Journal: Cell Death & Disease

Article Title: Polyphyllin I suppresses the formation of vasculogenic mimicry via Twist1/VE-cadherin pathway

doi: 10.1038/s41419-018-0902-5

Figure Lengend Snippet: a Polyphyllin I exhibited the most obvious inhibitory effect on VM formation. b Structure of PPI. c Effects of PPI on the viability of SMMC7721, PLC, HepG2, Hep3B, and Bel7402 cells. PPI reduced the survival rate of HCC cells at relatively high concentrations and had slight influence on cell survival below 1 μM. d Cell cycle analysis of PPI in HCC cell lines. 0.1 μM PPI did not influence the cycle of HCC cells. e , f PPI inhibited the migration and invasion of HCC cells. g PPI decreased the number of pipe-like structures in SMMC7721 cells on Matrigel. h Time-lapse images of SMMC7721. PPI inhibited the pipe-like structure formation in SMMC7721 cells ( n = 3)

Article Snippet: HCC cell lines including SMMC7721, PLC, HepG2, Hep3B, and Bel7402 were purchased from Keygene BioTECH (Nanjing, China) and validated through a short tandem repeat-based method.

Techniques: Cell Cycle Assay, Migration

Journal: Molecular Medicine Reports

Article Title: Lipid storage droplet protein 5 reduces sodium palmitate-induced lipotoxicity in human normal liver cells by regulating lipid metabolism-related factors

doi: 10.3892/mmr.2019.10360

Figure Lengend Snippet: Sodium palmitate represses the viability of LO2 cells. (A) CCK-8 assay was performed to determine the effects of various concentrations of sodium palmitate (25, 50, 75, 100, 125 and 150 µmol/l) on LO2 human liver cells treated for 12, 24 and 48 h. ^ P<0.05 and ^^ P<0.01 vs. Control. (B) CCK-8 assay was used to measure the effect of LSDP5 on cell viability in LO2 cells treated with 100 µmol/l sodium palmitate for 48 h (Model). *P<0.05 vs. Control; ^ P<0.05 vs. Model. LSDP5, lipid storage droplet protein 5; M, model; NC, negative control.

Article Snippet: LO2 normal human liver cells were purchased from Procell Life Technology Co., Ltd. (Wuhan, China).

Techniques: CCK-8 Assay, Control, Negative Control

Journal: Molecular Medicine Reports

Article Title: Lipid storage droplet protein 5 reduces sodium palmitate-induced lipotoxicity in human normal liver cells by regulating lipid metabolism-related factors

doi: 10.3892/mmr.2019.10360

Figure Lengend Snippet: LSDP5 expression in LO2 cells. (A and B) Experiments were divided into Control group (0.1% PBS treatment), Model group (100 µmol/l sodium palmitate treatment), NC group (Model cells transfected with pCMV5-NC plasmid) and LSDP5 group (Model cells transfected with pCMV5-LSDP5 overexpression plasmid), and the (A) mRNA and (B) protein expression levels were determined reverse transcription-quantitative polymerase chain reaction and western blotting, respectively; β-actin served as an internal control and for normalization. ^ P<0.05 vs. Model. LSDP5, lipid storage droplet protein 5; M, model; NC, negative control.

Article Snippet: LO2 normal human liver cells were purchased from Procell Life Technology Co., Ltd. (Wuhan, China).

Techniques: Expressing, Control, Transfection, Plasmid Preparation, Over Expression, Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot, Negative Control

Journal: Molecular Medicine Reports

Article Title: Lipid storage droplet protein 5 reduces sodium palmitate-induced lipotoxicity in human normal liver cells by regulating lipid metabolism-related factors

doi: 10.3892/mmr.2019.10360

Figure Lengend Snippet: LSDP5 suppresses the activity of oxidative stress in LO2 lipotoxicity Model cells. (A) The rate of ROS production was measured using an ROS detection kit. (B) NEFA content was detected by NEFA detection kit. (C and D) The contents of (C) MDA and (D) SOD were analysed by ELISA. *P<0.05, **P<0.01 and ***P<0.001 vs. Control; ^ P<0.05 and ^^ P<0.01 vs. Model. LSDP5, lipid storage droplet protein 5; M, model; MDA, malondialdehyde; NC, negative control; NEFA, non-esterified fatty acid; SOD, superoxide dismutase.

