Journal: Scientific Reports

Article Title: α-Lipoic acid induces Endoplasmic Reticulum stress-mediated apoptosis in hepatoma cells

doi: 10.1038/s41598-020-64004-5

Figure Lengend Snippet: α-LA induces ER stress in hepatoma cells. ( A ) Western analysis of key players in UPR after treatment of HepG2 with 500 µM α-LA from 6 up to 48 hours. Albumin was used as loading control. For densitometric analysis protein expression was normalized to Albumin expression and values (reported over each band) have been expressed as fold change respect to control. Each lane represents a pool of three individual samples. ( B ) Schematic representation of α-LA-mediated apoptosis in hepatoma cells. Western blot images ( A ) have been cropped for clarity with full blot presented in Supplementary Fig. .

Article Snippet: The rat hepatoma cell line, FaO, and the hepatocarcinoma cell line, HepG2, were supplied by Interlab Cell Line Collection (Servizio Biotecnologie, IST, Genova, Italy), and maintained, respectively, in Dulbecco’s medium (DMEM plus Glutamax I) (Invitrogen) and supplemented with penicillin, streptomycin and 10% heat-inactivated fetal calf-serum (FCS) (Invitrogen) in a humidified atmosphere of 5% CO 2 /95% air, at 37 °C. α-Lipoic Acid (α-LA) and Thapsigargin (TG) were purchased from Sigma (Sigma-Aldrich, Milano, Italy). α-LA, dissolved in sodium hydroxide NaOH 1 N and neutralized in medium, and TG dissolved in DMSO, were added to the culture media to the final concentrations specified in the text.

Techniques: Western Blot, Control, Expressing

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

Journal: PLoS ONE

Article Title: Cellular Trafficking of Thymosin Beta-4 in HEPG2 Cells Following Serum Starvation

doi: 10.1371/journal.pone.0067999

Figure Lengend Snippet: Tβ 4 immunoreactivity in HepG2 cells cultured for 48 h with serum (A and B, magnification 1000×) and without serum (C and D, magnification 1000×). Inserts in A and in C represent one particular of the intranuclear reactivity of the respective pictures (magnification 1000×).

Article Snippet: Commercial human cell line HepG2 (ICLC HTL95005), were obtained from the Istituto Nazionale per la Ricerca sul Cancro c/o CBA (ICLC, Genova).

Techniques: Cell Culture

Journal: PLoS ONE

Article Title: Cellular Trafficking of Thymosin Beta-4 in HEPG2 Cells Following Serum Starvation

doi: 10.1371/journal.pone.0067999

Figure Lengend Snippet: Tβ 4 immunoreactivity in HepG2 cells cultured for 72 h with serum (A and B, magnification 1000×) and without serum (C and D, magnification respectively 400 and 1000×). Inserts in A and in C represent one particular of the intranuclear reactivity of the respective pictures (magnification 1000×).

Article Snippet: Commercial human cell line HepG2 (ICLC HTL95005), were obtained from the Istituto Nazionale per la Ricerca sul Cancro c/o CBA (ICLC, Genova).

Techniques: Cell Culture

Journal: PLoS ONE

Article Title: Cellular Trafficking of Thymosin Beta-4 in HEPG2 Cells Following Serum Starvation

doi: 10.1371/journal.pone.0067999

Figure Lengend Snippet: Tβ 4 immunoreactivity in HepG2 cells cultured for 84 h with serum (A and B, magnification 1000×) and without serum (C and D, magnification 1000×). Inserts in A and in C represent one particular of the intranuclear reactivity of the respective pictures (magnification 1000×).

Article Snippet: Commercial human cell line HepG2 (ICLC HTL95005), were obtained from the Istituto Nazionale per la Ricerca sul Cancro c/o CBA (ICLC, Genova).

Techniques: Cell Culture

Journal: PLoS ONE

Article Title: Cellular Trafficking of Thymosin Beta-4 in HEPG2 Cells Following Serum Starvation

doi: 10.1371/journal.pone.0067999

Figure Lengend Snippet: Tβ 4 immunoreactivity in HepG2 cells cultured for 84 h without serum and shifted in complete medium for 24 h (A and B, magnification 400×) and for 48 h (C and D, magnification 400×). Inserts in A and in C represent one particular of the intranuclear reactivity of the respective pictures (magnification 1000×).

Article Snippet: Commercial human cell line HepG2 (ICLC HTL95005), were obtained from the Istituto Nazionale per la Ricerca sul Cancro c/o CBA (ICLC, Genova).

Techniques: Cell Culture