Journal: Frontiers in Pharmacology

Article Title: Natural Polysaccharides and Their Derivates: A Promising Natural Adjuvant for Tumor Immunotherapy

doi: 10.3389/fphar.2021.621813

Figure Lengend Snippet: The immunoregulatory activity of natural polysaccharides and their derivates on immune cells.

Article Snippet: , C57BL/6 (6 weeks old), BALB/c, OT-I and OT-II TCR transgenic mice and C57BL/6-Ly5.1 (CD45.1) congenic mice; TLR2, TLR4 and SR-A-KO mice; the murine melanoma cell line B16F10 (ATCC, CRL-6475) expressing OVA (B16-OVA) and murine carcinoma cell line CT26 (ATCC, CRL-2639) , Increase levels of co-stimulatory molecule expression and pro-inflammatory cytokine production in spleen DCs dependent on TLR4; enhance ovalbumin (OVA) antigen (Ag)-specific immune activation in tumor-bearing mice , .

Techniques: Activity Assay, Functional Assay, Expressing, Activation Assay, Membrane, Gene Expression, In Vivo, Protein-Protein interactions, Concentration Assay, Transgenic Assay, In Vitro, Phospho-proteomics, Isolation, Cell Culture, Modification, Control, Algae

Journal: Cancer letters

Article Title: Myristoylated Alanine-Rich protein Kinase C Substrate (MARCKS) expression modulates the metastatic phenotype in human and murine colon carcinoma in vitro and in vivo.

doi: 10.1016/j.canlet.2013.01.040

Figure Lengend Snippet: Fig. 5. Down-regulation of MARCKS expression inhibits migration and invasion of CT26 in vitro . (A) Western blot analysis showed the transfection efficiency of CT26 cells stable shRNA-depleted for MARCKS with different shRNA MARCKS (constructs 1–3), shRNA-C is empty vector and shRNA-SC is the scrambled control. (B) A Boyden chamber chemotaxis assay was used with mHSC-CM as chemoattractant. Values are shown as mean ± SD of three independent experiments. P < 0.005 vs shRNA scrambled control. (C) Invasion Matrigel assays were performed using mHSC-CM as chemoattractant. Values are shown as mean ± SD of three independent experi- ments. Invasion was reduced when mHSC-CM was used as chemoattractant. P < 0.005 vs shRNA scrambled control cells.

Article Snippet: Stable gene silencing : Murine colon carcinoma CT26 cells were stable transfected with murine validated short Hairpin RNA (TR512267, 4 shRNA constructs with puromycin cassette, containing two control shRNA) (OriGene Technologies, MD).

Techniques: Expressing, Migration, In Vitro, Western Blot, Transfection, shRNA, Construct, Plasmid Preparation, Control, Chemotaxis Assay

Journal: Cancer letters

Article Title: Myristoylated Alanine-Rich protein Kinase C Substrate (MARCKS) expression modulates the metastatic phenotype in human and murine colon carcinoma in vitro and in vivo.

doi: 10.1016/j.canlet.2013.01.040

Figure Lengend Snippet: Fig. 6. Down-regulation of MARCKS protein delays duration time of mitosis in CT26 cells. (A) Phase contrast imaging shows that shRNA MARCKS-depleted cells obtain a mesenchymal-like phenotype during interphase in contrast to the epithelial-like morphology observed in scrambled control cells. (B) A down-regulation in MARCKS protein expression inhibits cell cycle as was measured by MTT assay (two independent experiments performed each in 6-fold). Values are shown as mean ± SD, P < 0.005 vs scrambled control. (C) A Montage of live cell imaging of shRNA scrambled control cells. Numbers in images indicate time (in minutes) since entering in mitosis. (D) Snapshots of a montage shows that shRNA MARCKS-depleted cells are marked with a delay in DTM that coincides with a morphological aberrant phenotype of the two daughter cells. (E) shRNA MARCKS stable transfected cells show a significant increase in the duration time of mitosis (DTM) when compared to scrambled control cells (sum of DTM, values are shown as mean ± SD, P < 0.005 vs scrambled control). Scrambled control cells have an average DTM of 160 min (marked as t time = 100%) whereas 51.2% of MARCKS depleted cells have a delay in DTM (marked as >t) (n = 3). (F) Representative Western blot analysis shows the absence of MARCKS protein expression in stable transfected shRNA MARCKS-depleted cells (n = at least 3).

