (WB). (c) A Cell Counting Kit-8 (CCK-8) assay revealed the Huh-7-cell proliferative ability. (d) The migratory ability of Huh-7 cells was determined with a scratch assay. (e) The Huh-7 cell apoptosis rate was analyzed by flow cytometry. (f) WB was performed to detect Bax, Bcl-2, ERK, and pERK expression. The data are expressed as the means ± SD. ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001.

Article Snippet: Normal human hepatic cells (HHL-5 cells) and HCC cell lines (Huh-7, BEL-7405, SNU-398, SNU-387, and Li-7 cell lines) were provided by American Type Culture Collection (ATCC, Virginia, USA).

Techniques: Expressing, Knockdown, Migration, Transfection, Quantitative RT-PCR, Western Blot, Cell Counting, CCK-8 Assay, Wound Healing Assay, Flow Cytometry

Journal: Digestive and liver disease : official journal of the Italian Society of Gastroenterology and the Italian Association for the Study of the Liver

Article Title: LncRNA ILF3-AS1 promotes cell migration, invasion and EMT process in hepatocellular carcinoma via the miR-628-5p/MEIS2 axis to activate the Notch pathway.

doi: 10.1016/j.dld.2021.04.036

Figure Lengend Snippet: Fig. 1. Silenced ILF3-AS1 inhibits cell proliferation, migration, invasion and EMT process in hepatocellular carcinoma. (A) RT-qPCR analysis was used to detect the expression of ILF3-AS1 in hepatocellular carcinoma cell lines (SNU-182, SNU-387, Hep3B, Huh-7) and the normal hepatic cell line HH29. (B) The transfection efficiency of sh-ILF3-AS1#1/#2 in SNU-182 and SNU-387 cells was measured by RT-qPCR analysis. (C) The cell viability of SNU-182 and SNU-387 cells transfected with sh-ILF3-AS1#1/#2 was detected using MTT assay. (D-F) The migration and invasion of SNU-182 and SNU-387 cells with silenced ILF3-AS1 was detected by wound healing and Transwell assays. (G) Western blot analysis showed the levels of key proteins (E-cadherin, N-cadherin, Twist1) related to EMT process in SNU-182 and SNU-387 cells after the transfection of sh-ILF3-AS1#1/#2. (H) Western blot was used to detect the protein expression of matrix metalloproteinases (MMP-2, MMP-9) after ILF3-AS1 knockdown in SNU-182 and SNU-387 cells. (I) The nuclear translocation of β-catenin was assessed by the immunofluorescence staining assay. ∗∗∗p < 0.001.

Article Snippet: Cell culture Hepatocellular carcinoma cell lines (SNU-182, catalogue numer: CRL-2235, ATCC; SNU-387, catalogue number: CRL-2237, ATCC; ep3B, catalogue number: HB-8064, ATCC; Huh-7, JCRB0403, JCRB) nd a normal hepatic cell line HH29 were supplied by American ype culture collection (ATCC) and Japanese Collection of Research ioresources Cell Bank (JCRB).

Techniques: Migration, Quantitative RT-PCR, Expressing, Transfection, MTT Assay, Western Blot, Knockdown, Translocation Assay, Staining

Journal: Digestive and liver disease : official journal of the Italian Society of Gastroenterology and the Italian Association for the Study of the Liver

Article Title: LncRNA ILF3-AS1 promotes cell migration, invasion and EMT process in hepatocellular carcinoma via the miR-628-5p/MEIS2 axis to activate the Notch pathway.

doi: 10.1016/j.dld.2021.04.036

Figure Lengend Snippet: Fig. 2. MiR-628–5p is negatively regulated by ILF3-AS1 in hepatocellular carcinoma cells. (A) A FISH assay was performed to detect the subcellular location of ILF3-AS1 in SNU-182 and SNU-387 cells. (B) Subcellular fractionation assay was used to detect the loca- tion of ILF3-AS1 in hepatocellular carcinoma cells. (C) The location of ILF3-AS1 in cells predicted on the Lnclocator website ( http://www.csbio.sjtu.edu.cn/bioinf/lncLocator/ ). (D) The expression of five potential ILF3-AS1-binding miRNAs (miR-628–5p, miR-629–5p, miR-24–3p, miR-493–5p and miR-185–5p) in SNU-182 and SNU-387 cells trans- fected with sh-ILF3-AS1#1were detected by RT-qPCR. (E) The levels of miR-628–5p was assessed by RT-qPCR in hepatocellular carcinoma cell lines than the normal hepatic cell line HH29. (F) Overexpression efficiency of miR-628–5p was detected using RT-qPCR in SNU-182 and SNU-387 cells transfected with miR-628–5p mimics. (G) The binding site between ILF3-AS1 and miR-628–5p. (H) Luciferase reporter assay showed the luciferase activity of ILF3-AS1, and the relationship between ILF3-AS1 and miR-628–5p. (I) RNA pull down assay showed the enrichment of ILF3-AS1 in the complex pulled down by bio-miR-628–5p WT. (J) RIP assay showed the enrichment of ILF3-AS1 and miR-628–5p in the precipitates of anti-Ago2. ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: Cell culture Hepatocellular carcinoma cell lines (SNU-182, catalogue numer: CRL-2235, ATCC; SNU-387, catalogue number: CRL-2237, ATCC; ep3B, catalogue number: HB-8064, ATCC; Huh-7, JCRB0403, JCRB) nd a normal hepatic cell line HH29 were supplied by American ype culture collection (ATCC) and Japanese Collection of Research ioresources Cell Bank (JCRB).

