Journal: Cancer Management and Research

Article Title: ATG3, a Target of miR-431-5p, Promotes Proliferation and Invasion of Colon Cancer via Promoting Autophagy

doi: 10.2147/CMAR.S226828

Figure Lengend Snippet: ATG3 knockdown inhibits proliferation and invasion of colon cancer cells. Notes: qPCR ( A ) and Western blot ( B ) show that ATG3 is upregulated in colon cancer cells compared with colon epithelial cell NCM460. ( C ) Western blot indicates that ATG3 is successfully knocked down in SW620 and HCT116 cells. ATG3 knockdown significantly suppresses proliferation and invasion of colon cancer cells demonstrated by plate clone formation ( D ) and Transwell assays ( E ). siNC stands for negative control small RNA; Ns stands for no significant difference; **Stands for P < 0.01; ***Stands for P < 0.001.

Article Snippet: NCM460, an immortalized colon epithelial cell line, was purchased from ATCC and cultured with M3 Base medium (INCELL, TX, USA) plus 10% FBS (BI, Jerusalem, Israel).

Techniques: Knockdown, Western Blot, Negative Control

Journal:

Article Title: Ineffective Vaccination Against Solid Tumors Can Be Enhanced by Hematopoetic Cell Transplantation

doi: 10.4049/jimmunol.0900159

Figure Lengend Snippet: A, Experimental scheme. Syngeneic Thy 1.1 donors were vaccinated with 106 irradiated CT26 cells and CpG. Bone marrow and splenocytes were transplanted into lethally irradiated Thy 1.2 hosts with s.c. tumors established for 14 days. Tumor-infiltrating cells and host spleens were analyzed d 14 after HCT (28 days after tumors were induced). Control tumor-bearing animals did not receive irradiation and HCT. B, Analysis of CD62L expression on CD8+ and CD4+ tumor-infiltrating T cells form irradiated hosts that received HCT (gated Thy1.1+ cells) or untreated control animals (no HCT)(gated Thy1.2+cells) at day 28. The data are representative for the group of animals (n=5).

Article Snippet: Cell lines The CT26 cell line (an N-nitro-N-methylurethane-induced BALB/c murine colon carcinoma) was purchased from ATCC (Manassas, VA).

Techniques: Irradiation, Control, Expressing

Journal: Molecular cancer therapeutics

Article Title: Apigenin sensitizes colon cancer cells to anti-tumor activity of ABT-263

doi: 10.1158/1535-7163.MCT-13-0066

Figure Lengend Snippet: A, HCT116 cells were treated for 1 day with ABT-263 (ABT, 1 µM) in the presence of indicated concentrations of apigenin (apg) (left) or with 20 µM apigenin along with indicated concentrations of ABT-263 (right). Cell death was determined with the TC10™ Automatic Cell Counter as described in Materials and Methods. B, HCT116 cells were treated with ABT-263 (0.5 µM), apigenin (20 µM) alone or their combination (comb) for the indicated periods of time and cell death was quantified as in A. C, Combination index (CI) in HCT116 cells was calculated. D, The multiple colon cancer cell lines HCT116, DLD1, SW48, HT29 and HCT-8 were treated for 2 days with ABT-263 (0.5, 0.5, 0.25, 1 and 0.5 µM respectively), apigenin (20 µM), or their combination and cell death rates were determined as in A. E, Cellular apoptosis in HCT116 and DLD1 cells treated for 1 day with ABT-263 (0.5 µM), apigenin (20 µM), or their combination was determined by flow cytometry using PI and FITC-Annexin V staining. F, western blotting analysis was conducted to examine caspase 3 cleavage in HCT116 and DLD1 cells treated as in E. For this and following figures, statistical significances of data were indicted by * if p < 0.05 or by ** if p < 0.01.

Article Snippet: Lentiviral plasmid of pCDH1-HA-Mcl-1 and its control pCDH1-EF1-puro were gifts of Dr. Shengbing Huang (Mayo Clinic, MN) Cell lines The human colon cancer cell lines DLD1, HCT116, HCT-8, HT29 and SW48 were obtained from American Type Culture Collection (Manassas, VA).

Techniques: Flow Cytometry, Staining, Western Blot, IF-P

Journal: Molecular cancer therapeutics

Article Title: Apigenin sensitizes colon cancer cells to anti-tumor activity of ABT-263

doi: 10.1158/1535-7163.MCT-13-0066

Figure Lengend Snippet: A, HCT116 and DLD1cells were treated for 6 hours with the indicated doses of apigenin (upper panel) or treated with apigenin (20 µM) for indicated periods of time (lower panel). Mcl-1 protein expression was examined by immunoblotting. B, Mcl-1 protein expression was examined in HCT116 and DLD1 cells treated for 12 hours with apigenin (20 µM), ABT-263 (0.5 µM) or their combination. C, Mcl-1 expression in HCT116 cells was donwregulated by lentivirus-mediated shRNA. The Mcl-1 knockdown cells (sh-Mcl-1) and the control shRNA-infected cells (sh-ctrl) were treated for 12 hours with apigenin (20 µM), ABT-263 (0.5 µM) or their combination. The cells were harvested for cell death analysis (upper panel) or western blotting analysis of Mcl-1 and cleavage of caspase 3 (lower panel).

Article Snippet: Lentiviral plasmid of pCDH1-HA-Mcl-1 and its control pCDH1-EF1-puro were gifts of Dr. Shengbing Huang (Mayo Clinic, MN) Cell lines The human colon cancer cell lines DLD1, HCT116, HCT-8, HT29 and SW48 were obtained from American Type Culture Collection (Manassas, VA).

Techniques: Expressing, Western Blot, shRNA, Knockdown, Control, Infection

Journal: Molecular cancer therapeutics

Article Title: Apigenin sensitizes colon cancer cells to anti-tumor activity of ABT-263

doi: 10.1158/1535-7163.MCT-13-0066

Figure Lengend Snippet: A, HCT116 and DLD1 cells were treated with increasing concentrations of apigenin (apg) for 24 hours. The effect of treatments on phospho-AKT (S473) was examined by western blotting. B, The levels of phospho-AKT were analyzed in HCT116 and DLD1 cells treated for 1 day with ABT-263 (ABT) (0.5 µM), apigenin (20 µM) or their combination. C, AKT1 and AKT2 in HCT116 cells were knocked down individually or simultaneously by shRNA and knockdown efficiencies were confirmed by western blotting (left). The cells were treated for 1 day with ABT-263 followed by cell death analysis (right). D, HCT116 and DLD1 cells were treated for 1 day with ABT-263, MK2206 (MK) or their combination (comb) at the indicated concentrations followed by analysis of cell death. E, the cells were treated as described in D and subjected to western blotting analysis of phospho-AKT and cleaved caspase 3.

Article Snippet: Lentiviral plasmid of pCDH1-HA-Mcl-1 and its control pCDH1-EF1-puro were gifts of Dr. Shengbing Huang (Mayo Clinic, MN) Cell lines The human colon cancer cell lines DLD1, HCT116, HCT-8, HT29 and SW48 were obtained from American Type Culture Collection (Manassas, VA).

Techniques: Western Blot, shRNA, Knockdown

Journal: Molecular cancer therapeutics

Article Title: Apigenin sensitizes colon cancer cells to anti-tumor activity of ABT-263

doi: 10.1158/1535-7163.MCT-13-0066

Figure Lengend Snippet: A, HCT116 and DLD1 cells were treated for 1 day with the indicated concentrations of apigenin and analyzed by western blotting for phospho-ERK1/2 (T202/Y204) and total ERK1/2. B, HCT116 and DLD1 cells were treated for 1 day with ABT-263 (0.5 µM) apigenin (20 µM), or their combination before western blotting analysis phospho-ERK1/2 (T202/Y204) and ERK1/2 as in A. C, ERK1 and ERK2 in HCT116 and DLD1 cells were knocked down individually or simultaneously by shRNA and the knockdown efficiencies were confirmed by western blotting (left). The control cells and ERK knockdown cells were treated for 1 day with ABT-263 and analyzed for cell death rates (right). D, HCT116 and DLD1 cells were treated with ABT-263, AZD6244 (AZD) or their combination (comb) at the indicated concentrations for 1 day before determination of cell death as in C. E, The cells were treated as described in D and subjected to western blotting analysis of phospho-ERK1/2 and cleaved caspase 3.

Article Snippet: Lentiviral plasmid of pCDH1-HA-Mcl-1 and its control pCDH1-EF1-puro were gifts of Dr. Shengbing Huang (Mayo Clinic, MN) Cell lines The human colon cancer cell lines DLD1, HCT116, HCT-8, HT29 and SW48 were obtained from American Type Culture Collection (Manassas, VA).

Techniques: Western Blot, shRNA, Knockdown, Control

Journal: Advanced Science

Article Title: Single‐Cell Transcriptomics Unravels Growth Factor Erv1‐Like Mediated Ferroptosis as a Key Driver of Intestinal Epithelial Dysfunction in Ulcerative Colitis

doi: 10.1002/advs.202502014

Figure Lengend Snippet: In UC, enterocyte cells are primarily associated with ferroptosis. A) Heatmap of GSVA scores for 15 modes of cell death in colonic epithelial cells. B) Dot plot of AddModuleScore for 15 modes of cell death in colonic epithelial cells. C) The violin plot of the AUCell score for ferroptosis in colonic epithelial cells. D) The UMAP plot of the AUCell score for ferroptosis in colonic epithelial cells. E) The H&E staining of the clinical tissues. F) The PAS staining of the clinical tissues. G) Ferrous iron content of clinical tissues. H) Relative mRNA expression of FTH1 in clinical tissues. I) Relative mRNA expression of ACSL4 in clinical tissues. J) Relative mRNA expression of GPX4 in clinical tissues. K) Transmission electron micrographs of the clinical tissues. L) Relative mRNA expression levels of Acsl4 , Fth1 , and Gpx4 from animal tissues. M) Relative mRNA expression levels of ACSL4 , FTH1 , and GPX4 in FHC cells after Fer‐1 administration. N) Protein expression levels of ACSL4, FTH1, and GPX4 from animal tissues. O) Protein expression levels of ACSL4, FTH1, and GPX4 in FHC cells after Fer‐1 administration.

Article Snippet: The FHC colon epithelial cell line was used, FHC cells were purchased from the American Type Culture Collection (#CRL‐1831, ATCC, Manassas, VA, USA).

Techniques: Staining, Expressing, Transmission Assay

Journal: Advanced Science

Article Title: Single‐Cell Transcriptomics Unravels Growth Factor Erv1‐Like Mediated Ferroptosis as a Key Driver of Intestinal Epithelial Dysfunction in Ulcerative Colitis

doi: 10.1002/advs.202502014

Figure Lengend Snippet: GFER protects against ferroptosis in colonic epithelial cells in an inflammation model. A) Relative mRNA expression level of GFER in FHC cells after GFER overexpression. B) Protein expression level of GFER in FHC cells after GFER overexpression. C) GSH content in FHC cells after GFER overexpression. D) MDA content in FHC cells after GFER overexpression. E) Relative mRNA expression level of ACSL4 in FHC cells after GFER overexpression. F) Relative mRNA expression level of FTH1 in FHC cells after GFER overexpression. G) Relative mRNA expression level of GPX4 in FHC cells after GFER overexpression. H) Protein expression levels of ACSL4, FTH1, and GPX4 in FHC cells after GFER overexpression. I) ROS content in FHC cells after GFER overexpression. J) Superoxide anion content in FHC cells after GFER overexpression. K) Transmission electron microscopy images of FHC cells after GFER overexpression.

Article Snippet: The FHC colon epithelial cell line was used, FHC cells were purchased from the American Type Culture Collection (#CRL‐1831, ATCC, Manassas, VA, USA).

Techniques: Expressing, Over Expression, Transmission Assay, Electron Microscopy