Article Snippet: LO2 normal human liver cells were purchased from Procell Life Technology Co., Ltd. (Wuhan, China).

Techniques: Activity Assay, Enzyme-linked Immunosorbent Assay, Control, Negative Control

Journal: Molecular Medicine Reports

Article Title: Lipid storage droplet protein 5 reduces sodium palmitate-induced lipotoxicity in human normal liver cells by regulating lipid metabolism-related factors

doi: 10.3892/mmr.2019.10360

Figure Lengend Snippet: LSDP5 reduces apoptosis in LO2 lipotoxicity Model cells. (A) Apoptosis was detected by Annexin V-FITC/PI apoptosis detection kit. (B) mRNA expression levels of Bax and Bcl-2 were measured by reverse transcription-quantitative polymerase chain reaction. (C and D) Protein expression levels of active-caspase-3, Bax and Bcl-2 were measured by western blot assay; β-actin was used as a loading control and for normalization. *P<0.05, **P<0.01 and ***P<0.001 vs. Control; ^ P<0.05 and ^^ P<0.01 vs. Model. Bax, Bcl-2-associated X protein; Bcl-2, B-cell lymphoma-2; FITC, fluorescein isothiocyanate; LSDP5, lipid storage droplet protein 5; M, model; NC, negative control; PI, propidium iodide.

Article Snippet: LO2 normal human liver cells were purchased from Procell Life Technology Co., Ltd. (Wuhan, China).

Techniques: Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot, Control, Negative Control

Journal: Molecular Medicine Reports

Article Title: Lipid storage droplet protein 5 reduces sodium palmitate-induced lipotoxicity in human normal liver cells by regulating lipid metabolism-related factors

doi: 10.3892/mmr.2019.10360

Figure Lengend Snippet: LSDP5 reduces mitochondrial damage in LO2 lipotoxicity Model. (A) MMP rates were detected by JC-1 kit. (B) mRNA expression levels of Cytc, Cox IV and CPT1a were measured by reverse transcription-quantitative polymerase chain reaction. (C and D) Protein expression levels of Cytc, Cox IV and CPT1a were measured by western blot assay; β-actin was used as a loading control and for normalization. *P<0.05 and **P<0.01 vs. Control; ^ P<0.05 vs. Model. Cox IV, cytochrome c oxidase subunit IV; CPT1a, carnitine palmitoyltransferase 1a; Cytc, cytochrome c ; LSDP5, lipid storage droplet protein 5; M, model; MMP, mitochondrial membrane potential; NC, negative control.

Article Snippet: LO2 normal human liver cells were purchased from Procell Life Technology Co., Ltd. (Wuhan, China).

Techniques: Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot, Control, Membrane, Negative Control

Journal: Molecular Medicine Reports

Article Title: Lipid storage droplet protein 5 reduces sodium palmitate-induced lipotoxicity in human normal liver cells by regulating lipid metabolism-related factors

doi: 10.3892/mmr.2019.10360

Figure Lengend Snippet: LSDP5 regulates lipid metabolism-related factors in LO2 lipotoxicity Model cells. (A and B) ACC1, ACC2, Fas and PPARα (A) mRNA and (B) protein expression levels were assessed by reverse transcription-quantitative polymerase chain reaction and western blot analysis, respectively; β-actin served as an internal control and for normalization. *P<0.05 and **P<0.01 vs. Control; ^ P<0.05 and ^^ P<0.01 vs. Model. ACC, acetyl-co A carboxylase1; Fas, fatty acid synthase; LSDP5, lipid storage droplet protein 5; M, model; NC, negative control.

Article Snippet: LO2 normal human liver cells were purchased from Procell Life Technology Co., Ltd. (Wuhan, China).