Article Snippet: Stable gene silencing : Murine colon carcinoma CT26 cells were stable transfected with murine validated short Hairpin RNA (TR512267, 4 shRNA constructs with puromycin cassette, containing two control shRNA) (OriGene Technologies, MD).

Techniques: Imaging, shRNA, Control, Expressing, MTT Assay, Live Cell Imaging, Transfection, Western Blot

Journal: Cancer letters

Article Title: Myristoylated Alanine-Rich protein Kinase C Substrate (MARCKS) expression modulates the metastatic phenotype in human and murine colon carcinoma in vitro and in vivo.

doi: 10.1016/j.canlet.2013.01.040

Figure Lengend Snippet: Fig. 7. The effect of MARCKS-depletion in CT26 cells attenuates tumour progression in vivo . (A) Representative Western blot analysis shows the stable transfection efficiency of shRNA against MARCKS in CT26 cells. (B) Photographs of representative liver after performing a colorectal liver metastasis assay in vivo . In both conditions, tumour formation was observed in the intrasplenic injection site, whereas hepatic metastases were detectable in SCRNA CT26 tumour-challenged mice with few or no tumour formation in MARCKS-depleted CT26 cells tumour-challenged mice. (C) Graphs show the body–liver weight (%) and demonstrates a significant induction of CT26 and MARCKS-depleted CT26 tumour-challenged mice vs sham-operated (P < 0.05, vs sham-operated mice). Tumour-challenged mice with shRNA MARCKS-depleted CT26 cells showed few or no metastases in comparison to CT26 tumour-challenged mice (P < 0.005 vs shRNA SC). (D) Haematoxylin-eosin staining confirmed the presence of hepatic tumours in CT26 tumour-challenged mice.

Article Snippet: Stable gene silencing : Murine colon carcinoma CT26 cells were stable transfected with murine validated short Hairpin RNA (TR512267, 4 shRNA constructs with puromycin cassette, containing two control shRNA) (OriGene Technologies, MD).

Techniques: In Vivo, Western Blot, Stable Transfection, shRNA, Injection, Comparison, Staining

Journal: Cancer Research and Treatment : Official Journal of Korean Cancer Association

Article Title: The Oncogenic Role of TNFRSF12A in Colorectal Cancer and Pan-Cancer Bioinformatics Analysis

doi: 10.4143/crt.2024.408

Figure Lengend Snippet: Tumor necrosis factor receptor superfamily member 12A (TNFRSF12A) effects on growth, colony formation, and migration of colorectal cancer cell lines. (A) Quantitative real-time polymerase chain reaction to assess the mRNA levels of TNFRSF12A in the inducible TNFRSF12A knockdown or overexpression colorectal cancer cell lines. The duration of doxycycline (Dox) treatment was 48 hours. (B) Western blot to assess the protein levels of TNFRSF12A in the inducible TNFRSF12A knockdown or overexpression colorectal cancer cell lines. The duration of Dox treatment was 48 hours. (C) Growth curves of colorectal cancer cell lines with TNFRSF12A knockdown or overexpression. (D) Colony formation results of colorectal cancer cell lines with TNFRSF12A knockdown or overexpression. Scale bars=5 mm. (E) Wound healing results of colorectal cancer cell lines with TNFRSF12A knockdown or overexpression. Scale bars=250 μm. The notation “sh” refers to TNFRSF12A shRNA, and “oeT” indicates overexpressed TNFRSF12A. Statistical significance levels are denoted as follows: * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; ns, not significant.

Article Snippet: Colorectal cancer cell lines (human: DLD-1, RKO, and HT-29; mouse: CT26) and HEK293T were obtained from the American Type Culture Collection (ATCC).

Techniques: Migration, Real-time Polymerase Chain Reaction, Knockdown, Over Expression, Western Blot, shRNA

Journal: Cancer Research and Treatment : Official Journal of Korean Cancer Association

Article Title: The Oncogenic Role of TNFRSF12A in Colorectal Cancer and Pan-Cancer Bioinformatics Analysis

doi: 10.4143/crt.2024.408

Figure Lengend Snippet: Tumor necrosis factor receptor superfamily member 12A (TNFRSF12A) promotes colorectal cancer growth in vivo . (A) Quantitative real-time polymerase chain reaction to assess the mRNA levels of TNFRSF12A in the inducible TNFRSF12A knockdown or overexpression CT26 mouse colorectal cancer cells. The duration of Dox treatment was 48 hours. (B) Western blot to assess the protein levels of TNFRSF12A in the inducible TNFRSF12A knockdown or overexpression CT26 mouse colorectal cancer cells. The duration of Dox treatment was 48 hours. (C) Growth curves of CT26 cells with TNFRSF12A knockdown or overexpression. (D) Representative colony formation images of CT26 cells with TNFRSF12A knockdown or overexpression. Scale bars=5 mm. (E) Representative wound healing images of CT26 cells with TNFRSF12A knockdown or overexpression. Scale bars=250 μm. (F) Images of tumors obtained from mice that were subcutaneously transplanted with CT26 cells with either TNFRSF12A knockdown or overexpression (n=5). (G) Growth curves of tumors in mice subcutaneously transplanted with CT26 cells with either TNFRSF12A knockdown or overexpression. (H) The weight of tumors obtained from mice that were subcutaneously transplanted with CT26 cells with either TNFRSF12A knockdown or overexpression. The notation “sh#1” refers to TNFRSF12A shRNA#1, and “oeT” indicates overexpressed TNFRSF12A. Statistical significance levels are denoted as follows: * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; ns, not significant.

Article Snippet: Colorectal cancer cell lines (human: DLD-1, RKO, and HT-29; mouse: CT26) and HEK293T were obtained from the American Type Culture Collection (ATCC).

Techniques: In Vivo, Real-time Polymerase Chain Reaction, Knockdown, Over Expression, Western Blot, shRNA

Journal: Cancer Research and Treatment : Official Journal of Korean Cancer Association

Article Title: The Oncogenic Role of TNFRSF12A in Colorectal Cancer and Pan-Cancer Bioinformatics Analysis

doi: 10.4143/crt.2024.408

Figure Lengend Snippet: Differential expression genes and enrichment analysis of colorectal cancer cell line with or without tumor necrosis factor receptor superfamily member 12A (TNFRSF12A) overexpression. (A) The heatmap of differential expression genes between TNFRSF12A overexpressed (oeT) DLD-1 cells and the control (oeCtl) DLD-1 cells. (B) The volcano plot of differential expression genes between TNFRSF12A overexpressed (oeT) DLD-1 cells and the control (oeCtl) DLD-1 cells. (C) The results gene ontology (GO) analysis for differential expression genes between TNFRSF12A overexpressed (oeT) DLD-1 cells and the control (oeCtl) DLD-1 cells. BP, biological process; MF, molecular function. (D) Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis for differential expression genes between TNFRSF12A overexpressed (oeT) DLD-1 cells and the control (oeCtl) DLD-1 cells. (E) The heatmap of the expression of nuclear factor κB (NF-κB)–related genes in TNFRSF12A overexpressed (oeT) DLD-1 cells and the control (oeCtl) DLD-1 cells. (F) The heatmap of the expression of putative NF-κB targets in TNFRSF12A overexpressed (oeT) DLD-1 cells and the control (oeCtl) DLD-1 cells.

Article Snippet: Colorectal cancer cell lines (human: DLD-1, RKO, and HT-29; mouse: CT26) and HEK293T were obtained from the American Type Culture Collection (ATCC).

