Journal: Oncology Reports

Article Title: Long non-coding RNA H19 is involved in sorafenib resistance in hepatocellular carcinoma by upregulating miR-675

doi: 10.3892/or.2020.7608

Figure Lengend Snippet: H19 expression is negatively related to sorafenib sensitivity in HCC cell linws (Huh7, Hep3B, SNU-449, SNU-387). (A) Cell viability was examined by CCK-8 assay in four HCC cell lines subjected to sorafenib treatment. (B) Statistical analysis of the IC 50 in four HCC cell lines subjected to sorafenib treatment. (C) Relative mRNA levels of H19 as measured by RT-qPCR and normalized to β-actin in four HCC cell lines. (D) Cell proliferation was examined by EdU assay in four HCC cell lines subjected to sorafenib treatment. **P<0.01, ***P<0.001, compared to the control. HCC, hepatocellular carcinoma; IC 50 , half maximal inhibitory concentration.

Article Snippet: Human HCC cell lines (Huh7, Hep3B, SNU-449, SNU-387) were purchased from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Expressing, CCK-8 Assay, Quantitative RT-PCR, EdU Assay, Control, Concentration Assay

Journal: Oncology Reports

Article Title: Long non-coding RNA H19 is involved in sorafenib resistance in hepatocellular carcinoma by upregulating miR-675

doi: 10.3892/or.2020.7608

Figure Lengend Snippet: H19 knockdown sensitizes HCC cells to sorafenib in vitro . (A) Sorafenib sensitivity of HCC cells transfected with NC siRNA or H19 siRNA. (B) Proliferative capacity of HCC cells transfected with NC siRNA or H19 siRNA and subjected to sorafenib treatment. **P<0.01, ***P<0.001, compared with the sorafenib group. (C) H19 expression in HCC cells transfected with NC siRNA or H19 siRNA and subjected to sorafenib treatment. *P<0.05, **P<0.01, ***P<0.001, compared to the control. HCC, hepatocellular carcinoma; NC, negative control.

Article Snippet: Human HCC cell lines (Huh7, Hep3B, SNU-449, SNU-387) were purchased from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Knockdown, In Vitro, Transfection, Expressing, Control, Negative Control

Journal: Oncology Reports

Article Title: Long non-coding RNA H19 is involved in sorafenib resistance in hepatocellular carcinoma by upregulating miR-675

doi: 10.3892/or.2020.7608

Figure Lengend Snippet: H19 knockdown inhibits EMT and H19 positively regulates miR-675. (A) Immunofluorescence staining showed that E-cadherin (green) and vimentin (green) levels were altered after transfection of NC siRNA or H19 siRNA in the SNU-449 and SNU-387 cells. (B) E-cadherin and vimentin as assessed by western blot analysis. GAPDH was used as loading control. (C) Relative mRNA levels of miR-675 were measured by RT-qPCR and normalized to U6 after transfection of NC siRNA or H19 siRNA in four HCC cell lines. *P<0.05, **P<0.01, ***P<0.001, compared to the control. HCC, hepatocellular carcinoma; NC, negative control; EMT, epithelial-mesenchymal transition.

Article Snippet: Human HCC cell lines (Huh7, Hep3B, SNU-449, SNU-387) were purchased from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Knockdown, Immunofluorescence, Staining, Transfection, Western Blot, Control, Quantitative RT-PCR, Negative Control

Journal: Oncology Reports

Article Title: Long non-coding RNA H19 is involved in sorafenib resistance in hepatocellular carcinoma by upregulating miR-675

doi: 10.3892/or.2020.7608

Figure Lengend Snippet: miR-675 regulates sensitivity of HCC cells to sorafenib. (A) Sorafenib sensitivity of HCC cells transfected with NC siRNA, miR-675 mimic, or miR-675 inhibitor. (B) Proliferation of HCC cells transfected with NC siRNA, miR-675 mimic, or miR-675 inhibitor in the presence of sorafenib. ***P<0.001, compared with the sorafenib group. (C) Expression of miR-675 in HCC cells transfected with NC siRNA, miR-675 mimic, or miR-675 inhibitor in the presence of sorafenib. *P<0.05, **P<0.01, ***P<0.001. HCC, hepatocellular carcinoma; NC, negative control.