Techniques: Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot, Control, Negative Control

Journal: Journal of Hepatocellular Carcinoma

Article Title: Coilin Affects the Prognosis of Hepatocellular Carcinoma Through Cell Cycle and Apoptosis

doi: 10.2147/JHC.S500119

Figure Lengend Snippet: Expression of COIL in HCC cell lines and HCC tissue from hospital. The expression of COIL in liver cancer cell lines huh7, hepG2 and hep3B was higher than that in L02 by immunohistochemistry ( A ) and Western blotting ( B ). ( C ) The relative expression of COIL in HCC was significantly higher than that in normal liver tissue by qRT-PCR (***p value < 0.001). ( D ) The expression of COIL in HCC tissue was higher than that in normal liver tissue. ( E, F ) Representative immunohistochemistry results of low or high COIL expression. ( G ) High COIL expression was associated with poor overall survival.

Article Snippet: The human normal liver cell line L02 was obtained from Beijing Baiao Bowei Biotechnology Co., Ltd.

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

Journal: OncoTargets and therapy

Article Title: lncRNA TMPO-AS1 Exerts Oncogenic Roles in HCC Through Regulating miR-320a/SERBP1 Axis

doi: 10.2147/OTT.S250355

Figure Lengend Snippet: High TMPO-AS1 expression is identified in HCC cells and tissues. ( A ) The expression levels of TMPO-AS1 in HCC tissues and non-cancerous tissues were detected using qRT-PCR assay. ** P <0.01 compared with non-cancerous. ( B ) Expression levels of TMPO-AS1 in HCC tissues from stage I–II and stage III–IV. * P <0.01 compared with I–II. ( C ) Expression levels of TMPO-AS1 in HCC patients with metastasis and without metastasis. * P <0.01 compared with no metastasis. ( D ) Kaplan–Meier curves for HCC patients with higher expression of TMPO-AS1 or lower expression of TMPO-AS1. ( E ) The levels of TMPO-AS1 in HCC cell lines and LO2 cell were detected using qRT-PCR assay. ** P <0.01 compared with LO2. ( F ) Expression of TMPO-AS1 in control tissues (n=50) and liver hepatocellular carcinoma (LIHC) tissues (n=374). TMPO-AS1 expression is significantly upregulated in LIHC tissues compared with control tissues based on the analysis of the high-throughput sequencing database of TCGA.

Article Snippet: HCC cells (HepG2, SNU-387, HCCLM3, SMMC-7721, Huh7) and normal human hepatic cell line, LO2 were purchased from Jiangsu KeyGEN BioTECH (Nanjing, Jiangsu, China) and were maintained in RPMI-1640 or DMEM containing 10% FBS at 37°C in a 5% CO 2 incubator.

Techniques: Expressing, Quantitative RT-PCR, Control, Next-Generation Sequencing

Journal: OncoTargets and therapy

Article Title: lncRNA TMPO-AS1 Exerts Oncogenic Roles in HCC Through Regulating miR-320a/SERBP1 Axis

doi: 10.2147/OTT.S250355

Figure Lengend Snippet: TMPO-AS1 serves as a sponge of miR-320a. ( A ) The localization of TMPO-AS1 in cells (SNU-387 and HCCLM3) was confirmed by nuclear-cytoplasmic fractionation. ( B ) The potential binding sites between miR-320a and TMPO-AS1 were hypothesized using bioinformatics analysis starBase v.2.0 database. ( C ) The targeted binding effects between miR-320a and TMPO-AS1-wt or TMPO-AS1-mut in SNU-387 and HCCLM3 cells were detected using luciferase reporter assay. ** P <0.01 compared with miR-NC. ( D ) RIP assay was conducted to confirm the binding ability between TMPO-AS1 and miR-320a. Both TMPO-AS1 and miR-320a were significantly enriched in Ago2 immunoprecipitate. ** P <0.01 compared with anti-IgG. ( E ) RNA-FISH analysis showed that miR-320a co-localizes with TMPO-AS1 in SNU-387 and HCCLM3 cells. ( F ) The relative expression of miR-320a in SNU-387 and HCCLM3 cell transfected with sh-TMPO-AS1 or pc-TMPO-AS1 was measured by RT-qPCR. ** P <0.01 compared with sh-NC. ( G ) The levels of miR-320a in HCC cells and LO2 cells were determined by qRT-PCR assay. ** P <0.01 compared with LO2. ( H ) Cell viability of SNU-387 and HCCLM3 cell after miR-320a upregulation was conducted by performing CCK-8 assay. ( I ) The proliferation of miR-320a overexpressed SNU-387 and HCCLM3 cells was examined by colony formation assay. ( J ) The migration of miR-320a overexpressed SNU-387 and HCCLM3 cells was examined by wound healing assay. ( K ) The invasion of miR-320a overexpressed SNU-387 and HCCLM3 cell was examined by Transwell invasion assay. ** P <0.01 compared with control.

Article Snippet: HCC cells (HepG2, SNU-387, HCCLM3, SMMC-7721, Huh7) and normal human hepatic cell line, LO2 were purchased from Jiangsu KeyGEN BioTECH (Nanjing, Jiangsu, China) and were maintained in RPMI-1640 or DMEM containing 10% FBS at 37°C in a 5% CO 2 incubator.

Techniques: Fractionation, Binding Assay, Luciferase, Reporter Assay, Expressing, Transfection, Quantitative RT-PCR, CCK-8 Assay, Colony Assay, Migration, Wound Healing Assay, Transwell Invasion Assay, Control

Journal: World Journal of Gastroenterology

Article Title: Effect of adipose-derived mesenchymal stem cells on hepatocellular carcinoma: In vitro inhibition of carcinogenesis

doi: 10.3748/wjg.v25.i5.567

Figure Lengend Snippet: Effect of adipose-derived mesenchymal stem cells and their conditioned media on hepatocellular carcinoma cell proliferation and apoptosis. HCC cells (2 × 10 5 ) were seeded in six-well coculture plates in the presence or absence of ADMSCs and ADMSC CM, undiluted or diluted 1:5 or 1:25 for 48 h. The proliferation of HepG2 and PLC-PRF-5 HCC cell lines were measured by (A) cell count assay and (B) WST-8 proliferation tests; The apoptosis of HepG2 (C) and PLC-PRF (D) cells co-cultured as above was measured by flow cytometry using Annexin V/PI test kit; C and D: Two representative experiments of apoptosis in HepG2 and PLC-PRF cells, respectively; E: The average rate of apoptosis in HepG2 and PLC-PRF-5 cells induced by ADMSCs, ADMSC CM. Data are representative of three independent experiments, each repeated in triplicate. All data are represented as mean ± SD ( a P < 0.05, b P < 0.01, c P < 0.001). CTR: Control; ADMSC: Adipose-derived mesenchymal stem cell; CM: Conditioned media; HCC: Hepatocellular carcinoma.

Article Snippet: The HepG2/C3A and PLC/PRF/5 cell migration assay was performed using a Boyden chamber in a 24-well plate designed by Cell Biolabs Inc. (San Diego, CA, United States) according to the manufacturer's recommendations.

Techniques: Derivative Assay, Cell Counting, Cell Culture, Flow Cytometry

Journal: World Journal of Gastroenterology

Article Title: Effect of adipose-derived mesenchymal stem cells on hepatocellular carcinoma: In vitro inhibition of carcinogenesis

