Journal: Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology

Article Title: Plasminogen Activator Inhibitor 1 Promotes Immunosuppression in Human Non-Small Cell Lung Cancers by Enhancing TGF-Β1 Expression in Macrophage.

doi: 10.1159/000486025

Figure Lengend Snippet: Fig. 4. TGF-β promotion effect of PAI-1 was via a TLR4 / NF-ΚB dependent pathway. A) Phosphorylated IκBα at S54 as well as total IκBα were assessed using western blot in THP-1- and U937-derived macrophages treated with varying PAI-1 concentrations. B) Wild-type and TLR4 knockdown THP-1- and U937-derived macrophages were treated with varying PAI-1 concentrations and phosphorylated IκBα at S54, total IκBα and TLR4 levels were assessed by western blot. C) IL-6 levels from THP-1- and U937-derived macrophages treated with PAI-1 and Bay 11-7082 were determined by western blot. D) TGF-b levels, phosphorylated IκBα at S54, as well as total IκBα were assessed using western blot in THP-1-derived macrophages treated with PAI-1, Bay 11-7082 and BP-1-102.

Article Snippet: Published by S. Karger AG, Basel www.karger.com/cpb Zhu et al.: PAI-1 TGF-B1 Freeforward Loop in NSCLCs TAMs TLR4 was purchased from Santa Cruz (sc-40260-SH, CA) and the expression of TLR4 was knocked down per the manufacturer’s instructions.

Techniques: Western Blot, Derivative Assay, Knockdown

Journal: Breast Cancer: Targets and Therapy

Article Title: Identification of genes involved in breast cancer and breast cancer stem cells

doi: 10.2147/bctt.s85202

Figure Lengend Snippet: Figure 2 Gene expression among different cancer types. Notes: Relative gene expression analysis of nine genes that were overexpressed in breast cancer and breast CSCs. The gene expression also was studied in colon and lung cancer, colon CSCs, lung CSCs, embryonic stem cells, and in a reference sample. The experiments were performed in triplicate and a P-value of ,0.05 was considered significant. Abbreviations: CSCs, cancer stem cells; ESCs, embryonic stem cells; HP, haptoglobin; SMPD1, sphingomyelin phosphodiesterase 1, acid lysosomal; SCRIB, scribbled homolog (Drosophila); KCMF1, potassium channel modulatory factor 1; FAM155B, family with sequence similarity 155, member B; PTGER3, prostaglandin E receptor 3 (subtype EP3); GPR3, G protein-coupled receptor 3; TMX2, thioredoxin-related transmembrane protein 2; DDX49, DEAD (Asp-Glu-Ala-Asp) box polypeptide 49.

Article Snippet: Human breast CSCs (36102-29P; Celprogen, CA, USA), colon CSCs (36112-39P; Celprogen), and lung CSCs (36107-34P; Celprogen) were cultured in appropriate growth medium provided by the manufacturer (M36102-29E, M36112-39E, M36107-34E; Celprogen) in 25 cm2 flasks (E36102-29-T25, E36112-39-T25, E36107-34-T25; Celprogen).

Techniques: Gene Expression, Sequencing

Journal: Anatomical record (Hoboken, N.J. : 2007)

Article Title: Alteration in metastasis potential and gene expression in human lung cancer cell lines by ITGB8 silencing.

doi: 10.1002/ar.22521

Figure Lengend Snippet: Fig. 1. Expression of ITGB8 in different stably transfected clones of human lung cancer cell lines. The total cell lysate was separated by SDS-PAGE gel followed by Western blot with anti-ITGB8 antibody or anti-b-actin antibody. (A) A549 parent cell, A549 negative control cell line, and A549-ITGB8 silencing cell line. b-actin was used as an inter- nal control for loading. (B) PC9 parent cell, PC9 negative control cell line, and PC9-ITGB8 silencing cell line. b-actin was used as an internal control for loading. The experiment shown is representative of three independent experiments with similar results. (C) Expression of ITGB8 mRNA in different stably transfected clones of human lung cancer cell lines. Bars indicate SD. N ¼ 3. #P > 0.05; **P < 0.05 compared with the parent group.

Article Snippet: An empty vector and ITGB8 shRNA plasmid (Santa Cruz biotechnology) were transfected into human lung cancer cell lines A549 and PC9 with Lipofectamine 2000 reagent.

Techniques: Expressing, Stable Transfection, Transfection, Clone Assay, SDS Page, Western Blot, Negative Control, Control

Journal: Anatomical record (Hoboken, N.J. : 2007)

Article Title: Alteration in metastasis potential and gene expression in human lung cancer cell lines by ITGB8 silencing.

doi: 10.1002/ar.22521

Figure Lengend Snippet: Fig. 7. Effect of ITGB8 silencing on CXCL1, 2, 5 secretions in human lung cancer cell lines. (A) Effect of ITGB8 silencing on the CXCL1 secretion in A549 and PC9 cells. Bars indicate SD. n ¼ 3. #P > 0.05; **P < 0.05 compared with the parent group. (B) Effect of ITGB8 silencing on the CXCL2 secretion in A549 and PC9 cells. Bars indicate SD. n ¼ 3. #P > 0.05; **P < 0.05 compared with the parent group. (C) Effect of ITGB8 silencing on the CXCL5 secretion in A549 and PC9 cells. Bars indicate SD. n ¼ 3. #P > 0.05; **P < 0.05 com- pared with the parent group.

Article Snippet: An empty vector and ITGB8 shRNA plasmid (Santa Cruz biotechnology) were transfected into human lung cancer cell lines A549 and PC9 with Lipofectamine 2000 reagent.

Techniques:

Journal: Anatomical record (Hoboken, N.J. : 2007)

Article Title: Alteration in metastasis potential and gene expression in human lung cancer cell lines by ITGB8 silencing.

doi: 10.1002/ar.22521

Figure Lengend Snippet: Fig. 8. Effect of ITGB8 silencing on p-Akt expression in human lung cancer cell lines. The total cell lysate was separated by SDS-PAGE gel followed by Western blot with anti-p-Akt antibody or anti-b-actin antibody. (A) A549 parent cell, A549 negative control cell line, and A549-ITGB8-silencing cell line. b-actin was used as an internal control for loading. (B) PC9 parent cell, PC9 negative control cell line, and PC9 ITGB8 silencing cell line. b-actin was used as an internal control for loading. The experiment shown is representative of three inde- pendent experiments with similar results.

Article Snippet: An empty vector and ITGB8 shRNA plasmid (Santa Cruz biotechnology) were transfected into human lung cancer cell lines A549 and PC9 with Lipofectamine 2000 reagent.

Techniques: Expressing, SDS Page, Western Blot, Negative Control, Control

Journal: Anatomical record (Hoboken, N.J. : 2007)

Article Title: Alteration in metastasis potential and gene expression in human lung cancer cell lines by ITGB8 silencing.

doi: 10.1002/ar.22521

Figure Lengend Snippet: Fig. 9. Effect of ITGB8 silencing on p-MEK expression in human lung cancer cell lines. The total cell lysate was separated by SDS- PAGE gel followed by Western blot with anti-p-MEK antibody or anti- b-actin antibody. (A) A549 parent cell, A549 negative control cell line, and A549-ITGB8 silencing cell line. b-actin was used as an internal control for loading. (B) PC9 parent cell, PC9 negative control cell line, and PC9-ITGB8 silencing cell line. b-actin was used as an internal control for loading. The experiment shown is representative of three independent experiments with similar results.

Article Snippet: An empty vector and ITGB8 shRNA plasmid (Santa Cruz biotechnology) were transfected into human lung cancer cell lines A549 and PC9 with Lipofectamine 2000 reagent.

Techniques: Expressing, SDS Page, Western Blot, Negative Control, Control

Journal: BMC cancer

Article Title: TRPM7 overexpression enhances the cancer stem cell-like and metastatic phenotypes of lung cancer through modulation of the Hsp90α/uPA/MMP2 signaling pathway.

doi: 10.1186/s12885-018-5050-x

Figure Lengend Snippet: Fig. 3 Downregulation of TRPM7 expression suppress the viability and metastatic phenotype of human lung cancer cells. a The effect of shTRPM7 clones 1 and 2 on the mRNA expression levels of TRPM7, Vimentin, E-cadherin, p21 and BAK. b Fewer colonies were formed by the shTRPM7 clones compared to the control A549 and 95D cells. c Cell viability of A549 and 95D cells is significantly reduced in shTRPM7 clones, compared to the control cells. d shTRPM7 clones exhibited suppressed migration and invasion potential, in comparison to the control A549 and 95D cells. GAPDH served as loading control. * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: Human lung cancer cells were fixed in 10% paraformaldehyde for 30 min, blocked with goat serum for 30min, then incubated with anti-TRPM7 and anti-SOX2 antibodies (Santa Cruz Biotechnology) at a dilution of 1:100 for 1 h at 37 °C.

Techniques: Expressing, Clone Assay, Control, Migration, Comparison

Journal: BMC cancer

Article Title: TRPM7 overexpression enhances the cancer stem cell-like and metastatic phenotypes of lung cancer through modulation of the Hsp90α/uPA/MMP2 signaling pathway.

doi: 10.1186/s12885-018-5050-x

Figure Lengend Snippet: Fig. 4 TRPM7 regulates the CSCs activities of lung cancer cells by modulating the Hsp90α/uPA/MMP2 signaling pathway. a Representative RT-PCR ananylsis showing upregulated TRPM7, SOX2, CD133, KLF4, HSP90α, uPA, and MMP2 in 95D tumorspheres, compared to parental 95D cells. b Photo images showing shTRPM7 clones lost ability to form tumorspheres in comparison to the control 95D cells, upper panel; TRPM7, HSP90α, uPA, and MMP2 mRNA expression is significantly downregulated in the tumorspheres derived from shTRPM7 clones, lower panel. c Immunoflorescence staining showing shTRPM7 95D tumorspheres were significantly smaller, compared to the control 95D tumorspheres. d Identification of side population (SP) cells from the control or shTRPM7 95D cells. SP cells were sorted by flow cytometry using Hoechst red and blue from 95D cells with or without verapamil pretreatment for 30 min. GAPDH served as loading control. * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: Human lung cancer cells were fixed in 10% paraformaldehyde for 30 min, blocked with goat serum for 30min, then incubated with anti-TRPM7 and anti-SOX2 antibodies (Santa Cruz Biotechnology) at a dilution of 1:100 for 1 h at 37 °C.

Techniques: Reverse Transcription Polymerase Chain Reaction, Clone Assay, Comparison, Control, Expressing, Derivative Assay, Staining, Flow Cytometry

Journal: BMC cancer

Article Title: TRPM7 overexpression enhances the cancer stem cell-like and metastatic phenotypes of lung cancer through modulation of the Hsp90α/uPA/MMP2 signaling pathway.

doi: 10.1186/s12885-018-5050-x

Figure Lengend Snippet: Fig. 5 TRPM7 inhibitor, Waixenicin A suppresses the CSC phenotype of lung cancer cells. a Chemical structure of Waixenicin A with molecular formular C26H36O7 and molecular weight 460.56 mg/mol. b The inhibitory effect of 5 μM and 10 μM Waixenicin A on the expression levels of TRPM7, Vimentin, Survivin, STAT3, HSP90α, uPA, and MMP2 in 95D cells. c Photo image and (d) histography showing significantly inhibited tumorsphere formation by shTRPM7 and/or Waixenicin A treatment

Article Snippet: Human lung cancer cells were fixed in 10% paraformaldehyde for 30 min, blocked with goat serum for 30min, then incubated with anti-TRPM7 and anti-SOX2 antibodies (Santa Cruz Biotechnology) at a dilution of 1:100 for 1 h at 37 °C.

Techniques: Molecular Weight, Expressing

Journal: BMC cancer

Article Title: TRPM7 overexpression enhances the cancer stem cell-like and metastatic phenotypes of lung cancer through modulation of the Hsp90α/uPA/MMP2 signaling pathway.

doi: 10.1186/s12885-018-5050-x

Figure Lengend Snippet: Fig. 6 Pictorial Abstract showing that the molecular or therapeutic inhibition of TRPM7 expression and/or activity attenuates the cancer stem cell-like and metastatic phenotypes of lung cancer cells through the modulation of the Hsp90α/uPA/MMP2 signaling

Article Snippet: Human lung cancer cells were fixed in 10% paraformaldehyde for 30 min, blocked with goat serum for 30min, then incubated with anti-TRPM7 and anti-SOX2 antibodies (Santa Cruz Biotechnology) at a dilution of 1:100 for 1 h at 37 °C.

Techniques: Inhibition, Expressing, Activity Assay