Article Snippet: Human HCC cell lines (Huh7, Hep3B, SNU-449, SNU-387) were purchased from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Transfection, Expressing, Negative Control

Journal: Oncology Reports

Article Title: Long non-coding RNA H19 is involved in sorafenib resistance in hepatocellular carcinoma by upregulating miR-675

doi: 10.3892/or.2020.7608

Figure Lengend Snippet: miR-675 alters EMT. (A) Immunofluorescence showed that the staining of E-cadherin (green) and vimentin (green) was altered after transfection with the miR-675 mimic, inhibitor, or NC siRNA in SNU-449 and SNU-387 cells. (B) E-cadherin and vimentin levels in HCC cells transfected with NC siRNA, miR-675 mimic, or miR-675 inhibitor in the presence of sorafenib. HCC, hepatocellular carcinoma; NC, negative control; EMT, epithelial-mesenchymal transition.

Article Snippet: Human HCC cell lines (Huh7, Hep3B, SNU-449, SNU-387) were purchased from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Immunofluorescence, Staining, Transfection, Negative Control

Journal: Oncology Reports

Article Title: Long non-coding RNA H19 is involved in sorafenib resistance in hepatocellular carcinoma by upregulating miR-675

doi: 10.3892/or.2020.7608

Figure Lengend Snippet: miR-675 mediates the regulatory effect of lncRNA H19 on sorafenib sensitivity. Sorafenib sensitivity of HCC cells transfected with NC siRNA or H19 siRNA was evaluated by CCK-8 assay in the presence of the miR-675 mimic for 48 h. HCC, hepatocellular carcinoma; NC, negative control.

Article Snippet: Human HCC cell lines (Huh7, Hep3B, SNU-449, SNU-387) were purchased from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Transfection, CCK-8 Assay, Negative Control

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: OncoTargets and therapy

Article Title: Hsa_circ_0006916 Knockdown Represses the Development of Hepatocellular Carcinoma via Modulating miR-599/SRSF2 Axis

doi: 10.2147/OTT.S267471

Figure Lengend Snippet: Circ_0006916 expression in HCC. ( A ) circ_0006916 level in HCC tissues (n = 30) and normal samples. ( B ) circ_0006916 abundance in Huh7, SNU-387 and THLE-2 cells. ( C and D ) circ_0006916 and HOMER1 levels in Huh7 and SNU-387 cells were detected after treatment of RNase R. ( E and F ) circ_0006916 and HOMER1 levels in Huh7 and SNU-387 cells were measured after treatment of Actinomycin D. * P <0.05.

Article Snippet: Human HCC cell lines (Huh7 and SNU-387 cells) were purchased from Procell (Wuhan, China) and grew in DMEM (Thermo Fisher, Waltham, MA, USA) plus 10% fetal bovine serum (Gibco, Gran Island, NY, USA) and 1% penicillin/streptomycin (Beyotime, Shanghai, China).

Techniques: Expressing

Journal: OncoTargets and therapy

Article Title: Hsa_circ_0006916 Knockdown Represses the Development of Hepatocellular Carcinoma via Modulating miR-599/SRSF2 Axis

doi: 10.2147/OTT.S267471

Figure Lengend Snippet: The impact of circ_0006916 on HCC cell progression. ( A and B ) circ_0006916 and HOMER1 levels were detected in Huh7 and SNU-387 cells with transfection of si-NC or si-circ_0006916. Cell viability ( C ), colony formation ( D ), migration and invasion ( E – G ), cycle distribution ( H ) and apoptosis ( I ) were examined in Huh7 and SNU-387 cells with transfection of si-NC or si-circ_0006916. * P <0.05.

Article Snippet: Human HCC cell lines (Huh7 and SNU-387 cells) were purchased from Procell (Wuhan, China) and grew in DMEM (Thermo Fisher, Waltham, MA, USA) plus 10% fetal bovine serum (Gibco, Gran Island, NY, USA) and 1% penicillin/streptomycin (Beyotime, Shanghai, China).