Techniques: Quantitative Proteomics, Over Expression, Control, Expressing

Journal: Cancer Research and Treatment : Official Journal of Korean Cancer Association

Article Title: The Oncogenic Role of TNFRSF12A in Colorectal Cancer and Pan-Cancer Bioinformatics Analysis

doi: 10.4143/crt.2024.408

Figure Lengend Snippet: Baculoviral IAP repeat containing 3 (BIRC3) is upregulated after tumor necrosis factor receptor superfamily member 12A (TNFRSF12A) overexpression and a potential target of RELA. (A) Quantitative real-time polymerase chain reaction verification of BIRC3 expression in DLD-1 and RKO cells with TNFRSF12A knockdown and overexpression. The doxycycline (Dox) treatment duration was 48 hours. The notation “sh#1” refers to TNFRSF12A shRNA#1, and “oeT” indicates overexpressed TNFRSF12A. Statistical significance levels are denoted as follows: ** p < 0.01, *** p < 0.001, **** p < 0.0001. (B) Profile of RELA binding peaks on BIRC3 gene in LoVo colorectal cancer cells from ChIP-Atlas database (SRX359916). (C) Potential relationships among TNFRSF12A, BIRC3, and RELA in STRING database.

Article Snippet: Colorectal cancer cell lines (human: DLD-1, RKO, and HT-29; mouse: CT26) and HEK293T were obtained from the American Type Culture Collection (ATCC).

Techniques: Over Expression, Real-time Polymerase Chain Reaction, Expressing, Knockdown, shRNA, Binding Assay

Journal: Cancer Research and Treatment : Official Journal of Korean Cancer Association

Article Title: The Oncogenic Role of TNFRSF12A in Colorectal Cancer and Pan-Cancer Bioinformatics Analysis

doi: 10.4143/crt.2024.408

Figure Lengend Snippet: Tumor necrosis factor receptor superfamily member 12A (TNFRSF12A) overexpression affects colorectal cancer cell growth, colony formation, and migration through TNFRSF12A/RELA/BIRC3 axis. (A) Quantitative real-time polymerase chain reaction verification of the efficiency of transient knockdown BIRC3 in DLD-1 and RKO cells. The doxycycline (Dox) treatment duration was 48 hours. (B) The growth curves of TNFRSF12A overexpression DLD-1 and RKO cells with transient BIRC3 knockdown. (C) Representative colony formation images of TNFRSF12A overexpression DLD-1 and RKO cells with transient BIRC3 knockdown. Scale bars=5 mm. (D) Representative wound healing assay images of TNFRSF12A overexpression DLD-1 and RKO cells with transient BIRC3 knockdown. Scale bars=250 μm. (E) Western blot to detect level of TNFRSF12A and BIRC3, p-RELA and RELA in tumor from TNFRSF12A overexpression group, homograft mouse model mentioned in . (F) Western blot to detect levels of TNFRSF12A and BIRC3, p-RELA and RELA in DLD-1 and RKO cell lines with or without TNFRSF12A overexpression and RELA inhibitor JSH-23. The treatment duration was 48 hours. The notation “oeT” indicates overexpressed TNFRSF12A. Statistical significance levels are denoted as follows: * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; ns, not significant.

Article Snippet: Colorectal cancer cell lines (human: DLD-1, RKO, and HT-29; mouse: CT26) and HEK293T were obtained from the American Type Culture Collection (ATCC).

Techniques: Over Expression, Migration, Real-time Polymerase Chain Reaction, Knockdown, Wound Healing Assay, Western Blot

Journal: Cancer Research and Treatment : Official Journal of Korean Cancer Association

Article Title: The Oncogenic Role of TNFRSF12A in Colorectal Cancer and Pan-Cancer Bioinformatics Analysis

doi: 10.4143/crt.2024.408

Figure Lengend Snippet: Scheme of inferred tumor necrosis factor receptor superfamily member 12A (TNFRSF12A) function in colorectal cancer. BIRC3, baculoviral inhibitor of apoptosis protein repeat containing 3.

Article Snippet: Colorectal cancer cell lines (human: DLD-1, RKO, and HT-29; mouse: CT26) and HEK293T were obtained from the American Type Culture Collection (ATCC).

Techniques: