Journal: Molecular medicine reports

Article Title: S100A16 suppresses the proliferation, migration and invasion of colorectal cancer cells in part via the JNK/p38 MAPK pathway.

doi: 10.3892/mmr.2020.11803

Figure Lengend Snippet: Figure 1. S100A16 is decreased in human colorectal cancer. (A)Analysis of S100A16 expression in normal colorectal tissues and colorectal adenocarcinoma tissues in the Skrzypczak in Oncomine microarray dataset, as assessed using an unpaired-t test. P<0.05. (B) Representative images of S100A16 protein expres sion in CRC tumour tissues and paired normal adjacent tissues, as assessed via immunohistochemistry. Magnification, x400. **P<0.01, paired Student's t-test. (C) Representative images of difference scores indicating S100A16 protein expression in CRC tumour tissues and paired normal adjacent tissues (magnifica tion, x400). The proportion of S100A16-expressing in CRC tissue samples was also analysed. **P<0.01, χ2 test. S100A16 (D) mRNA and (E) protein expression in CRC cell lines, as determined via reverse transcription-quantitative PCR and western blotting, respectively. (F) Kaplan-Meier analysis of the survival rates in patients with CRC with S100A16 low or high expression. CRC, colorectal cancer; S100A16, S100 calcium binding protein A16, IRS, immunoreactive score.

Article Snippet: After being blocked with non-fat milk for 1 h at room temperature, the membranes were incubated with the following primary antibodies at 4 ̊C overnight: anti-rabbit S100A4 (cat. no. 16105-1-AP; 1:500; ProteinTech Group, Inc.), anti-rabbit S100A14 (cat. no. 10489-1-AP; 1:500; ProteinTech Group, Inc.), anti-rabbit S100A16 (cat. no. 11456-1-AP; 1:500; ProteinTech Group, Inc.) anti-phosphorylated (p)-rabbit JNK (cat. no. 9255; Cell Signaling Technology, Inc.), anti-rabbit JNK (cat. no. 9252; Cell Signaling Technology, Inc.), anti-prabbit ERK1/2 (cat. no. 4370; Cell Signaling Technology, Inc.), anti-rabbit ERK1/2 (cat. no. 4695; Cell Signaling Technology, Inc.), anti-p-rabbit p38 MAPK (cat. no. 9216; Cell Signaling Technology, Inc.), anti-rabbit p38 MAPK (cat. no. 9212; Cell Signaling Technology, Inc.), anti-rabbit vimentin (cat. no. 12826; Cell Signaling Technology, Inc.), anti-mouse E-cadherin (cat. no. 14472; Cell Signaling Technology, Inc.), anti-mouse N-cadherin (cat. no. 14215; Cell Signaling Technology, Inc.) and anti-mouse GAPDH (cat. no. 51332; Cell Signaling Technology, Inc.).

Techniques: Expressing, Microarray, Immunohistochemistry, Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot, Binding Assay

Journal: Molecular medicine reports

Article Title: S100A16 suppresses the proliferation, migration and invasion of colorectal cancer cells in part via the JNK/p38 MAPK pathway.

doi: 10.3892/mmr.2020.11803

Figure Lengend Snippet: Figure 2. S100A16 inhibits the proliferation of CRC cells. (A) Western blot analysis of S100A16 protein expression in CRC cells transfected with S100A16 siRNAs compared with si-Ctrl. Data were analysed using a one-way ANOVA followed by Tukey's post hoc test. **P<0.01. (B) Western blot analysis of S100A16 protein expression in CRC cells transfected with overexpression plasmids compared vectors, as assessed using an unpaired Student's t-test. **P<0.01. (C) S100A16 knockdown promoted the proliferation of HCT116 and SW480 cells. *P<0.05, vs. si-Ctrl. (D) S100A16 overexpression suppressed the proliferation of Lovo cells. *P<0.05. Cell proliferation was examined by performing Cell Counting Kit-8 assays and analysed using a two-way ANOVA with Bonferronis correction. CRC, colorectal cancer; siRNA/si, small interfering RNA; Ctrl, control; S100A16, S100 calcium binding protein A16; OD, optical density.