Techniques: Transfection, Migration

Journal: OncoTargets and therapy

Article Title: Hsa_circ_0006916 Knockdown Represses the Development of Hepatocellular Carcinoma via Modulating miR-599/SRSF2 Axis

doi: 10.2147/OTT.S267471

Figure Lengend Snippet: The association of circ_0006916 and miR-599. ( A ) The target sequence of circ_0006916 and miR-599. ( B and C ) Luciferase activity was detected in Huh7 and SNU-387 cells with transfection of circ_0006916 WT or circ_0006916 MUT and miR-599 mimic or miRNA NC. ( D ) miR-599 level in HCC tissue samples (n = 30) and normal samples. ( E ) miR-599 abundance in Huh7, SNU-387 and THLE-2 cells. ( F and G ) circ_0006916 and miR-599 levels were detected in Huh7 and SNU-387 cells after RIP. * P <0.05.

Article Snippet: Human HCC cell lines (Huh7 and SNU-387 cells) were purchased from Procell (Wuhan, China) and grew in DMEM (Thermo Fisher, Waltham, MA, USA) plus 10% fetal bovine serum (Gibco, Gran Island, NY, USA) and 1% penicillin/streptomycin (Beyotime, Shanghai, China).

Techniques: Sequencing, Luciferase, Activity Assay, Transfection

Journal: OncoTargets and therapy

Article Title: Hsa_circ_0006916 Knockdown Represses the Development of Hepatocellular Carcinoma via Modulating miR-599/SRSF2 Axis

doi: 10.2147/OTT.S267471

Figure Lengend Snippet: The effect of miR-599 on circ_0006916-mediated HCC cell progression. ( A ) miR-599 expression was detected in Huh7 and SNU-387 cells with transfection of inhibitor NC or miR-599 inhibitor. miR-599 abundance ( B ), cell viability ( C ), colony formation ( D ), migration and invasion ( E – G ), cycle distribution ( H and I ) and apoptosis ( J ) were measured in Huh7 and SNU-387 cells with transfection of si-NC, si-circ_0006916, si-circ_0006916 + inhibitor NC or miR-599 inhibitor. * P <0.05.

Article Snippet: Human HCC cell lines (Huh7 and SNU-387 cells) were purchased from Procell (Wuhan, China) and grew in DMEM (Thermo Fisher, Waltham, MA, USA) plus 10% fetal bovine serum (Gibco, Gran Island, NY, USA) and 1% penicillin/streptomycin (Beyotime, Shanghai, China).

Techniques: Expressing, Transfection, Migration

Journal: OncoTargets and therapy

Article Title: Hsa_circ_0006916 Knockdown Represses the Development of Hepatocellular Carcinoma via Modulating miR-599/SRSF2 Axis

doi: 10.2147/OTT.S267471

Figure Lengend Snippet: The relationship of miR-599 and SRSF2 . ( A ) The target sequence of miR-599 and SRSF2 . ( B and C ) Luciferase activity was measured in Huh7 and SNU-387 cells with transfection of SRSF2-3ʹUTR WT or SRSF2-3ʹUTR MUT and miR-599 mimic or miRNA NC. ( D and E ) miR-599 and SRSF2 abundances were examined in Huh7 and SNU-387 cells after RIP. ( F ) SRSF2 expression in HCC tissues and normal samples. ( G ) SRSF2 expression in Huh7, SNU-387 and THLE-2 cells. ( H ) SRSF2 abundance was detected in Huh7 and SNU-387 cells with transfection of pc-NC or pc-SRSF2. ( I ) SRSF2 level was detected in cells with transfection of miRNA NC, miR-599 mimic, miR-599 mimic + pc-NC or pc-SRSF2. * P <0.05.

Article Snippet: Human HCC cell lines (Huh7 and SNU-387 cells) were purchased from Procell (Wuhan, China) and grew in DMEM (Thermo Fisher, Waltham, MA, USA) plus 10% fetal bovine serum (Gibco, Gran Island, NY, USA) and 1% penicillin/streptomycin (Beyotime, Shanghai, China).

Techniques: Sequencing, Luciferase, Activity Assay, Transfection, Expressing

Journal: OncoTargets and therapy

Article Title: Hsa_circ_0006916 Knockdown Represses the Development of Hepatocellular Carcinoma via Modulating miR-599/SRSF2 Axis

doi: 10.2147/OTT.S267471

Figure Lengend Snippet: The influence of miR-599 and SRSF2 on HCC cell progression. Cell viability ( A ), colony formation ( B ), migration and invasion ( C – E ), cycle distribution ( F and G ) and apoptosis ( H ) were examined in Huh7 and SNU-387 cells with transfection of miRNA NC, miR-599 mimic, miR-599 mimic + pc-NC or pc-SRSF2. * P <0.05.