Techniques: Fractionation, Expressing, Binding Assay, Quantitative RT-PCR, Over Expression, Transfection, Luciferase, Reporter Assay, Activity Assay, Pull Down Assay

Journal: Digestive and liver disease : official journal of the Italian Society of Gastroenterology and the Italian Association for the Study of the Liver

Article Title: LncRNA ILF3-AS1 promotes cell migration, invasion and EMT process in hepatocellular carcinoma via the miR-628-5p/MEIS2 axis to activate the Notch pathway.

doi: 10.1016/j.dld.2021.04.036

Figure Lengend Snippet: Fig. 3. MEIS2 is directly targeted by miR-628–5p in hepatocellular carcinoma cells. (A) Levels of MEIS2 in hepatocellular carcinoma were predicted on the starBase. (B) Survival analysis of MEIS2 in liver cancer predicted by the Kaplan-Meier Plotter ( https: //kmplot.com/analysis/index.php?p=service ). (C) RT-qPCR was used to measure the expression of MEIS2 in hepatocellular carcinoma cell lines and normal hepatic cell line HH29. (D-E) The mRNA and protein levels of MEIS2 were detected by RT-qPCR and western blot analyses in hepatocellular carcinoma cells transfected with miR-628–5p mimics or sh-ILF3-AS1#1, sh-ILF3-AS1#1 + miR-628–5p inhibitor. (F) The binding site between miR-628–5p and MEIS2 was presented. (G) Luciferase reporter assay showed the luciferase activities of wild or mutant-type MEIS2 vectors in hepatocellular carcinoma cells transfected with miR-628–5p mimics. (H) RIP assay demonstrated the interaction between MEIS2 and miR-628–5p in hepatocellular carcinoma cells. ∗∗p < 0.01, p ∗∗∗< 0.001.

Article Snippet: Cell culture Hepatocellular carcinoma cell lines (SNU-182, catalogue numer: CRL-2235, ATCC; SNU-387, catalogue number: CRL-2237, ATCC; ep3B, catalogue number: HB-8064, ATCC; Huh-7, JCRB0403, JCRB) nd a normal hepatic cell line HH29 were supplied by American ype culture collection (ATCC) and Japanese Collection of Research ioresources Cell Bank (JCRB).

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

Journal: Digestive and liver disease : official journal of the Italian Society of Gastroenterology and the Italian Association for the Study of the Liver

Article Title: LncRNA ILF3-AS1 promotes cell migration, invasion and EMT process in hepatocellular carcinoma via the miR-628-5p/MEIS2 axis to activate the Notch pathway.

doi: 10.1016/j.dld.2021.04.036

Figure Lengend Snippet: Fig. 4. ILF3-AS1 facilitates the activation of the Notch pathway by upregulating MEIS2 expression in hepatocellular carcinoma cells. (A) The activity of Notch pathway was measured using the RBP-JK luciferase reporter assays in hepatocellular carcinoma cells transfected with sh-ILF3-AS1#1. (B) Western blot was used to assess the levels of proteins on the Jagged-Notch-Slug signaling axis (Jagged 1, Jagged 2, Notch 1, Twist-1, Hes-1, Hey, p65, AKT, EGFR). (C) The expression of p-p65 (Ser276/Ser536) and p-AKT (Ser473, Thr308) was detected at indicated time points (0, 6, 12, 24 h post transfection) using western blot. (D) A colony formation assay was performed to explore the cell proliferation in the sh-ILF3-AS1#1 and sh-ILF3-AS1#1 + rh-NF- κB groups. (E) A wound healing assay was used to assess the cell migration in indicated groups. (F) Western blot revealed the levels of key factors (Notch1, Twist-1) on the Notch pathway in hepatocellular carcinoma cells transfected with miR-628– 5p mimics, miR-628–5p mimics + pcDNA3.1/MEIS2. (G) The expression of key proteins on the Notch pathway was detected by western blot in hepatocellular carcinoma cells transfected with sh-ILF3-AS1#1, sh-ILF3-AS1#1 + miR-628–5p inhibitor, sh-ILF3-AS1#1 + pcDNA3.1/MEIS2. ∗∗p < 0.01, p ∗∗∗< 0.001.

Article Snippet: Cell culture Hepatocellular carcinoma cell lines (SNU-182, catalogue numer: CRL-2235, ATCC; SNU-387, catalogue number: CRL-2237, ATCC; ep3B, catalogue number: HB-8064, ATCC; Huh-7, JCRB0403, JCRB) nd a normal hepatic cell line HH29 were supplied by American ype culture collection (ATCC) and Japanese Collection of Research ioresources Cell Bank (JCRB).

