Journal: Toxics

Article Title: Machine Learning Model for Prediction of Development of Cancer Stem Cell Subpopulation in Tumurs Subjected to Polystyrene Nanoparticles

doi: 10.3390/toxics12050354

Figure Lengend Snippet: Summary of in vitro and in vivo studies using polystyrene particles.

Article Snippet: In vivo : BALB/c nude mice In vitro : human gastric cancer cell lines (AGS, MKN1, MKN45, NCI-N87, and KATOIII) [ ] , purchased from Cospheric (Somis, CA, USA) , polystyrene , 9.5–11.5 μm , In vivo : 1.72 × 10 4 particles/mL In vitro: 8.61 × 10 5 particles/mL , induced resistance to chemo- and monoclonal antibody-therapy.

Techniques: In Vitro, In Vivo, Polymer, Concentration Assay, Mouse Assay

Journal: Biomedicines

Article Title: Isoliquiritigenin Inhibits Gastric Cancer Stemness, Modulates Tumor Microenvironment, and Suppresses Tumor Growth through Glucose-Regulated Protein 78 Downregulation

doi: 10.3390/biomedicines10061350

Figure Lengend Snippet: Glucose-regulated protein 78 (GRP78) was overexpressed in spheroid gastric cancer cells. ( A ) A representative picture demonstrating the MKN45 cells with the spheroid-forming ability. ( B ) Protein expression of GRP78 in parental and spheroid body-forming MKN45 cells. ( C ) Quantification of GRP78 expression in parental and spheroid body–forming MKN45 cells. Data are expressed as the mean ± standard error of mean; n ≥ 3 independent experiments, two-tailed Student’s t test: ** p < 0.01.

Article Snippet: The MKN45 cells stably transfected with GRP78 shRNA or scrambled shRNA were harvested and subcutaneously inoculated (5 × 10 6 cells/0.1 mL in PBS) into 6-week-old BALB/c nude mice (BioLASCO, Taipei City, Taiwan).

Techniques: Expressing, Two Tailed Test

Journal: Biomedicines

Article Title: Isoliquiritigenin Inhibits Gastric Cancer Stemness, Modulates Tumor Microenvironment, and Suppresses Tumor Growth through Glucose-Regulated Protein 78 Downregulation

doi: 10.3390/biomedicines10061350

Figure Lengend Snippet: Glucose-regulated protein 78 (GRP78) promoted stemness in MKN45 cells. ( A ) Representative pictures showing spheroid body-forming and ( B ) colony-forming MKN45/ctrl and MKN45/GRP78 + cells. ( C ) Stemness-related surface makers (LGR5, CD24, and CD44) were analyzed through flow cytometry in MKN45/ctrl and MKN45/GRP78 + cells. ( D ) Quantification of the expression of surface makers (LGR5, CD24, and CD44). ( E ) Aldehyde dehydrogenase 1-positive cells were analyzed through flow cytometry among MKN45/ctrl and MKN45/GRP78 + cells. ( F ) Protein expression of stemness-related transcription factors (SOX2 and Nanog) and GRP78 in MKN45/ctrl and MKN45/GRP78 + cells. ( G ) Quantification of SOX2, Nanog, and GRP78 expression in MKN45/ctrl and MKN45/GRP78 + cells. Data are expressed as the mean ± standard error of mean; n ≥ 3 independent experiments, two-tailed Student’s t test: * p < 0.05, ** p < 0.01.

Article Snippet: The MKN45 cells stably transfected with GRP78 shRNA or scrambled shRNA were harvested and subcutaneously inoculated (5 × 10 6 cells/0.1 mL in PBS) into 6-week-old BALB/c nude mice (BioLASCO, Taipei City, Taiwan).