Article Snippet: Human HCC cell lines (Huh7 and SNU-387 cells) were purchased from Procell (Wuhan, China) and grew in DMEM (Thermo Fisher, Waltham, MA, USA) plus 10% fetal bovine serum (Gibco, Gran Island, NY, USA) and 1% penicillin/streptomycin (Beyotime, Shanghai, China).

Techniques: Migration, Transfection

Journal: OncoTargets and therapy

Article Title: Hsa_circ_0006916 Knockdown Represses the Development of Hepatocellular Carcinoma via Modulating miR-599/SRSF2 Axis

doi: 10.2147/OTT.S267471

Figure Lengend Snippet: The impact of circ_0006916 on HCC cell growth in vivo. ( A ) Tumor volume was measured every 7 days. ( B ) Tumor weight was examined. ( C – E ) circ_0006916, miR-599 and SRSF2 abundances were examined in tumor tissues. * P <0.05.

Article Snippet: Human HCC cell lines (Huh7 and SNU-387 cells) were purchased from Procell (Wuhan, China) and grew in DMEM (Thermo Fisher, Waltham, MA, USA) plus 10% fetal bovine serum (Gibco, Gran Island, NY, USA) and 1% penicillin/streptomycin (Beyotime, Shanghai, China).

Techniques: In Vivo

Journal: Frontiers in Oncology

Article Title: Metal-Regulatory Transcription Factor-1 Targeted by miR-148a-3p Is Implicated in Human Hepatocellular Carcinoma Progression

doi: 10.3389/fonc.2021.700649

Figure Lengend Snippet: Metal-regulatory transcription factor-1 (MTF-1) promoted cell proliferation, migration, and invasion as a key mediator and regulator. The regulatory roles of MTF-1 in cell proliferation were evaluated using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and colony formation assays (A, B) . Effect of MTF-1 on cell motility was assessed using the wound healing assay. Scale bar = 500 μm (C) . MTF-1-promoted hepatocellular carcinoma (HCC) cell migration and invasion, as determined using the Transwell assay. Scale bar = 500 μm (D, E) . Cell apoptotic rates were detected after different treatments (F) . Co-immunoprecipitation (Co-IP) study showed specific binding between MTF-1 and APE/Ref-1 with different treatments (G) . MTF-1 expression positively correlated with upregulated APE/Ref-1 level, as determined using the integrative analysis of The Cancer Genome Atlas (TCGA) data (H) . Overexpression of MTF-1 activated APE/Ref-1, which in turn initiated the upregulation of MTF-1 level in HCC cells (I, J) . Data are presented as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Human HCC cell lines (Hep3B and SNU-387) were purchased from the Procell Life Science Technology Company (Wuhan, China) with STR certificate.

Techniques: Migration, Wound Healing Assay, Transwell Assay, Immunoprecipitation, Co-Immunoprecipitation Assay, Binding Assay, Expressing, Over Expression

Journal: Frontiers in Oncology

Article Title: Metal-Regulatory Transcription Factor-1 Targeted by miR-148a-3p Is Implicated in Human Hepatocellular Carcinoma Progression

doi: 10.3389/fonc.2021.700649

Figure Lengend Snippet: Metal-regulatory transcription factor-1 (MTF-1) is directly targeted by miR-148a-3p. The predicted binding sites of miR-148a-3p in the 3′-UTR of MTF-1 were determined (A) . Integrative analysis of The Cancer Genome Atlas (TCGA) data indicated that miR-148a-3p expression was downregulated in tumors (B) , and inversely correlated with MTF-1 and APE/Ref-1 levels (C, D) . Strong miR-148a-3p expression was associated with improved survival (E) . MiR-148a-3p level was assessed in tissue microarrays (TMAs) with considerably reduced average positive area in hepatocellular carcinoma (HCC) tissue. Scale bar = 100 μm (F) . Downregulated miR-148a-3p expression was observed in HCC cells (G) . Overexpression of miR-148a-3p reduced luciferase activity of MTF-1-wt (wild-type) instead of MTF-1-mt (mutant-type) (H) . MiR-148a-3p overexpression inhibited MTF-1 expression in HCC cells (I) . Data are expressed as mean ± SD. ** p < 0.01, *** p < 0.001.