Techniques: Activation Assay, Expressing, Activity Assay, Luciferase, Transfection, Western Blot, Colony Assay, Wound Healing Assay, Migration

Journal: Digestive and liver disease : official journal of the Italian Society of Gastroenterology and the Italian Association for the Study of the Liver

Article Title: LncRNA ILF3-AS1 promotes cell migration, invasion and EMT process in hepatocellular carcinoma via the miR-628-5p/MEIS2 axis to activate the Notch pathway.

doi: 10.1016/j.dld.2021.04.036

Figure Lengend Snippet: Fig. 6. MiR-628–5p suppresses aggressive phenotypes of hepatocellular carcinoma cells by downregulating MEIS2. (A) MTT assay showed the viability of SNU-182 cells transfected with miR-628–5p mimics, miR-628–5p mimics + pcDNA3.1/MEIS2. (B-C) The migration and invasion abilities of SNU-182 cells were assessed by wound healing and Transwell assays in SNU-182 cells transfected with miR-628–5p mimics, miR-628–5p mimics + pcDNA3.1/MEIS2. (D-E) Western blot showed the levels of key proteins on the EMT process in SNU-182 cells after the transfection of miR-628–5p mimics and miR-628–5p mimics + pcDNA3.1/MEIS2. p ∗∗∗< 0.001.

Article Snippet: Cell culture Hepatocellular carcinoma cell lines (SNU-182, catalogue numer: CRL-2235, ATCC; SNU-387, catalogue number: CRL-2237, ATCC; ep3B, catalogue number: HB-8064, ATCC; Huh-7, JCRB0403, JCRB) nd a normal hepatic cell line HH29 were supplied by American ype culture collection (ATCC) and Japanese Collection of Research ioresources Cell Bank (JCRB).

Techniques: MTT Assay, Transfection, Migration, Western Blot

Journal: Digestive and liver disease : official journal of the Italian Society of Gastroenterology and the Italian Association for the Study of the Liver

Article Title: LncRNA ILF3-AS1 promotes cell migration, invasion and EMT process in hepatocellular carcinoma via the miR-628-5p/MEIS2 axis to activate the Notch pathway.

doi: 10.1016/j.dld.2021.04.036

Figure Lengend Snippet: Fig. 5. ILF3-AS1 knockdown inhibits the tumor growth of hepatocellular carcinoma in vivo (A) The tumor growth curve of mice four weeks after injection was presented. (B) The photo of mouse tumor in sh-NC, sh-ILF3-AS1#1, sh-ILF3-AS1#1 + pcDNA3.1/MEIS2 groups. (C) Tumor weight in three groups. p ∗∗∗< 0.001.

Article Snippet: Cell culture Hepatocellular carcinoma cell lines (SNU-182, catalogue numer: CRL-2235, ATCC; SNU-387, catalogue number: CRL-2237, ATCC; ep3B, catalogue number: HB-8064, ATCC; Huh-7, JCRB0403, JCRB) nd a normal hepatic cell line HH29 were supplied by American ype culture collection (ATCC) and Japanese Collection of Research ioresources Cell Bank (JCRB).

Techniques: Knockdown, In Vivo, Injection

Journal: Digestive and liver disease : official journal of the Italian Society of Gastroenterology and the Italian Association for the Study of the Liver

Article Title: LncRNA ILF3-AS1 promotes cell migration, invasion and EMT process in hepatocellular carcinoma via the miR-628-5p/MEIS2 axis to activate the Notch pathway.

doi: 10.1016/j.dld.2021.04.036

Figure Lengend Snippet: Fig. 7. ILF3-AS1 promotes the malignant behaviors of hepatocellular carcinoma cells by upregulating MEIS2 (A) MTT assay showed the viability of SNU-182 cells transfected with sh-ILF3-AS1#1, sh-ILF3-AS1#1 + pcDNA3.1/MEIS2. (B-C) The migration and invasion abilities of SNU- 182 cells were measured using wound healing and Transwell assays in SNU-182 cells transfected with sh-ILF3-AS1#1, sh-ILF3-AS1#1 + pcDNA3.1/MEIS2. (D-E) Western blot showed the levels of key proteins on the EMT process in SNU-182 cells posttransfection of sh-ILF3-AS1#1 and sh-ILF3-AS1#1 + pcDNA3.1/MEIS2. p ∗∗∗< 0.001.

Article Snippet: Cell culture Hepatocellular carcinoma cell lines (SNU-182, catalogue numer: CRL-2235, ATCC; SNU-387, catalogue number: CRL-2237, ATCC; ep3B, catalogue number: HB-8064, ATCC; Huh-7, JCRB0403, JCRB) nd a normal hepatic cell line HH29 were supplied by American ype culture collection (ATCC) and Japanese Collection of Research ioresources Cell Bank (JCRB).

Techniques: MTT Assay, Transfection, Migration, Western Blot