Journal: Oncotarget

Article Title: Cancer cell-selective, clathrin-mediated endocytosis of aptamer decorated nanoparticles

doi: 10.18632/oncotarget.24772

Figure Lengend Snippet: Cells were incubated with 50 nM S15-APT QDs for 2 h. Nuclei were stained with 2 μg/ml Hoechst 33342. Fluorescence confocal microscopy was performed using an inverted confocal microscope (Zeiss LSM 710) at ×630 magnification. ( A ) Human A549 non-small cell lung carcinoma cells; ( B ) Normal human bronchial epithelial BEAS2B cells which served as normal non-target cells, ( C ) Human colon adenocarcinoma CaCo-2 cells; ( D ) Human cervical carcinoma HeLa cells; ( E ) A549 cells were incubated with 50 nM S15-APT QDs along with 5 μM free APT; ( F ) A549 cells were incubated with 50 nM random sequence APT-QDs; ( G ) A549 cells incubated with no S15-APT QDs; H) A549 cells incubated with 50 nM Qdot ® 655; ( I ) ABCG2-overexpressing MDR subline A549/K1.5 cells incubated with 50 nM S15-APT QDs; the red fluorescence channel was defined between 10-100 for all presented images.

Article Snippet: Normal human bronchial epithelial BEAS2B cells were generously provided by Prof. Rotem Karni (The Hebrew University, Jerusalem, Israel).

Techniques: Incubation, Staining, Fluorescence, Confocal Microscopy, Microscopy, Sequencing

Journal: Toxicology and applied pharmacology

Article Title: Upregulation of histone-lysine methyltransferases plays a causal role in hexavalent chromium-induced cancer stem cell-like property and cell transformation

doi: 10.1016/j.taap.2018.01.022

Figure Lengend Snippet: (A,B) Quantitation of chronic low dose Cr(VI) exposure-induced soft agar colony and suspension culture sphere formation by human bronchial epithelial cells (BEAS-2B and 16HBE). After exposure to 0.25 μM of Cr(VI) (K2Cr2O7) for 20 (BEAS-2B) and 40 (16HBE) weeks, passage-matched control and Cr(VI)-exposed BEAS-2B and 16 HBE cells were harvested for soft agar colony formation (A) and suspension culture spheroid formation (B) assay as described in Methods. The results are presented as means ± standard deviations (n=3). *p<0.05, compared to passage-matched vehicle (H2O) control-exposed cells. (C) Representative Western blot images for the analysis of histone H3 posttranslational modifications in passage- matched control and Cr(VI)-exposed BEAS-2B and 16 HBE cells. After exposure to 0.25 μM of Cr(VI) (K2Cr2O7) for 20 (BEAS-2B) and 40 (16HBE) weeks, passage-matched control and Cr(VI)-exposed BEAS-2B and 16 HBE cells were harvested for Western blot analysis as described in Methods. Similar results were obtained in two additional experiments.

Article Snippet: Passage-matched control and Cr(VI)-transformed BEAS-2B cells (1.5 × 10 6 cells in 0.1 ml of 1:1 growth factor-reduced matrigel and PBS) were injected subcutaneously into the right flank of female nude mice (Nu/Nu, Charles River laboratories, four mice in each group).

Techniques: Quantitation Assay, Western Blot

Journal: Toxicology and applied pharmacology

Article Title: Upregulation of histone-lysine methyltransferases plays a causal role in hexavalent chromium-induced cancer stem cell-like property and cell transformation

doi: 10.1016/j.taap.2018.01.022

Figure Lengend Snippet: (A) Representative Western blot images for the analysis of several HMTases levels in passage-matched control and Cr(VI)-exposed human bronchial epithelial. After exposure to 0.25 μM of Cr(VI) (K2Cr2O7) for 20 (BEAS-2B), 40 (16HBE) and 30 (HBEC3-KT) weeks, passage- matched control and Cr(VI)-exposed BEAS-2B, 16 HBE and HBEC3-KT cells were harvested for Western blot analysis as described in Methods. Similar results were obtained in two additional experiments. (B) Representative images of H&E staining and immunofluorescence (IF) staining of SUV39H1 and EZH2 in the squamous cell lung carcinoma tissue and the adjacent normal lung tissue from a non-smoker worker exposed to chromate for 19 years. Similar staining results were obtained in lung cancer tissue from another non-smoker worker exposed chromate for 38 years. Scale bar: 100 μm.

Article Snippet: Passage-matched control and Cr(VI)-transformed BEAS-2B cells (1.5 × 10 6 cells in 0.1 ml of 1:1 growth factor-reduced matrigel and PBS) were injected subcutaneously into the right flank of female nude mice (Nu/Nu, Charles River laboratories, four mice in each group).

Techniques: Western Blot, Staining, Immunofluorescence

Journal: Toxicology and applied pharmacology

Article Title: Upregulation of histone-lysine methyltransferases plays a causal role in hexavalent chromium-induced cancer stem cell-like property and cell transformation

doi: 10.1016/j.taap.2018.01.022

Figure Lengend Snippet: (A) Representative Western blot images for the analysis of histone H3 repressive methylation marks in Cr(VI)-transformed cells. Cells were treated with vehicle control, a G9a inhibitor BIX01294 (2.5 μM) or an EZH2 inhibitor DZNeP (0.25 μM) for 72 h, and harvested for Western blot analysis. (B) Quantitation of effects of inhibition of G9a or EZH2 on soft agar colony and suspension culture sphere formation by Cr(VI)-transformed cells. Cells were treated with vehicle control, a G9a inhibitor BIX01294 (2.5 μM) or an EZH2 inhibitor DZNeP (0.25 μM) for 72 h, and harvested for soft agar colony and suspension culture sphere formation assay. Results are expressed as relative colony or sphere formation compared to vehicle-treated BEAS-2B-Cr(VI) cells (100%). Data are presented as mean ± SD (n=3). *p< 0.05, compared to vehicle-treated group. Similar results were obtained in two additional experiments.

Article Snippet: Passage-matched control and Cr(VI)-transformed BEAS-2B cells (1.5 × 10 6 cells in 0.1 ml of 1:1 growth factor-reduced matrigel and PBS) were injected subcutaneously into the right flank of female nude mice (Nu/Nu, Charles River laboratories, four mice in each group).

Techniques: Western Blot, Methylation, Transformation Assay, Quantitation Assay, Inhibition, Tube Formation Assay

Journal: Toxicology and applied pharmacology

Article Title: Upregulation of histone-lysine methyltransferases plays a causal role in hexavalent chromium-induced cancer stem cell-like property and cell transformation

doi: 10.1016/j.taap.2018.01.022

Figure Lengend Snippet: (A) Representative Western blot images for the analysis of levels of HMTases in G9a, SUV391 and EZH2 shRNA stable knockdown BEAS-2B cells. Generation of shRNA vector control and each HMTase shRNA stable knockdown cells are described in Methods. (B) Measurement of intracellular chromium levels. Cells were treated with 10 μM of K2Cr2O7 for 3 h and collected for intracellular chromium level analysis as described in Methods. Data are presented as mean ± SD (n=3). (C,D) Quantitation of effects of stable knockdown of G9a, SUV39H1 or EZH2 on chronic Cr(VI) exposure-induced soft agar colony (C) and suspension culture sphere formation (D). Vector control and each HMTase stable knockdown BEAS-2B cells were exposed to 0.125 μM of Cr(VI) (K2Cr2O7) for 25 weeks as described in Methods. At the end of Cr(VI) exposure, cells were collected for soft agar colony and suspension culture sphere formation assays. Results are expressed as relative colony or sphere formation compared to Cr(VI)-exposed shRNA vector control cells (100%). Data are presented as mean ± SD (n=3). *p< 0.05, compared to Cr(VI)-exposed shRNA vector control cells. Similar results were obtained in two additional experiments.

Article Snippet: Passage-matched control and Cr(VI)-transformed BEAS-2B cells (1.5 × 10 6 cells in 0.1 ml of 1:1 growth factor-reduced matrigel and PBS) were injected subcutaneously into the right flank of female nude mice (Nu/Nu, Charles River laboratories, four mice in each group).

Techniques: Western Blot, shRNA, Plasmid Preparation, Quantitation Assay

Journal: Toxicology and applied pharmacology

Article Title: Upregulation of histone-lysine methyltransferases plays a causal role in hexavalent chromium-induced cancer stem cell-like property and cell transformation

doi: 10.1016/j.taap.2018.01.022

Figure Lengend Snippet: (A) Representative images of immunofluorescence (IF) staining of γH2AX in chronic low dose Cr(VI)-exposed shRNA vector control and each HMTase shRNA stable knockdown BEAS-2B cells treated with 2.5 μM of Cr(VI) (K2Cr2O7). After exposure to 0.125 μM of Cr(VI) (K2Cr2O7) for 25 weeks, vector control and each HMTase stable knockdown cells were seeded in 6-well plates. After overnight culture, cells were treated with 2.5 μM of K 2Cr2O7 for 12 h. At the end of the treatment, one set of cells were used for IF staining of γH2AX (designated as Rest 0 h). The other set of cells were washed with PBS and cultured for additional 36 h in the absence of K2Cr2O7 and used for IF staining of γH2AX (designated as Rest 36 h) as described in Methods. Scale bar: 100 μm. (B) Quantitation of γH2AX positive staining in Cr(VI)-exposed cells described in (A). Results are expressed as percent (%) of γH2AX positive staining per field of view. Data are presented as mean ± SD (n=30 fields of view). *p< 0.05, compared to Cr(VI)-exposed shRNA vector control cells. Similar results were obtained in two additional experiments.

Article Snippet: Passage-matched control and Cr(VI)-transformed BEAS-2B cells (1.5 × 10 6 cells in 0.1 ml of 1:1 growth factor-reduced matrigel and PBS) were injected subcutaneously into the right flank of female nude mice (Nu/Nu, Charles River laboratories, four mice in each group).

Techniques: Immunofluorescence, Staining, shRNA, Plasmid Preparation, Cell Culture, Quantitation Assay