Article Snippet: Human HCC cell lines (Hep3B and SNU-387) were purchased from the Procell Life Science Technology Company (Wuhan, China) with STR certificate.

Techniques: Binding Assay, Expressing, Over Expression, Luciferase, Activity Assay, Mutagenesis

Journal: Frontiers in Oncology

Article Title: Metal-Regulatory Transcription Factor-1 Targeted by miR-148a-3p Is Implicated in Human Hepatocellular Carcinoma Progression

doi: 10.3389/fonc.2021.700649

Figure Lengend Snippet: MiR-148a-3p overexpression reversed the effects of activated metal-regulatory transcription factor-1 (MTF-1) in hepatocellular carcinoma (HCC) cells. To further elucidate the regulatory role of miR-148a-3p, Hep3B and SNU-387 cells were treated with or without miR-148a-3p mimics after MTF-1 overexpression. miR-148a-3p overexpression reversed the effects of activated MTF-1 in HCC cell proliferation (A) , colony formation (B) , migration (C) , invasion (D) , and apoptosis (E) . Scale bar = 500 μm. Data are expressed as mean± SD. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Human HCC cell lines (Hep3B and SNU-387) were purchased from the Procell Life Science Technology Company (Wuhan, China) with STR certificate.

Techniques: Over Expression, Migration

Journal: Frontiers in Oncology

Article Title: Metal-Regulatory Transcription Factor-1 Targeted by miR-148a-3p Is Implicated in Human Hepatocellular Carcinoma Progression

doi: 10.3389/fonc.2021.700649

Figure Lengend Snippet: Function of miR-148a-3p on cell viability, colony formation, and metastasis. The effect of miR-148a-3p alone was further evaluated. Capacities of cell proliferation (A) , colony formation (B) , migration (C, D) , and metastasis (E) were suppressed with miR-148a-3p overexpression; miR-148a-3p knockdown facilitated cell mobility and invasion, and enhanced the anti-apoptotic capability (F) in hepatocellular carcinoma (HCC) cells. Scale bar = 500 μm. Data are expressed as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Human HCC cell lines (Hep3B and SNU-387) were purchased from the Procell Life Science Technology Company (Wuhan, China) with STR certificate.

Techniques: Migration, Over Expression, Knockdown

Journal: Frontiers in Oncology

Article Title: Metal-Regulatory Transcription Factor-1 Targeted by miR-148a-3p Is Implicated in Human Hepatocellular Carcinoma Progression

doi: 10.3389/fonc.2021.700649

Figure Lengend Snippet: Inhibitory effects of exosomal miR-148a-3p on hepatocellular carcinoma (HCC) cells. Exosomes were isolated and RNA was extracted. The morphology of exosomes was evaluated by transmission electron microscopy (A) . Size distribution of exosomes was verified (B) . Western blotting was performed to identify exosome markers (C) . Exosomes labeled with PKH-26 red fluorescent dye were observed in Hep3B and SNU-387 cells after co-incubation. Scale bar = 100 μm (D) . miR-148a-3p was downregulated in the exosomes isolated from the plasma of patients with HCC and culture media of HCC cells (E, F) . To further elucidate the role of exosomal miR-148a-3p, miR-148a-3p and metal-regulatory transcription factor-1 (MTF-1) levels were evaluated after culturing with exosomes from hepatocyte- or HCC-conditioned medium (G, H) . HCC cell proliferation, mobility, and invasion were also inhibited in the presence of exosomes isolated from hepatocyte-conditioned medium. Scale bar = 500 μm (I–L) . Co-culture with exosomes from hepatocyte- conditioned medium induced HCC cell apoptosis (M) . Data are expressed as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Human HCC cell lines (Hep3B and SNU-387) were purchased from the Procell Life Science Technology Company (Wuhan, China) with STR certificate.

Techniques: Isolation, Transmission Assay, Electron Microscopy, Western Blot, Labeling, Incubation, Clinical Proteomics, Co-Culture Assay