Article Snippet: After being blocked with non-fat milk for 1 h at room temperature, the membranes were incubated with the following primary antibodies at 4 ̊C overnight: anti-rabbit S100A4 (cat. no. 16105-1-AP; 1:500; ProteinTech Group, Inc.), anti-rabbit S100A14 (cat. no. 10489-1-AP; 1:500; ProteinTech Group, Inc.), anti-rabbit S100A16 (cat. no. 11456-1-AP; 1:500; ProteinTech Group, Inc.) anti-phosphorylated (p)-rabbit JNK (cat. no. 9255; Cell Signaling Technology, Inc.), anti-rabbit JNK (cat. no. 9252; Cell Signaling Technology, Inc.), anti-prabbit ERK1/2 (cat. no. 4370; Cell Signaling Technology, Inc.), anti-rabbit ERK1/2 (cat. no. 4695; Cell Signaling Technology, Inc.), anti-p-rabbit p38 MAPK (cat. no. 9216; Cell Signaling Technology, Inc.), anti-rabbit p38 MAPK (cat. no. 9212; Cell Signaling Technology, Inc.), anti-rabbit vimentin (cat. no. 12826; Cell Signaling Technology, Inc.), anti-mouse E-cadherin (cat. no. 14472; Cell Signaling Technology, Inc.), anti-mouse N-cadherin (cat. no. 14215; Cell Signaling Technology, Inc.) and anti-mouse GAPDH (cat. no. 51332; Cell Signaling Technology, Inc.).

Techniques: Western Blot, Expressing, Transfection, Over Expression, Knockdown, Cell Counting, Small Interfering RNA, Control, Binding Assay

Journal: Molecular medicine reports

Article Title: S100A16 suppresses the proliferation, migration and invasion of colorectal cancer cells in part via the JNK/p38 MAPK pathway.

doi: 10.3892/mmr.2020.11803

Figure Lengend Snippet: Figure 4. S100A16 knockdown activates the JNK/p38 MAPK signalling pathway and promotes EMT. MAPK pathway-associated and EMT-associated pro teins were assessed via western blotting in HCT116 cells. Data were analysed with a one-way ANOVA followed by Tukey's post hoc test. **P<0.01; #P>0.05 (not significant). EMT, epithelial-mesenchymal transition; S100A16, S100 calcium binding protein A16; p-, phosphorylated; E-cad, E-cadherin; siRNA/si, small interfering RNA; Ctrl, control; N-cad, N-cadherin.

Article Snippet: After being blocked with non-fat milk for 1 h at room temperature, the membranes were incubated with the following primary antibodies at 4 ̊C overnight: anti-rabbit S100A4 (cat. no. 16105-1-AP; 1:500; ProteinTech Group, Inc.), anti-rabbit S100A14 (cat. no. 10489-1-AP; 1:500; ProteinTech Group, Inc.), anti-rabbit S100A16 (cat. no. 11456-1-AP; 1:500; ProteinTech Group, Inc.) anti-phosphorylated (p)-rabbit JNK (cat. no. 9255; Cell Signaling Technology, Inc.), anti-rabbit JNK (cat. no. 9252; Cell Signaling Technology, Inc.), anti-prabbit ERK1/2 (cat. no. 4370; Cell Signaling Technology, Inc.), anti-rabbit ERK1/2 (cat. no. 4695; Cell Signaling Technology, Inc.), anti-p-rabbit p38 MAPK (cat. no. 9216; Cell Signaling Technology, Inc.), anti-rabbit p38 MAPK (cat. no. 9212; Cell Signaling Technology, Inc.), anti-rabbit vimentin (cat. no. 12826; Cell Signaling Technology, Inc.), anti-mouse E-cadherin (cat. no. 14472; Cell Signaling Technology, Inc.), anti-mouse N-cadherin (cat. no. 14215; Cell Signaling Technology, Inc.) and anti-mouse GAPDH (cat. no. 51332; Cell Signaling Technology, Inc.).

Techniques: Knockdown, Western Blot, Binding Assay, Small Interfering RNA, Control

Journal: Molecular medicine reports

Article Title: S100A16 suppresses the proliferation, migration and invasion of colorectal cancer cells in part via the JNK/p38 MAPK pathway.

doi: 10.3892/mmr.2020.11803

Figure Lengend Snippet: Figure 3. S100A16 inhibits CRC cell migration and invasion. (A) S100A16 knockdown promoted the migration and invasion of the two CRC cell lines, which was analysed using a one-way ANOVA followed by Tukey's post hoc test. **P<0.01. (B) S100A16 overexpression suppressed the migration and invasion of Lovo cells, which was analysed using an unpaired Student's t-test. **P<0.01. Cell migration and invasion were determined by performing Transwell assays. CRC, colorectal cancer; S100A16, S100 calcium binding protein A16; siRNA/si, small interfering RNA; Ctrl, control.

