Journal: International Journal of Molecular Sciences

Article Title: A Plant-Derived Antigen–Antibody Complex Induces Anti-Cancer Immune Responses by Forming a Large Quaternary Structure

doi: 10.3390/ijms21165603

Figure Lengend Snippet: Suppression of tumor growth in nude mice through administration of sera from mice immunized with GA P × CO P . BALB/c nu/nu mice were intradermally injected with 1 × 10 6 SW620 cells. Mice were subsequently injected with serum obtained from mice immunized with 1× PBS, GA M , GA P , GA M + CO M , or GA P × CO P (S (1× PBS), S (GA M ), S (GA P ), S (GA M + CO M ), or S (GA P × CO P ), respectively). The positive control was the CO M antibody treated group. At day 1, 4, and 7, all mice were injected with three additional doses of serum or CO M antibody. Tumor volumes (mm 3 ) were recorded at 8, 10, 12, and 15 days after the initial inoculation of cancer cells. The asterisks indicate statistically significant differences (** p < 0.01).

Article Snippet: The SW620 human colorectal carcinoma cells (1 × 10 6 ) were intradermally inoculated into the back of six-week-old BALB/c nu/nu mice (Japan SLC Inc., Hamamatsu, Shizuoka, Japan).

Techniques: Injection, Positive Control

Journal: The American Journal of Pathology

Article Title: KH-Type Splicing Regulatory Protein Controls Colorectal Cancer Cell Growth and Modulates the Tumor Microenvironment

doi: 10.1016/j.ajpath.2019.07.004

Figure Lengend Snippet: KHSRP is involved in regulating cell cycle and mitosis, as well as influencing the protumorigenic extracellular environment in vitro. A: Cell cycle analysis of SW480 cells transfected with siRNAs targeting KHSRP (siKHSRP) or Scramble negative control siRNA; a representative histogram of propidium iodide staining from one experiment is shown, along with a bar graph of cell distribution into cell cycle phases across three replicate experiments. An analysis of variance was performed. B: Analysis of mitotic cells by quantitation of phosphorylated histone H3 in SW480 cells transfected as in A; a representative histogram is shown with bar graph of three replicate experiments. The boxed areas indicate mitotic cells (positive for phospho histone H3 and with 2X DNA content). A Mann-Whitney test was performed. C: Wound-healing assay of SW480 cells transfected as in A; representative images from one experiment are shown, with quantitation of wound area from two replicate experiments. D: Invasion of SW620 cells transfected with siKHSRP or Scramble negative control siRNA, measured by the capacity to migrate through Matrigel; quantification of invaded cells from three replicate experiments is shown, along with representative images. E: Endothelial tube formation of human dermal microendothelial cells (HDECs) treated with conditioned media (CM) from SW620 cells transfected as in D. Tube formation is expressed as percentage of control (HDECs treated with the same media without previous conditioning in SW620 cells). A t-test was performed. F: Expression of secreted IL-8 and vascular endothelial growth factor (VEGF) in the CM of SW620 cells transfected as in D, measured by enzyme-linked immunosorbent assay. A t-test was performed. *P < 0.05, **P < 0.01. Original magnification, ×10 (C–F). AU, arbitrary unit.

Article Snippet: Shotgun Proteomics Stably transfected SW620 cells were treated with vehicle or 500 ng/mL doxycycline for 4 days to induce shRNA-mediated knockdown of KHSRP , as outlined above.

Techniques: In Vitro, Cell Cycle Assay, Transfection, Negative Control, Staining, Quantitation Assay, MANN-WHITNEY, Wound Healing Assay, Control, Expressing, Enzyme-linked Immunosorbent Assay

Journal: The American Journal of Pathology

Article Title: KH-Type Splicing Regulatory Protein Controls Colorectal Cancer Cell Growth and Modulates the Tumor Microenvironment

doi: 10.1016/j.ajpath.2019.07.004

Figure Lengend Snippet: KHSRP regulates expression of key proteins that modulate the tumorigenic microenvironment. Differential expression of secreted proteins was analyzed by shotgun proteomics in conditioned media from SW620 cells treated with doxycycline for 4 days to induce shRNA-mediated KHSRP knockdown. A: Volcano plot showing differentially regulated proteins in the doxycycline-treated samples compared with control, colored in red (log2 fold change > |1|; P < 0.05). Proteins that are differentially regulated above the fold change threshold with a nonsignificant P value are colored in yellow. Dashed lines indicate the thresholds on both axes. B: Dot plot of results from a Gene Ontology (GO) analysis of the 40 highly differentially regulated proteins. GO terms are reported with their category and plotted by gene ratio (the number of genes in one GO term compared with the total). Dots are sized in proportion to the number of hits within a GO term and colored by P value. C: Label-free quantitation of relative abundance of the indicated five proteins in the doxycycline-treated versus control samples. D: Western blot analysis of Ephrin B2 and S100 calcium-binding protein A11 (S100-A11) in conditioned media from control or doxycycline-treated SW620 cells. A Coomassie stain is shown for protein loading control. ****P < 0.0001 (t-test). BP, biological process; CC, cellular component; KP, Kegg pathway; MF, molecular function; RhoGDIα, Ras homolog family–specific guanine nucleotide dissociation inhibitor–α; UP, uniprot annotation.

Article Snippet: Shotgun Proteomics Stably transfected SW620 cells were treated with vehicle or 500 ng/mL doxycycline for 4 days to induce shRNA-mediated knockdown of KHSRP , as outlined above.

Techniques: Expressing, Quantitative Proteomics, shRNA, Knockdown, Control, Quantitation Assay, Western Blot, Binding Assay, Staining

Journal: Oncology Letters

Article Title: Anterior gradient 2 as a supervisory marker for tumor vessel normalization induced by anti-angiogenic treatment

doi: 10.3892/ol.2018.8996

Figure Lengend Snippet: AuNPs delayed tumor growth. (A) Schematic of the study design. SW620-bearing mice were treated daily from day 1–6, and then every other day from day 8–14, with AuNPs (1.3 mg/kg) when the tumor size reached 175–200 mm 3 (designated as day 0 and control mice). Control mice were injected with saline. Tumors were harvested and examined at days 0, 4, 6, 9 and 14 (n=3 per time point). Blood samples were collected from the retro-orbital plexus at days 0, 2, 4, 6, 9, 12 and 14 (n=6 per time point). (B) Quantitative evaluation of body weight and tumor growth after treatment with AuNPs and saline. AuNPs treatment allowed retention of comparable body weights (P>0.05) but delayed tumor growth (*P<0.05). AuNPs, gold nanoparticles.

Article Snippet: SW620 metastatic colorectal cancer cells were purchased from Shanghai Zzbio Co., Ltd. (Shanghai, China).

Techniques: Control, Injection, Saline

Journal: Oncology Letters

Article Title: Anterior gradient 2 as a supervisory marker for tumor vessel normalization induced by anti-angiogenic treatment

doi: 10.3892/ol.2018.8996

Figure Lengend Snippet: Gold nanoparticles (AuNPs) decreased vessel density and reconstructed vessel morphology. (A) CD31 staining of SW620 tumor from control (day 0) and AuNP-treated mice (days 4, 6, 9 and 14). Tumor tissues were stained with an anti-CD31 antibody (red). Scale bars: 50 µm. (B) Quantification of vessel density. Eight random fields were analyzed. Vessel density were measured and presented as CD31-positive areas per field. Relative CD31 levels in tumor at days 6 and 9 vs. controls (*P<0.05). (C) Representative images of SW620 tumor vessels (days 0 and 9). Tumor vessel in saline-treated tumor (day 0) was collapsed (arrows) on the peripheral region but dilated and discontinued (triangles) in the central region. Vessels in AuNPs-treated tumors (day 9) were well organized and structured. Scale bars, 200 µm. AuNPs, gold nanoparticles; CD, cluster of differentiation.

Article Snippet: SW620 metastatic colorectal cancer cells were purchased from Shanghai Zzbio Co., Ltd. (Shanghai, China).

Techniques: Staining, Control, Saline