Techniques: Flow Cytometry, Expressing, Two Tailed Test

Journal: Biomedicines

Article Title: Isoliquiritigenin Inhibits Gastric Cancer Stemness, Modulates Tumor Microenvironment, and Suppresses Tumor Growth through Glucose-Regulated Protein 78 Downregulation

doi: 10.3390/biomedicines10061350

Figure Lengend Snippet: The stemness characteristic was reduced by isoliquiritigenin (ISL) treatment. ( A ) Structural formula of ISL. ( B ) Glucose-regulated protein 78 (GRP78) expression in MKN45 cells was inhibited after treatment with 15 or 25 μg/mL ISL for 72 h. ( C ) Quantification of GRP78 expression after ISL treatment. ( D ) The protein expression of the transcription factor CREB3L1 was downregulated in MKN45 cells after treatment with 15 or 25 μg/mL ISL for 72 h. ( E ) Quantification of CREB3L1. ( F ) Representative pictures demonstrating that the colony-forming MKN45 cells were decreased after treatment with 25 μg/mL ISL. ( G ) Representative pictures demonstrating that the spheroid body-forming MKN45 cells were decreased after treatment with 25 μg/mL ISL. ( H , I ) The protein expression of stemness-related transcription factors (SOX2 and Nanog) was downregulated in MKN45 cells after treatment with 15 or 25 μg/mL ISL for 72 h. Data are presented as the mean ± standard error of mean; n ≥ 3 independent experiments, two-tailed Student’s t test: * p < 0.05, ** p < 0.01, *** p < 0.005.

Article Snippet: The MKN45 cells stably transfected with GRP78 shRNA or scrambled shRNA were harvested and subcutaneously inoculated (5 × 10 6 cells/0.1 mL in PBS) into 6-week-old BALB/c nude mice (BioLASCO, Taipei City, Taiwan).

Techniques: Expressing, Two Tailed Test

Journal: Biomedicines

Article Title: Isoliquiritigenin Inhibits Gastric Cancer Stemness, Modulates Tumor Microenvironment, and Suppresses Tumor Growth through Glucose-Regulated Protein 78 Downregulation

doi: 10.3390/biomedicines10061350

Figure Lengend Snippet: Glucose-regulated protein 78 (GRP78) silencing reduced the stemness capacity of MKN45 cells. ( A ) Representative pictures showing the spheroid-forming capacity between MKN45/ctrl and MKN45/sh-GRP78 cells. ( B ) Representative pictures showing the colony-forming capacity between MKN45/ctrl and MKN45/sh-GRP78 cells. ( C ) Expression of surface makers (LGR5, CD24, and CD44) was analyzed through flow cytometry in MKN45/ctrl and MKN45/sh-GRP78 cells. ( D ) Quantification of surface makers. ( E ) Aldehyde dehydrogenase 1-positive cells were analyzed through flow cytometry in MKN45/ctrl and MKN45/sh-GRP78 cells. ( F ) Protein expression of stemness-related transcription factors (SOX2 and Nanog) and GRP78 was analyzed through Western blot in MKN45/ctrl and MKN45/sh-GRP78 cells. ( G ) Quantification of the expression of stemness markers (SOX2 and Nano) and GRP78. Data are presented as the mean ± standard error of the mean; n ≥ 3 independent experiments, two-tailed Student’s t test: * p < 0.05, ** p < 0.01, *** p < 0.005.

Article Snippet: The MKN45 cells stably transfected with GRP78 shRNA or scrambled shRNA were harvested and subcutaneously inoculated (5 × 10 6 cells/0.1 mL in PBS) into 6-week-old BALB/c nude mice (BioLASCO, Taipei City, Taiwan).

Techniques: Expressing, Flow Cytometry, Western Blot, Two Tailed Test

Journal: Biomedicines

Article Title: Isoliquiritigenin Inhibits Gastric Cancer Stemness, Modulates Tumor Microenvironment, and Suppresses Tumor Growth through Glucose-Regulated Protein 78 Downregulation

doi: 10.3390/biomedicines10061350

Figure Lengend Snippet: Knockdown of glucose-regulated protein 78 (GRP78) reduced tumor growth in gastric cancer xenografts. ( A ) Schematic of gastric cancer xenografts between MKN45/ctrl and MKN45/shGRP78 groups. ( B ) The curves of tumor growth in mice between MKN45/ctrl and MKN45/shGRP78 groups. ( C ) Representative images of hematoxylin and eosin and immunohistochemical (IHC) staining of ki-67 between MKN45/ctrl and MKN45/shGRP78 groups. ( D ) Representative IHC analysis of GRP78 in MKN45/ctrl and MKN45/shGRP78 gastric cancer xenografts and relative quantification per intensity of staining scoring. ( E ) Representative IHC analysis of ki-67 staining in MKN45/ctrl and MKN45/shGRP78 gastric cancer xenografts and relative quantification per intensity of staining scoring. ( F ) Curves of body weight of mice between MKN45/ctrl and MKN45/shGRP78 groups. ( G ) Tumor sizes of each group. Data are presented as the mean ± standard error of mean; n ≥ 4 independent experiments, two-tailed Student’s t test: ** p < 0.01, *** p < 0.005.

Article Snippet: The MKN45 cells stably transfected with GRP78 shRNA or scrambled shRNA were harvested and subcutaneously inoculated (5 × 10 6 cells/0.1 mL in PBS) into 6-week-old BALB/c nude mice (BioLASCO, Taipei City, Taiwan).

Techniques: Immunohistochemical staining, Immunohistochemistry, Staining, Two Tailed Test

Journal: Biomedicines

Article Title: Isoliquiritigenin Inhibits Gastric Cancer Stemness, Modulates Tumor Microenvironment, and Suppresses Tumor Growth through Glucose-Regulated Protein 78 Downregulation

doi: 10.3390/biomedicines10061350

Figure Lengend Snippet: Glucose-regulated protein 78 (GRP78) expression in gastric cancer cells induces cancer-associated fibroblast activity. ( A ) Levels of tumor growth factor (TGF)-β1 in the condition medium of MKN45/ctrl, MKN45/GRP78 + , and MKN45/sh-GRP78 cells. ( B ) Levels of TGF-β1 in the condition medium in MKN45 cells with or without ISL treatment. ( C ) Protein expression of α-SMA and matrix metalloproteinase (MMP-9) was analyzed through a Western blot after treatment with 15 and 25 μg/mL isoliquiritigenin (ISL) for 48 h in h-GCA N3 cells. ( D , E ) Quantification of α-SMA and MMP-9 expression was analyzed through a Western blot after treatment with 15 and 25 μg/mL ISL for 48 h. ( F ) Levels of IL-6 in the conditioned medium in h-GCA N3 cells with or without ISL treatment. ( G ) Levels of TGF-β1 in the conditioned medium in h-GCA N3 cells with or without ISL treatment. Data are presented as the mean ± standard error of mean; n ≥ 3 independent experiments, two-tailed Student’s t test: * p < 0.05, ** p < 0.01, *** p < 0.005.

Article Snippet: The MKN45 cells stably transfected with GRP78 shRNA or scrambled shRNA were harvested and subcutaneously inoculated (5 × 10 6 cells/0.1 mL in PBS) into 6-week-old BALB/c nude mice (BioLASCO, Taipei City, Taiwan).

Techniques: Expressing, Activity Assay, Western Blot, Two Tailed Test

Journal: Biomedicines

Article Title: Isoliquiritigenin Inhibits Gastric Cancer Stemness, Modulates Tumor Microenvironment, and Suppresses Tumor Growth through Glucose-Regulated Protein 78 Downregulation

doi: 10.3390/biomedicines10061350

Figure Lengend Snippet: Isoliquiritigenin (ISL) inhibited 5-fluorouracil (5-FU)-induced glucose-regulated protein 78 (GRP78)-mediated stemness. ( A ) GRP78 expression was analyzed through Western blot after treatment with 0.5 μM 5-FU for 72 h. ( B ) Quantification of GRP78 expression. ( C ) Expression of surface markers (LGR5, CD24, and CD44) was analyzed through flow cytometry in MKN45 cells after treatment with ISL or ISL combined with 5-FU. ( D ) Quantification of surface makers. ( E , F ) Representative pictures showing the spheroid-forming capacity of MKN45 cells treated with or without ISL combined with 5-FU. ( G ) The viability of the MKN45, ISL-treated MKN45, MKN45/sh-GRP78, 5-FU-treated MKN45, 5-FU+ISL-treated MKN45, and 5-Fu-treated MKN45/sh-GRP78 cells was evaluated. Data are presented as the mean ± standard error of mean; n ≥ 3 independent experiments, two-tailed Student’s t test: * p < 0.05, ** p < 0.01, *** p < 0.005.

Article Snippet: The MKN45 cells stably transfected with GRP78 shRNA or scrambled shRNA were harvested and subcutaneously inoculated (5 × 10 6 cells/0.1 mL in PBS) into 6-week-old BALB/c nude mice (BioLASCO, Taipei City, Taiwan).

Techniques: Expressing, Western Blot, Flow Cytometry, Two Tailed Test