Article Snippet: After being blocked with non-fat milk for 1 h at room temperature, the membranes were incubated with the following primary antibodies at 4 ̊C overnight: anti-rabbit S100A4 (cat. no. 16105-1-AP; 1:500; ProteinTech Group, Inc.), anti-rabbit S100A14 (cat. no. 10489-1-AP; 1:500; ProteinTech Group, Inc.), anti-rabbit S100A16 (cat. no. 11456-1-AP; 1:500; ProteinTech Group, Inc.) anti-phosphorylated (p)-rabbit JNK (cat. no. 9255; Cell Signaling Technology, Inc.), anti-rabbit JNK (cat. no. 9252; Cell Signaling Technology, Inc.), anti-prabbit ERK1/2 (cat. no. 4370; Cell Signaling Technology, Inc.), anti-rabbit ERK1/2 (cat. no. 4695; Cell Signaling Technology, Inc.), anti-p-rabbit p38 MAPK (cat. no. 9216; Cell Signaling Technology, Inc.), anti-rabbit p38 MAPK (cat. no. 9212; Cell Signaling Technology, Inc.), anti-rabbit vimentin (cat. no. 12826; Cell Signaling Technology, Inc.), anti-mouse E-cadherin (cat. no. 14472; Cell Signaling Technology, Inc.), anti-mouse N-cadherin (cat. no. 14215; Cell Signaling Technology, Inc.) and anti-mouse GAPDH (cat. no. 51332; Cell Signaling Technology, Inc.).

Techniques: Migration, Knockdown, Over Expression, Binding Assay, Small Interfering RNA, Control

Journal: Molecular medicine reports

Article Title: S100A16 suppresses the proliferation, migration and invasion of colorectal cancer cells in part via the JNK/p38 MAPK pathway.

doi: 10.3892/mmr.2020.11803

Figure Lengend Snippet: Figure 5. JNK/p38 MAPK signalling pathway inactivation is required for S100A16 knockdown-mediated HCT116 cellular effects. (A) Representative images of Transwell assays after treatment with the p38 inhibitor (SB203580) or the JNK inhibitor (SP600125) following western blotting in S100A16-silenced HCT116 cells. (B) MAPK and epithelial-mesenchymal transition markers were examined via western blotting after treatment with the p38 inhibitor (SB203580) or the JNK inhibitor (SP600125) in S100A16-silenced HCT116 cells. Data were analysed with a one-way ANOVA followed by Tukey's post hoc test. **P<0.01 and *P<0.05; #P>0.05. S100A16, S100 calcium binding protein A16; p-, phosphorylated; E-cad, E-cadherin; siRNA/si, small interfering RNA; Ctrl, control; N-cad, N-cadherin.

Article Snippet: After being blocked with non-fat milk for 1 h at room temperature, the membranes were incubated with the following primary antibodies at 4 ̊C overnight: anti-rabbit S100A4 (cat. no. 16105-1-AP; 1:500; ProteinTech Group, Inc.), anti-rabbit S100A14 (cat. no. 10489-1-AP; 1:500; ProteinTech Group, Inc.), anti-rabbit S100A16 (cat. no. 11456-1-AP; 1:500; ProteinTech Group, Inc.) anti-phosphorylated (p)-rabbit JNK (cat. no. 9255; Cell Signaling Technology, Inc.), anti-rabbit JNK (cat. no. 9252; Cell Signaling Technology, Inc.), anti-prabbit ERK1/2 (cat. no. 4370; Cell Signaling Technology, Inc.), anti-rabbit ERK1/2 (cat. no. 4695; Cell Signaling Technology, Inc.), anti-p-rabbit p38 MAPK (cat. no. 9216; Cell Signaling Technology, Inc.), anti-rabbit p38 MAPK (cat. no. 9212; Cell Signaling Technology, Inc.), anti-rabbit vimentin (cat. no. 12826; Cell Signaling Technology, Inc.), anti-mouse E-cadherin (cat. no. 14472; Cell Signaling Technology, Inc.), anti-mouse N-cadherin (cat. no. 14215; Cell Signaling Technology, Inc.) and anti-mouse GAPDH (cat. no. 51332; Cell Signaling Technology, Inc.).

Techniques: Knockdown, Western Blot, Binding Assay, Small Interfering RNA, Control

Journal: Molecular medicine reports

Article Title: S100A16 suppresses the proliferation, migration and invasion of colorectal cancer cells in part via the JNK/p38 MAPK pathway.

doi: 10.3892/mmr.2020.11803

Figure Lengend Snippet: Figure 6. Overexpression of S100A16 inhibits tumour growth in vivo. (A) Images of tumour xenografts in mice of the vector and S100A16 groups. (B) Images of the isolated tumours. (C) Tumour volume and (D) weight were separately compared between the two groups. **P<0.01. S100A16, S100 calcium binding protein A16.

Article Snippet: After being blocked with non-fat milk for 1 h at room temperature, the membranes were incubated with the following primary antibodies at 4 ̊C overnight: anti-rabbit S100A4 (cat. no. 16105-1-AP; 1:500; ProteinTech Group, Inc.), anti-rabbit S100A14 (cat. no. 10489-1-AP; 1:500; ProteinTech Group, Inc.), anti-rabbit S100A16 (cat. no. 11456-1-AP; 1:500; ProteinTech Group, Inc.) anti-phosphorylated (p)-rabbit JNK (cat. no. 9255; Cell Signaling Technology, Inc.), anti-rabbit JNK (cat. no. 9252; Cell Signaling Technology, Inc.), anti-prabbit ERK1/2 (cat. no. 4370; Cell Signaling Technology, Inc.), anti-rabbit ERK1/2 (cat. no. 4695; Cell Signaling Technology, Inc.), anti-p-rabbit p38 MAPK (cat. no. 9216; Cell Signaling Technology, Inc.), anti-rabbit p38 MAPK (cat. no. 9212; Cell Signaling Technology, Inc.), anti-rabbit vimentin (cat. no. 12826; Cell Signaling Technology, Inc.), anti-mouse E-cadherin (cat. no. 14472; Cell Signaling Technology, Inc.), anti-mouse N-cadherin (cat. no. 14215; Cell Signaling Technology, Inc.) and anti-mouse GAPDH (cat. no. 51332; Cell Signaling Technology, Inc.).

Techniques: Over Expression, In Vivo, Plasmid Preparation, Isolation, Binding Assay

Journal: Oncology reports

Article Title: Ca2+-binding protein S100A11: a novel diagnostic marker for breast carcinoma.

doi: 10.3892/or_00000764

Figure Lengend Snippet: Figure 2. Comparison of the expression of S100A11 between IDCA (B) and corresponding adjacent normal tissue (A). The arrow points to the location of the spots representing S100A11 and proteins that were up-regulated (>3-fold) in all of the comparison groups under stringent selection conditions. Visualization is by silver staining. The letter u before the number means ‘up-regulated’ and the number is the corresponding marker in each group of differential comparisons between cancer and adjacent normal tissue.

Article Snippet: After blocking with TBS containing 5% milk powder and 0.1% Tween-20 for 1 h at RT, the membranes were incubated with the anti-S100A11 antibody (ProteinTech Group, Inc.) at the concentration ratio of 1:1000 for 1 h at RT followed by treatment with horseradish peroxidase-conjugated secondary antibody for 1 h at RT.

Techniques: Comparison, Expressing, Selection, Silver Staining, Marker

Journal: Oncology reports

Article Title: Ca2+-binding protein S100A11: a novel diagnostic marker for breast carcinoma.

doi: 10.3892/or_00000764

Figure Lengend Snippet: Figure 3. S100A11 protein identified by MALDI-TOF/TOF. (A) A full scan MS-MS spectrum of protein S100A11. (B) Matched peptides are shown in panes (sequence coverage, 21%).

Article Snippet: After blocking with TBS containing 5% milk powder and 0.1% Tween-20 for 1 h at RT, the membranes were incubated with the anti-S100A11 antibody (ProteinTech Group, Inc.) at the concentration ratio of 1:1000 for 1 h at RT followed by treatment with horseradish peroxidase-conjugated secondary antibody for 1 h at RT.

Techniques: Tandem Mass Spectroscopy, Sequencing

Journal: Oncology reports

Article Title: Ca2+-binding protein S100A11: a novel diagnostic marker for breast carcinoma.

doi: 10.3892/or_00000764

Figure Lengend Snippet: Figure 4. Western blot analysis confirmed the expression of S100A11 in breast tumor tissues and adjacent normal tissues. The letter C means ‘group of cancer tissue’ and the letter N means ‘group of normal tissue’. Numbers 1-5 correspond to different subtypes (Table I). (A) Western blot for human tissues. Comparison between breast carcinoma and adjacent normal tissue. (B) Integrated density value (IDV).

Article Snippet: After blocking with TBS containing 5% milk powder and 0.1% Tween-20 for 1 h at RT, the membranes were incubated with the anti-S100A11 antibody (ProteinTech Group, Inc.) at the concentration ratio of 1:1000 for 1 h at RT followed by treatment with horseradish peroxidase-conjugated secondary antibody for 1 h at RT.

Techniques: Western Blot, Expressing, Comparison

Journal: Oncology reports

Article Title: Ca2+-binding protein S100A11: a novel diagnostic marker for breast carcinoma.

doi: 10.3892/or_00000764

Figure Lengend Snippet: Figure 5. Representative images of S100A11 immunostaining in breast tissues. Scale bar, 50 μm. (A) S100A11 positivity in breast tumor. The staining pattern is predominantly cytoplasmic (magnification, x200). (B) S100A11 positivity in corresponding adjacent normal tissue. The staining pattern is predominantly cytoplasmic (magnification, x200). (C) S100A11 positivity in breast adenosis tissue. The staining pattern is predominantly cytoplasmic (magnification, x200).

Article Snippet: After blocking with TBS containing 5% milk powder and 0.1% Tween-20 for 1 h at RT, the membranes were incubated with the anti-S100A11 antibody (ProteinTech Group, Inc.) at the concentration ratio of 1:1000 for 1 h at RT followed by treatment with horseradish peroxidase-conjugated secondary antibody for 1 h at RT.

Techniques: Immunostaining, Staining

Journal: Oncotarget

Article Title: Pancreatic cancer-derived exosomes promote tumor metastasis and liver pre-metastatic niche formation

doi: 10.18632/oncotarget.18831

Figure Lengend Snippet: Confocal microscopy (lung, liver, spleen, brain) of PKH67-labeled Panc02 EXO and Panc02-H7 EXO tissue distribution (green) 24 hpi. (A) PKH-67-labeled liposomes served as controls (scale bar=100 μm). Histogram shows exosome tissue distribution quantification (n=5/group). CD45, p-Stat3, and CD11b IF staining in liver sections from controls (left) and mice treated with Panc02 EXOs(middle) or Panc02-H7 EXOs (right) for 12 d without tumor challenge. (B) Histogram shows infiltrating CD45 + cell quantification. FN and α-SMA IF staining in liver sections from controls (top) and mice treated with Panc02 EXOs(middle) or Panc02-H7 EXOs (bottom) for 12 d without tumor challenge. (C) Histogram shows infiltrating α-SMA + hStCs and FN expression quantification(400× magnification; n=5/group). Western blotting analysis showed upregulated S100A8 and S100A9 in livers treated with Panc02-H7-derived exosomes. Histogram shows expression of the three proteins in three groupsas determined by densitometric analysis (n=3/group). (D) Pancreatic cancer-derived exosomes induce MDSC accumulation in peripheral blood. (E) Representative flow cytometric plots (left) and quantification (right) of CD11b + GR1 + MDSCs (n=5/group). *P<0.05, **P<0.01,***P<0.001.

Article Snippet: Primary antibodies against fibronectin (1:100),α-SMA (1:50), S100A8 (1:50), S100A9 (1:50), and F4/80 (1:50) were purchased from Proteintech (Wuhan, China).

Techniques: Confocal Microscopy, Labeling, Liposomes, Staining, Expressing, Western Blot, Derivative Assay

Journal: Oncotarget

Article Title: Pancreatic cancer-derived exosomes promote tumor metastasis and liver pre-metastatic niche formation

doi: 10.18632/oncotarget.18831

Figure Lengend Snippet: IHC analysis and histopathological examination of macrophages (F4/80), hStCs (α-SMA), and neutrophils in liver metastatic niches of naïve mice and mice treated with PBS, Panc02 EXOs, or Panc02-H7 EXOs at 30d post-SOI (arrow shows neutrophils in liver) (A) . Representative histogram shows quantification of F4/80 + macrophages, α-SMA + hStCs, and neutrophils (B) . Identification of FN, S100A8, and S100A9 as inflammatory mediators, and collagen deposition in the liver metastatic niche (C) . Representative histogram shows FN and MTS quantification (D) . Representative histogram shows S100A8 and S100A9 quantification (E) . n=6/group.**P<0.01,***P<0.001.10 fields assessed per sample. FOV, field of view.

Article Snippet: Primary antibodies against fibronectin (1:100),α-SMA (1:50), S100A8 (1:50), S100A9 (1:50), and F4/80 (1:50) were purchased from Proteintech (Wuhan, China).

Techniques: