Journal: Oncology Letters

Article Title: HOTAIR contributes to the growth of liver cancer via targeting miR-217

doi: 10.3892/ol.2018.8341

Figure Lengend Snippet: Expression of HOTAIR was upregulated in liver cancer tissues and liver cancer cell lines. (A) The expression of HOTAIR in tumor tissues and normal tissues was detected by qRT-RCR. (B) The expression of HOTAIR in 3 liver cancer cell lines (MHCC 97H, HepG2 and Hep3B) and normal human hepatic cell line HL-7702 was evaluated by qRT-RCR. (C) The expression of HOTAIR in tumor tissues and normal tissues was measured by northern blotting. (D) The expression of HOTAIR in MHCC 97H, HepG2, Hep3B and HL-7702 cell lines was measured by northern blotting (*P<0.05, **P<0.01). HOTAIR, HOX transcript antisense RNA

Article Snippet: The normal human hepatic cell line HL-7702 and liver cancer cell lines (MHCC 97H, HepG2 and Hep3B) were obtained from American Type Culture Collection (Manassas, VA, USA).

Techniques: Expressing, Northern Blot

Journal: Oncology Letters

Article Title: HOTAIR contributes to the growth of liver cancer via targeting miR-217

doi: 10.3892/ol.2018.8341

Figure Lengend Snippet: HOTAIR inhibition suppressed the proliferation of HepG2 cells. HepG2 cells were infected with HOTAIR siRNA lentiviral vectors or the empty lentiviral vectors for 72 h. (A) The expression of HOTAIR was evaluated by qRT-RCR. (B) The proliferation rate was measured by CCK-8 assay. (C and D) The cell number was assessed by optical microscope. (E) The protein expression of two proliferation marker proteins Ki67 and PCNA was evaluated by western blotting. GAPDH was used as an internal control (*P<0.05, ***P<0.001). HOTAIR, HOX transcript antisense RNA.

Article Snippet: The normal human hepatic cell line HL-7702 and liver cancer cell lines (MHCC 97H, HepG2 and Hep3B) were obtained from American Type Culture Collection (Manassas, VA, USA).

Techniques: Inhibition, Infection, Expressing, CCK-8 Assay, Microscopy, Marker, Western Blot

Journal: Oncology Letters

Article Title: HOTAIR contributes to the growth of liver cancer via targeting miR-217

doi: 10.3892/ol.2018.8341

Figure Lengend Snippet: HOTAIR directly targeted miR-217. (A) HepG2 cells were transfected with miR-217 mimics or miR-217 mock alone, or co-transfected with HOTAIR siRNA lentiviral vectors together. The expression of miR-217 was evaluated by northern blotting. (B) HepG2 cells were transfected with miR-217 inhibitors or miR-217 mock alone, or co-transfected with HOTAIR siRNA lentiviral vectors together. The expression of miR-217 was evaluated by northern blotting. (C) HepG2 cells were co-transfected with pGL3 plasmids containing HOTAIR (WT or MUT) sequences and miR-217 mimics. The luciferase activity was detected by luciferase reporter assay. (D) The expression correlation between HOTAIR and mir-217 was determined in 25 liver cancer tissues (**P<0.01). HOTAIR, HOX transcript antisense RNA. miR-217, microRNA-217.

Article Snippet: The normal human hepatic cell line HL-7702 and liver cancer cell lines (MHCC 97H, HepG2 and Hep3B) were obtained from American Type Culture Collection (Manassas, VA, USA).

Techniques: Transfection, Expressing, Northern Blot, Luciferase, Activity Assay, Reporter Assay

Journal: Oncology Letters

Article Title: HOTAIR contributes to the growth of liver cancer via targeting miR-217

doi: 10.3892/ol.2018.8341

Figure Lengend Snippet: miR-217 was involved in the regulation of HOTAIR on cell proliferation and cycle arrest in HepG2 cells. HepG2 cells were transfected with HOTAIR siRNA lentiviral vectors or miR-217 inhibitors, or the combination. (A) The proliferation rate was measured by CCK-8 assay. (B) The cell number was assessed by optical microscope. (C) The protein expression of Ki67, PCNA, p27 and cyclin D1 was evaluated by western blotting. (D) The cell cycle arrest was evaluated by flow cytometry (**P<0.01). HOTAIR, HOX transcript antisense RNA. miR-217, microRNA-217.

Article Snippet: The normal human hepatic cell line HL-7702 and liver cancer cell lines (MHCC 97H, HepG2 and Hep3B) were obtained from American Type Culture Collection (Manassas, VA, USA).

Techniques: Transfection, CCK-8 Assay, Microscopy, Expressing, Western Blot, Flow Cytometry

Journal: Oncology Letters

Article Title: HOTAIR contributes to the growth of liver cancer via targeting miR-217

doi: 10.3892/ol.2018.8341

Figure Lengend Snippet: HOTAIR inhibition suppresses tumor growth in xenograft models. Xenograft mouse model was created by subcutaneous injection of HepG2 cells pretreated with or without HOTAIR siRNA lentiviral vectors to nude mouse. (A) Five primary tumors from each group were photographed after sacrifice at 30th day. (B) Tumor growth curves in two groups were shown every 6 days until the 30th day following cells injection. (C and D) The expression of HPTAIR and miR-217 in primary tumor tissues was measured by qRT-PCR. (E and F) The expression of HPTAIR and miR-217 in primary tumor tissues was measured by northern blotting. (G and H) The expression of Ki67 and PCNA in primary tumor tissues was visualized by immunohistochemistry (*P<0.05, **P<0.01). HOTAIR, HOX transcript antisense RNA. miR-217, microRNA-217.

Article Snippet: The normal human hepatic cell line HL-7702 and liver cancer cell lines (MHCC 97H, HepG2 and Hep3B) were obtained from American Type Culture Collection (Manassas, VA, USA).

Techniques: Inhibition, Injection, Expressing, Quantitative RT-PCR, Northern Blot, Immunohistochemistry

Journal: International Journal of Molecular Sciences

Article Title: Lacticaseibacillus paracsei HY7207 Alleviates Hepatic Steatosis, Inflammation, and Liver Fibrosis in Mice with Non-Alcoholic Fatty Liver Disease

doi: 10.3390/ijms25189870

Figure Lengend Snippet: Lists of TaqMan probes used for gene expression analysis.

Article Snippet: The expression levels of the target genes were normalized to that of GAPDH (HepG2 cells, Hs99999905_m1; Mice, Mm99999915_g1) using the comparative C τ method.

Techniques: Gene Expression, In Vitro, In Vivo

Journal: Oncology Letters

Article Title: Exosomes derived from endoplasmic reticulum-stressed liver cancer cells enhance the expression of cytokines in macrophages via the STAT3 signaling pathway

doi: 10.3892/ol.2020.11609

Figure Lengend Snippet: Characteristics of ER stress-associated exosomes. (A) HepG2 cells were treated with 0, 1.25, 2.5 and 5 µM TM for 24 h, and (B) GRP78 protein expression was measured and semi-quantitatively analyzed, relative to β-actin intensity. (C) HepG2 cells were treated with 2.5 µM TM for 12, 24 and 48 h, and (D) GRP78 protein expression was measured and semi-quantitatively analyzed, relative to β-actin intensity. (E) Representative transmission electron microscope images of Exo-TM. Scale bar, 100 nm. (F) Expression levels of CD63, TSG101, CD81, β-actin and Calnexin were measured using cell lysates or exosomes by western blotting. *P< 0.05, **P< 0.01. ER, endoplasmic reticulum; TM, tuniamycin; GRP78, glucose-regulated protein 78; Exo-TM, exosomes from the supernatants of HepG2 cells treated with 2.5 µM tunicamycin; TSG101, tumor susceptibility gene 101; Exo-con, exosomes from the supernatants of control HepG2 cells.

Article Snippet: Exosomes from the supernatants of normal cultured HepG2 cells (Exo-con) and HepG2 cells treated with 2.5 μM tunicamycin (Sigma-Aldrich; Merck KGaA) (Exo-TM) at 37°C for 24 h were purified using ExoQuick Precipitation Solution (System Biosciences, LLC) at the volume ratio of 1:5, according to the manufacturer's protocol.

Techniques: Expressing, Transmission Assay, Microscopy, Western Blot, Control

Journal: Oncology Letters

Article Title: Exosomes derived from endoplasmic reticulum-stressed liver cancer cells enhance the expression of cytokines in macrophages via the STAT3 signaling pathway

doi: 10.3892/ol.2020.11609

Figure Lengend Snippet: Incubation with Exo-TM increases expression of cytokines in macrophages in vitro . (A) Confocal microscopy was used to measure the incorporation of PKH67-labeled exosomes into RAW264.7 cells. Cells were incubated with Exo-con and Exo-TM for 24 h, and (B) IL-6, (C) IL-10, (D) MCP-1 and (E) TNF-α levels were measured using a CBA inflammatory factor kit. *P< 0.05, **P< 0.01 with indicated groups. Exo-Con, exosomes from the supernatants of control HepG2 cells; Exo-TM, exosomes from the supernatants of HepG2 cells treated with 2.5 µM tunicamycin; IL, interleukin; MCP-1, monocyte chemotactic protein-1; TNF-α, tumor necrosis factor-α; CBA, cytokine bead array.

Article Snippet: Exosomes from the supernatants of normal cultured HepG2 cells (Exo-con) and HepG2 cells treated with 2.5 μM tunicamycin (Sigma-Aldrich; Merck KGaA) (Exo-TM) at 37°C for 24 h were purified using ExoQuick Precipitation Solution (System Biosciences, LLC) at the volume ratio of 1:5, according to the manufacturer's protocol.

Techniques: Incubation, Expressing, In Vitro, Confocal Microscopy, Labeling, Control

Journal: Oncology Letters

Article Title: Exosomes derived from endoplasmic reticulum-stressed liver cancer cells enhance the expression of cytokines in macrophages via the STAT3 signaling pathway

doi: 10.3892/ol.2020.11609

Figure Lengend Snippet: Exo-TM enhances cytokine expression by activating the JAK2/STAT3 pathway. RAW264.7 cells were treated with Exo-con or Exo-TM for 24 h. (A) Protein expression levels of the JAK2/STAT3 pathway were measured by western blot analysis. Blots presented in (A) were semi-quantitatively analyzed for comparing the (B) p-STAT3/STAT3 and (C) p-JAK2/JAK2 ratios, relative to β-actin intensity. (D) Western blot analysis of the protein expression levels of members of the JAK2/STAT3 signaling pathway in RAW264.7 cells treated with Exo-TM and S3I-201. Blots presented in (D) were semi-quantitatively analyzed for comparing the (E) p-STAT3/STAT3 and (F) p-JAK2/JAK2 ratios, relative to β-actin intensity. *P< 0.05, **P< 0.01 with indicated groups. Exo-Con, exosomes from the supernatants of control HepG2 cells; Exo-TM, exosomes from the supernatants of HepG2 cells treated with 2.5 µM tunicamycin; TM, tunicamycin; JAK2, Janus kinase 2; p-, phosphorylated.

Article Snippet: Exosomes from the supernatants of normal cultured HepG2 cells (Exo-con) and HepG2 cells treated with 2.5 μM tunicamycin (Sigma-Aldrich; Merck KGaA) (Exo-TM) at 37°C for 24 h were purified using ExoQuick Precipitation Solution (System Biosciences, LLC) at the volume ratio of 1:5, according to the manufacturer's protocol.

Techniques: Expressing, Western Blot, Control

Journal: Oncology Letters

Article Title: Exosomes derived from endoplasmic reticulum-stressed liver cancer cells enhance the expression of cytokines in macrophages via the STAT3 signaling pathway

doi: 10.3892/ol.2020.11609

Figure Lengend Snippet: Inhibition of STAT3 decreases Exo-TM-induced cytokine expression. RAW264.7 macrophages were treated with Exo-con or Exo-TM with or without S3I-201 for 24 h. (A) IL-6, (B) IL-10, (C) MCP-1 and (D) TNF-α levels were measured using a CBA inflammatory factor kit. *P< 0.05, **P< 0.01 with indicated groups. Exo-Con, exosomes from the supernatants of control HepG2 cells; Exo-TM, exosomes from the supernatants of HepG2 cells treated with 2.5 µM tunicamycin; IL, interleukin; MCP-1, monocyte chemotactic protein-1; TNF-α, tumor necrosis factor-α; CBA, cytokine bead array.

Article Snippet: Exosomes from the supernatants of normal cultured HepG2 cells (Exo-con) and HepG2 cells treated with 2.5 μM tunicamycin (Sigma-Aldrich; Merck KGaA) (Exo-TM) at 37°C for 24 h were purified using ExoQuick Precipitation Solution (System Biosciences, LLC) at the volume ratio of 1:5, according to the manufacturer's protocol.

Techniques: Inhibition, Expressing, Control