doi: 10.3748/wjg.v25.i5.567

Figure Lengend Snippet: Effect of adipose derived mesenchymal stem cells and their conditioned media on of hepatocellular carcinoma cell migration and invasion. HCC cells (2 × 10 5 ) were seeded in six- well co-culture plates in the presence or absence of ADMSCs (at ADSMCs: HCC ratio of 1:1 or 1:2) or ADMSC CM, undiluted or diluted 1:5 or 1:25. The migration of (A) HepG2 and (B) PLC-PRF-5 cells was assessed by wound healing assay. The migration rate at 24 h is represented in (C); D: A Transwell migration assay was performed to confirm the results of the wound healing assay; E: HCC cell invasiveness was measured by Transwell invasion assay. In the Transwell migration and invasion assay, 3 × 10 5 HCC cells alone, co-cultured with ADMSCs, or treated with ADMSC CM were seeded into the apical chamber of Transwell plates and allowed to migrate or invade through the uncoated polycarbonate membrane or collagen-coated polycarbonate membrane, respectively (8 μm pore size) to the lower chamber for 24 h or 48 h, respectively. The migratory or invasive cells were stained with crystal violet cell stain solution and extracted using an extraction solution provided in the kit. The level of migration and invasion was measured using a plate reader at the absorbance of 560 nm. Values shown are representative of five independent experiments, each performed in triplicate. Data are represented as mean ± SD of five independent experiments, each performed in triplicate ( a P < 0.05, b P < 0.01, c P < 0.001). CTR: Control; ADMSC: Adipose derived mesenchymal stem cell; CM: Conditioned media; HCC: Hepatocellular carcinoma.

Article Snippet: The HepG2/C3A and PLC/PRF/5 cell migration assay was performed using a Boyden chamber in a 24-well plate designed by Cell Biolabs Inc. (San Diego, CA, United States) according to the manufacturer's recommendations.

Techniques: Derivative Assay, Migration, Co-Culture Assay, Wound Healing Assay, Transwell Migration Assay, Transwell Invasion Assay, Invasion Assay, Cell Culture, Staining

Journal: World Journal of Gastroenterology

Article Title: Effect of adipose-derived mesenchymal stem cells on hepatocellular carcinoma: In vitro inhibition of carcinogenesis

doi: 10.3748/wjg.v25.i5.567

Figure Lengend Snippet: Effect of adipose-derived mesenchymal stem cells and adipose-derived mesenchymal stem cell conditioned media on tissue inhibitor metalloproteinase mRNA levels in hepatocellular carcinoma cells. HCC cells (2 × 10 5 ) were seeded in six-well co-culture plates in the presence or absence of ADMSCs or of undiluted ADMSC conditioned media (CM). The mRNA levels of TIMP-1, TIMP-2, and TIMP-3 in HepG2 (A) and PLC-PRF-5 (B) cells after the removal of ADMSCs in the case of coculture was measured by quantitative PCR. Data are represented as mean ± SD of five independent experiments, each performed in triplicate ( a P < 0.05, b P < 0.01, c P < 0.001). ADMSC: Adipose-derived mesenchymal stem cell; CM: Conditioned media; HCC: Hepatocellular carcinoma.

Article Snippet: The HepG2/C3A and PLC/PRF/5 cell migration assay was performed using a Boyden chamber in a 24-well plate designed by Cell Biolabs Inc. (San Diego, CA, United States) according to the manufacturer's recommendations.

Techniques: Derivative Assay, Co-Culture Assay, Real-time Polymerase Chain Reaction

Journal: World Journal of Gastroenterology

Article Title: Effect of adipose-derived mesenchymal stem cells on hepatocellular carcinoma: In vitro inhibition of carcinogenesis

doi: 10.3748/wjg.v25.i5.567

Figure Lengend Snippet: Expression of tumor suppressor genes and oncogenes in hepatocellular carcinoma cells. HCC cells (2 × 10 5 ) were seeded in six-well co-culture plates in the presence or absence of ADMSCs or undiluted ADMSC CM for 48 h. The mRNA expression of the tumor suppressor genes P53/RB, oncogene c-Myc and the enzymatic component of telomerase hTERT were assessed by RT-PCR in (A) HepG2 and (B) PLC-PRF-5 cells after the removal of ADMSCs in the case of co-culture. Data are represented as mean ± SD of five independent experiments, each performed in triplicate ( a P < 0.05, b P < 0.01, c P < 0.001). ADMSC: Adipose-derived mesenchymal stem cell; CM: Conditioned media; HCC: Hepatocellular carcinoma.

Article Snippet: The HepG2/C3A and PLC/PRF/5 cell migration assay was performed using a Boyden chamber in a 24-well plate designed by Cell Biolabs Inc. (San Diego, CA, United States) according to the manufacturer's recommendations.

Techniques: Expressing, Co-Culture Assay, Reverse Transcription Polymerase Chain Reaction, Derivative Assay