Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Furanodiene Induces Endoplasmic Reticulum Stress and Presents Antiproliferative Activities in Lung Cancer Cells

doi: 10.1155/2012/426521

Figure Lengend Snippet: FUR inhibition on colony formation in lung cancer cells. (a) Cells were treated with various concentrations of FUR for 24 h, after which morphological changes were observed using an AxioCam HRC CCD phase contrast microscope. Bar: 20 μ m. (b) Cells were treated with various concentrations of FUR for 24 h. After treatment, cells were suspended and reseeded into 6-well plates at a density of 200 cells per well (A549) or 1000 cells per well (95-D) and then fixed and stained with 4% PFA and crystal violet after one week. The representative images of colony formation assay were obtained. Bar: 2 mm.

Article Snippet: The human lung cancer cell lines A549 (ATCC, Rockville, MD, USA), NCI-H1299 (ATCC), and 95-D (The Cell Bank of Type Culture Collection of Chinese Academy of Sciences, Shanghai, China) were cultured in RPMI 1640 medium supplemented with 10% fetal bovine serum (GIBCO BRL, Carlsbad, CA, USA), 100 U/mL penicillin, and 100 mg/mL streptomycin and were grown in an incubator with 5% CO 2 at 37°C.

Techniques: Inhibition, Microscopy, Staining, Colony Assay

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Furanodiene Induces Endoplasmic Reticulum Stress and Presents Antiproliferative Activities in Lung Cancer Cells

doi: 10.1155/2012/426521

Figure Lengend Snippet: FUR induces ER stress in lung cancer cells. (a) Effect of FUR on the mRNA levels of BIP and CHOP in A549 and 95-D cells. Cells were treated with different concentrations of FUR for 12 h, and mRNA levels were determined by RT-PCR. (b) Effect of FUR on the protein levels of BIP and CHOP in A549 and 95-D cells. Cells were treated with different concentrations of FUR for 12 h or 24 h, and protein levels were determined by Western blot. (c) Immunocytochemical staining was conducted to detect the expression of CHOP in nuclei. Cells were treated with 80 μ M FUR for 12 h and the cells were fixed and stained with anti-CHOP antibody (green) and Hoechst 33342 (blue). Bar: 10 μ m.

Article Snippet: The human lung cancer cell lines A549 (ATCC, Rockville, MD, USA), NCI-H1299 (ATCC), and 95-D (The Cell Bank of Type Culture Collection of Chinese Academy of Sciences, Shanghai, China) were cultured in RPMI 1640 medium supplemented with 10% fetal bovine serum (GIBCO BRL, Carlsbad, CA, USA), 100 U/mL penicillin, and 100 mg/mL streptomycin and were grown in an incubator with 5% CO 2 at 37°C.

Techniques: Reverse Transcription Polymerase Chain Reaction, Western Blot, Staining, Expressing

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Furanodiene Induces Endoplasmic Reticulum Stress and Presents Antiproliferative Activities in Lung Cancer Cells

doi: 10.1155/2012/426521

Figure Lengend Snippet: CI values of FUR at concentrations applied in combination with DOX or TAX in different lung cancer cell lines.

Article Snippet: The human lung cancer cell lines A549 (ATCC, Rockville, MD, USA), NCI-H1299 (ATCC), and 95-D (The Cell Bank of Type Culture Collection of Chinese Academy of Sciences, Shanghai, China) were cultured in RPMI 1640 medium supplemented with 10% fetal bovine serum (GIBCO BRL, Carlsbad, CA, USA), 100 U/mL penicillin, and 100 mg/mL streptomycin and were grown in an incubator with 5% CO 2 at 37°C.

Techniques:

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