Journal: American Journal of Human Genetics

Article Title: Systematic Functional Interrogation of Genes in GWAS Loci Identified ATF1 as a Key Driver in Colorectal Cancer Modulated by a Promoter-Enhancer Interaction

doi: 10.1016/j.ajhg.2019.05.004

Figure Lengend Snippet: ATF1 Promotes Cell Proliferation and Xenograft Tumor Growth in CRC by Affecting Cell Apoptosis

Article Snippet: A modified ATF1 knockout CRC HCT116 cell line was generated by CRISPR-Cas9 technology (Genloci Biotechnologies).

Techniques:

Journal: American Journal of Human Genetics

Article Title: Systematic Functional Interrogation of Genes in GWAS Loci Identified ATF1 as a Key Driver in Colorectal Cancer Modulated by a Promoter-Enhancer Interaction

doi: 10.1016/j.ajhg.2019.05.004

Figure Lengend Snippet: Functional Genomic Screening Reveals that ATF1 Is an Oncogene in CRC

Article Snippet: A modified ATF1 knockout CRC HCT116 cell line was generated by CRISPR-Cas9 technology (Genloci Biotechnologies).

Techniques: Functional Assay

Journal: American Journal of Human Genetics

Article Title: Systematic Functional Interrogation of Genes in GWAS Loci Identified ATF1 as a Key Driver in Colorectal Cancer Modulated by a Promoter-Enhancer Interaction

doi: 10.1016/j.ajhg.2019.05.004

Figure Lengend Snippet: SP1 and GATA3 preferentially bind to the dbSNP: rs61926301[T] and dbSNP: rs7959129[T] alleles at the ATF1 promoter and first intron region, respectively

Article Snippet: A modified ATF1 knockout CRC HCT116 cell line was generated by CRISPR-Cas9 technology (Genloci Biotechnologies).

Techniques:

Journal: American Journal of Human Genetics

Article Title: Systematic Functional Interrogation of Genes in GWAS Loci Identified ATF1 as a Key Driver in Colorectal Cancer Modulated by a Promoter-Enhancer Interaction

doi: 10.1016/j.ajhg.2019.05.004

Figure Lengend Snippet: Transcription Factors SP1 and GATA3 Correlate with ATF1 Expression in an Allele-Specific Manner

Article Snippet: A modified ATF1 knockout CRC HCT116 cell line was generated by CRISPR-Cas9 technology (Genloci Biotechnologies).

Techniques: Expressing

Journal: American Journal of Human Genetics

Article Title: Systematic Functional Interrogation of Genes in GWAS Loci Identified ATF1 as a Key Driver in Colorectal Cancer Modulated by a Promoter-Enhancer Interaction

doi: 10.1016/j.ajhg.2019.05.004

Figure Lengend Snippet: rs61926301 and rs7959129 Synergistically Facilitate ATF1 Transcriptional Activity Mediated by the Transcription Factors SP1 and GATA3

Article Snippet: A modified ATF1 knockout CRC HCT116 cell line was generated by CRISPR-Cas9 technology (Genloci Biotechnologies).

Techniques: Activity Assay

Journal: American Journal of Human Genetics

Article Title: Systematic Functional Interrogation of Genes in GWAS Loci Identified ATF1 as a Key Driver in Colorectal Cancer Modulated by a Promoter-Enhancer Interaction

doi: 10.1016/j.ajhg.2019.05.004

Figure Lengend Snippet: Risk Alleles of rs61926301 and rs7959129 Facilitate a Promoter-Enhancer Interaction Mediated by SP1 and GATA3 to Upregulate ATF1 Expression

Article Snippet: A modified ATF1 knockout CRC HCT116 cell line was generated by CRISPR-Cas9 technology (Genloci Biotechnologies).

Techniques: Expressing

Journal: American Journal of Human Genetics

Article Title: Systematic Functional Interrogation of Genes in GWAS Loci Identified ATF1 as a Key Driver in Colorectal Cancer Modulated by a Promoter-Enhancer Interaction

doi: 10.1016/j.ajhg.2019.05.004

Figure Lengend Snippet: ATF1 Activates a Panel of Genes Associated with Apoptosis, Wnt, TGF-β, and MAPK Pathways and Facilitates the Early Onset of CRC

Article Snippet: A modified ATF1 knockout CRC HCT116 cell line was generated by CRISPR-Cas9 technology (Genloci Biotechnologies).

Techniques:

Journal: American Journal of Human Genetics

Article Title: Systematic Functional Interrogation of Genes in GWAS Loci Identified ATF1 as a Key Driver in Colorectal Cancer Modulated by a Promoter-Enhancer Interaction

doi: 10.1016/j.ajhg.2019.05.004

Figure Lengend Snippet: Graphical Representation of the Regulation and Function of ATF1 in CRC

Article Snippet: A modified ATF1 knockout CRC HCT116 cell line was generated by CRISPR-Cas9 technology (Genloci Biotechnologies).

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Epigenetic Silencing of miR-218-5p Modulates BIRC5 and DDX21 Expression to Promote Colorectal Cancer Progression

doi: 10.3390/ijms26094146

Figure Lengend Snippet: Validation of selected miR-218-5p Gene Targets. ( A ) Validation of BIRC5 and DDX21 as bona fide gene targets for miR-218-5p in CRC. Two-tailed t -test was used to compare different groups. ** p < 0.005, *** p < 0.0005, **** p < 0.00005. ( B ) Western blot showing DDX21 protein expression in miR-218-5p overexpressing compared to control HT-29 and HCT116 cells. Quantification of DDX21 protein expression normalized to ACTB is shown in the right panel. ( C ) Gene effect score based on CRISPR-Cas9 screen data in 40 CRC cell models from the DepMap database.

Article Snippet: A notable observation within SLIT2 , which encodes miR-218-1 within its intronic region, is the detection of higher levels of hypermethylation near the end of the promoter regions in the CRC cell lines, just upstream of the transcription start site, where the TATA box is typically located.

Techniques: Biomarker Discovery, Two Tailed Test, Western Blot, Expressing, Control, CRISPR

Journal: International Journal of Molecular Sciences

Article Title: Epigenetic Silencing of miR-218-5p Modulates BIRC5 and DDX21 Expression to Promote Colorectal Cancer Progression

doi: 10.3390/ijms26094146

Figure Lengend Snippet: Suppression of SLIT2 and SLIT3 miR-218 Host Genes in CRC. ( A ) Genomic location of miR-218-1 and miR-218-2 within the SLIT2 and SLIT3 genomic region on chromosome 4 and 5, respectively. ( B ) Correlation plot between miR-218-5p and SLIT2 ( left ) and SLIT3 ( right ) in a large cohort of COAD (n = 450) from the ENCORI project. ( C ) Downregulation of SLIT2 ( left ) and SLIT3 ( right ) in COAD (n = 275) compared to normal colon tissue (n = 349) from GEPIA2 database. T: tumor, N: normal. * p < 0.05. ( D ) Methylation analysis of SLIT2 and SLIT3 promoters using bisulfite conversion and NGS in a panel of CRC cell models (HCT116, HT-29, SW-480, LoVo, and DLD-1) compared to the MCF10A normal epithelial cells. ( E ) Schematic representation illustrating (🠯) downregulation of SLIT2 and SLIT3 in CRC to lead to miR-218-5p suppression (🠯), thus promoting tumorigenesis due to lifted suppression (🠭) of BIRC5, DDX21, and other gene targets identified in the current study.

Article Snippet: A notable observation within SLIT2 , which encodes miR-218-1 within its intronic region, is the detection of higher levels of hypermethylation near the end of the promoter regions in the CRC cell lines, just upstream of the transcription start site, where the TATA box is typically located.

Techniques: Methylation

Journal: Journal of Translational Medicine

Article Title: Disruption of the c-Myc/miR-200b-3p/PRDX2 regulatory loop enhances tumor metastasis and chemotherapeutic resistance in colorectal cancer

doi: 10.1186/s12967-017-1357-7

Figure Lengend Snippet: MiR-200b-3p inhibits CRC cells’ metastatic capacity by targeting PRDX2 in vivo. a Intestinal and hepatic metastatic nodules after subcutaneous tumors derived from LoVo/NC, LoVo/miR and LoVo/miR + PRDX2 cells were transplanted in the mesentery at the distal end of cecum in mice (n = 5) for 6 weeks. Red arrows point at potential metastatic nodules in intestines. Scale bars represent 50 μm. b , c The number of hepatic metastatic nodules ( b ) or intestinal metastatic nodules ( c ) of mice with tumors derived from LoVo/NC, LoVo/miR and LoVo/miR + PRDX2 cells. The number of hepatic metastatic nodules per mouse was counted under the microscope, with five high power fields (HPF) observation (*** p < 0.001). d Intestinal and hepatic metastatic nodules after subcutaneous tumors derived from SW480/NC and SW480/Zip-miR cells were transplanted in the mesentery at the distal end of cecum in mice (n = 5) for 6 weeks. Red arrows point at potential metastatic nodules in intestines. Scale bars represent 50 μm. e , f The number of hepatic metastatic nodules ( e ) or intestinal metastatic nodules ( f ) of mice with tumors derived from SW480/NC and SW480/Zip-miR cells. The number of hepatic metastatic nodules per mouse was counted under the microscope, with five HPF observation (*** p < 0.001)

Article Snippet: First, 1 × 10 6 CRC cells with different treatments were subcutaneously injected into the female BALB/c nude mice (Beijing HFK Bioscience Co., LTD, Beijing China) aged 5 weeks for 4 weeks to generate subcutaneous tumors and then the mice were sacrificed.

Techniques: In Vivo, Derivative Assay, Microscopy

Journal: Journal of Translational Medicine

Article Title: Disruption of the c-Myc/miR-200b-3p/PRDX2 regulatory loop enhances tumor metastasis and chemotherapeutic resistance in colorectal cancer

doi: 10.1186/s12967-017-1357-7

Figure Lengend Snippet: Disruption of the c-Myc/miR-200b-3p/PRDX2 regulatory loop enhances chemotherapeutic resistance of CRC cells. a c-Myc overexpression enhanced the resistance of SW480 cells to oxaliplatin, while the enhanced effect was dampened by miR-200b-3p partly. b miR-200b-3p overexpression reduced the resistance of LoVo cells to oxaliplatin, while the reduced effect was reversed by nontargetable PRDX2 partly. c c-Myc overexpression repressed oxaliplatin-induced apoptosis in SW480 cells, while the repressive effect was reversed by miR-200b-3p partly. d miR-200b-3p overexpression enhanced oxaliplatin-induced apoptosis in LoVo cells, while the enhanced effect was abolished by nontargetale PRDX2 partly. e , f The oxaliplatin-induced apoptosis was statistically analyzed in LoVo cells ( e ) or in SW480 cells ( f ) among different treatment groups

Article Snippet: First, 1 × 10 6 CRC cells with different treatments were subcutaneously injected into the female BALB/c nude mice (Beijing HFK Bioscience Co., LTD, Beijing China) aged 5 weeks for 4 weeks to generate subcutaneous tumors and then the mice were sacrificed.

Techniques: Disruption, Over Expression

Journal: Neoplasia (New York, N.Y.)

Article Title: Biologic Response of Colorectal Cancer Xenograft Tumors to Sequential Treatment with Panitumumab and Bevacizumab

doi: 10.1016/j.neo.2018.04.006

Figure Lengend Snippet: Antitumor effect in LIM1215(A) xenografts by treatment received. (A and B) assessed by average tumor volume and (C) relative growth rate. Arrowheads indicate dosing. Arrows indicate sample harvesting. Data represent mean ± SE. ( n = 3–7. * P < .05). B, bevacizumab; P, panitumumab; V, vehicle control.

Article Snippet: The human colon cancer cell line LIM1215 was obtained from the European Collection of Authenticated Cell Cultures (ECACC, Salisbury, UK).

Techniques: Control

Journal: Neoplasia (New York, N.Y.)

Article Title: Biologic Response of Colorectal Cancer Xenograft Tumors to Sequential Treatment with Panitumumab and Bevacizumab

doi: 10.1016/j.neo.2018.04.006

Figure Lengend Snippet: Outcome of immunohistochemistry for Ki-67 in LIM1215(B) xenograft sections. (A) Using vehicle control, (B) panitumumab-bevacizumab, (C) bevacizumab-panitumumab, and (D) bevacizumab-bevacizumab. (E) Proportion of Ki-67-positive cells in all treatment groups. Sections were IHC stained for Ki-67 (brown) and counterstained with hematoxylin (purple). Representative images of the sections are shown. Data in the graph represent the mean ± SE ( n = 6–8). ** P < .01. B, bevacizumab; P, panitumumab; V, vehicle control.

Article Snippet: The human colon cancer cell line LIM1215 was obtained from the European Collection of Authenticated Cell Cultures (ECACC, Salisbury, UK).

Techniques: Immunohistochemistry, Control, Staining

Journal: Neoplasia (New York, N.Y.)

Article Title: Biologic Response of Colorectal Cancer Xenograft Tumors to Sequential Treatment with Panitumumab and Bevacizumab

doi: 10.1016/j.neo.2018.04.006

Figure Lengend Snippet: Levels of Phosphorylated Growth Factor Receptors in LIM1215(A) Xenografts Treated with PB, BP, and BB Relative to Vehicle Control

Article Snippet: The human colon cancer cell line LIM1215 was obtained from the European Collection of Authenticated Cell Cultures (ECACC, Salisbury, UK).

Techniques: Phospho-proteomics, Control

Journal: Neoplasia (New York, N.Y.)

Article Title: Biologic Response of Colorectal Cancer Xenograft Tumors to Sequential Treatment with Panitumumab and Bevacizumab

doi: 10.1016/j.neo.2018.04.006

Figure Lengend Snippet: Results of western blotting in LIM1215(B) xenografts. (A) EPHA2 and pEPHA2 with panitumumab-bevacizumab compared with bevacizumab-panitumumab. (B) EPHA2 and pEPHA2 with panitumumab-bevacizumab compared with bevacizumab-bevacizumab. (C) Phosphorylation of EPHA2 for panitumumab-bevacizumab compared with bevacizumab-panitumumab and (D) compared with bevacizumab-bevacizumab. (E) RSK and pRSK with panitumumab-bevacizumab compared with bevacizumab-panitumumab and (F) RSK and pRSK with panitumumab-bevacizumab compared with bevacizumab-bevacizumab. (G) Phosphorylation of RSK for panitumumab-bevacizumab compared with bevacizumab-panitumumab and (H) compared with bevacizumab-bevacizumab. Data represent mean ± SD ( n = 8). ** P < .01, *** P < .001. B, bevacizumab; P, panitumumab; V, vehicle control.

Article Snippet: The human colon cancer cell line LIM1215 was obtained from the European Collection of Authenticated Cell Cultures (ECACC, Salisbury, UK).

Techniques: Western Blot, Phospho-proteomics, Control

Journal: Neoplasia (New York, N.Y.)

Article Title: Biologic Response of Colorectal Cancer Xenograft Tumors to Sequential Treatment with Panitumumab and Bevacizumab

doi: 10.1016/j.neo.2018.04.006

Figure Lengend Snippet: Enrichment Analysis of LIM1215(A) Xenografts Treated with Bevacizumab-Bevacizumab Compared with Vehicle Control (all Canonical Pathways, P < .001)

Article Snippet: The human colon cancer cell line LIM1215 was obtained from the European Collection of Authenticated Cell Cultures (ECACC, Salisbury, UK).

Techniques: Control, Activation Assay, Inhibition

Journal: Neoplasia (New York, N.Y.)

Article Title: Biologic Response of Colorectal Cancer Xenograft Tumors to Sequential Treatment with Panitumumab and Bevacizumab

doi: 10.1016/j.neo.2018.04.006

Figure Lengend Snippet: Relative expression of (A–H) lipogenic ( FASN, HMGCR, MVD, LSS ) and (I–L) hypoxia-related ( CA9, TGFBI ) genes in LIM1215(B) xenograft tumors. Expression relative to vehicle control with first-line treatment is shown in (A–D) and (I–J), and with sequential treatment in (E–H) and (K–L). Data represent mean ± SD ( n = 8). * P < .05, ** P < .01. B, bevacizumab; P, panitumumab; V, vehicle control.

Article Snippet: The human colon cancer cell line LIM1215 was obtained from the European Collection of Authenticated Cell Cultures (ECACC, Salisbury, UK).

Techniques: Expressing, Control

Journal: Biomedicines

Article Title: Acetate-Mediated Odorant Receptor OR51E2 Activation Results in Calcitonin Secretion in Parafollicular C-Cells: A Novel Diagnostic Target of Human Medullary Thyroid Cancer

doi: 10.3390/biomedicines11061688

Figure Lengend Snippet: C-cell-specific OR51E2 expression increases in medullary thyroid cancer. ( A ) Immunofluorescence staining of calcitonin (green) and olfactory marker protein (OMP, red) in normal human thyroid and medullary thyroid cancer tissue (scale bar: 10 µm). ( B ) mRNA expression levels of OR51E2, OMP, calcitonin, and OR51E1 were measured in medullary and anaplastic/papillary/follicular cancer cell lines. GAPDH was used to normalize expression, and relative expression was confirmed using the 2 −ΔΔCt method based on FRO values. Data are presented as relative density (fold) compared to FRO cell lines. One-way ANOVA, n ≥ 3, (** p < 0.01; and *** p < 0.005; **** p < 0.001). ( C ) Immunofluorescent staining demonstrated co-localization of OR51E2 (green) and OMP (red) in the MZ-CRC-1 and TT (medullary thyroid cancer [MTC] cell lines) (scale bar: 10 µm). ( D , E ) Immunohistochemical staining ( D ) and Western blotting analysis ( E ) revealed that OR51E2 was expressed in normal thyroid and MTC tissue in a 31-year-old patient with MTC (scale bar: 10 μm).

Article Snippet: Briefly, 200 μL of MZ-CRC-1 cells (1 × 10 4 cells/well) was seeded in white polystyrene 96-well plates (#353296; BD Biosciences, San Jose, CA, USA) 1 day before transfection.

Techniques: Expressing, Immunofluorescence, Staining, Marker, Immunohistochemical staining, Western Blot

Journal: Biomedicines

Article Title: Acetate-Mediated Odorant Receptor OR51E2 Activation Results in Calcitonin Secretion in Parafollicular C-Cells: A Novel Diagnostic Target of Human Medullary Thyroid Cancer

doi: 10.3390/biomedicines11061688

Figure Lengend Snippet: Acetate-stimulated OR51E2 increases calcitonin secretion through the cAMP pathway. ( A ) Relative luciferase activities of OR51E2 vs. concentrations of the indicated acetate in MZ-CRC-1 cells. Data are expressed as mean ± standard error of mean (SEM) ( n = 5) at each concentration. ( B – D ) Acetate-OR51E2 induced calcitonin secretion through the cAMP pathway. ( B ) cAMP levels were measured in MZ-CRC-1 cells 1 h after 250 µM acetate treatment. All data are expressed as mean ± SEM. ****, p < 0.0001 by one-way ANOVA with Student’s t -test. ( C ) Western blot with anti-phospho-CREB antibody showing the phosphorylation of CREB in the MZ-CRC-1 cells 15 min after acetate treatment. β-actin was used as the loading control. ( D ) OR51E2 siRNA or non-targeting siRNA (siCtrl) was transfected into MZ-CRC-1 cells for 48 h. Medium was collected in a time-dependent manner (0, 1, 2, 3, 4, and 5 min) after 250 µM acetate treatment, and the calcitonin levels were measured using an ELISA kit. RT-PCR analysis of OR51E2 mRNA expression in MZ-CRC-1 cells transfected with siCtrl or siOR51E2 (bottom).

Article Snippet: Briefly, 200 μL of MZ-CRC-1 cells (1 × 10 4 cells/well) was seeded in white polystyrene 96-well plates (#353296; BD Biosciences, San Jose, CA, USA) 1 day before transfection.

Techniques: Luciferase, Concentration Assay, Western Blot, Transfection, Enzyme-linked Immunosorbent Assay, Reverse Transcription Polymerase Chain Reaction, Expressing

Journal: Biomedicines

Article Title: Acetate-Mediated Odorant Receptor OR51E2 Activation Results in Calcitonin Secretion in Parafollicular C-Cells: A Novel Diagnostic Target of Human Medullary Thyroid Cancer

doi: 10.3390/biomedicines11061688

Figure Lengend Snippet: Acetate is internalized with OR51E2 in the MTC. ( A ) Nuclear translocation of OR51E2 after acetate treatment in MZ-CRC-1 cells—serum-starved MZ-CRC-1 cells were incubated with acetate at indicated times, and cells were immunostained with anti-OR51E2 antibody (scale bar: 10 μm). ( B ) C 11 -acetate uptake assay in thyroid cancer cell lines revealed the highest uptake in MZ-CRC-1 cells. The cells were treated with C 11 -acetate (at 5 µCi) for 10 min; they were collected and counted using a gamma counter. The results were normalized with protein concentration. Mann–Whitney test, p < 0.001 for MZ-CRC-1 vs. all the other cell lines, p < 0.001 for TT vs. TPC-1, and p = 0.038 for TT vs. FTC-133 ( n = 3). ( C ) Dose-dependent C 11 -acetate uptake in the medullary thyroid cancer cell lines (top panels). The cells were treated with C 11 -acetate (2, 5, 10, 20, 40, and 60 µCi) for 10 min. For the C 11 -acetate competitive binding assay (bottom panels), non-radioactive acetate treatment was performed for 3 min in a dose-dependent manner. After PBS washing, 10 µCi of C 11 -acetate treatment for 10 min was performed. Cells were collected, counted using a gamma counter, and normalized using the protein concentration. ( D ) In vitro assay (up) C 11 -acetate uptake in OR51E2 siRNA knockdown MZ-CRC-1 cells decreases significantly. After transfection of MZ-CRC-1 cells with non-targeting siRNA (siCtrl) or siOR51E2 for 72 h, cells were incubated with 10 µCi C 11 -acetate for 10 min, and the C 11 -acetate uptake was measured. Cells were harvested, counted using a gamma counter, and counts in the samples were normalized with intracellular protein concentration. All data are means ± SEM ( n = 3). * p < 0.05 using one-way ANOVA with a paired t -test. In vivo radiotracer assay (bottom) C 11 -acetate uptake decreased in Olfr78 -KO mice compared to WT-tumor-bearing mice ( n = 4). C 11 -acetate uptake was confirmed in eight-week-old Olfr78 -KO and WT mice. C 11 -acetate was injected into the tail vein, and the mice were sacrificed 10 min later. The C 11 -acetate amounts in thyroid gland and orbital blood were counted using a gamma counter and normalized using blood values. All the data are means ± SEM ( n = 3). * p < 0.05 using one-way ANOVA and the Mann–Whitney test.

Article Snippet: Briefly, 200 μL of MZ-CRC-1 cells (1 × 10 4 cells/well) was seeded in white polystyrene 96-well plates (#353296; BD Biosciences, San Jose, CA, USA) 1 day before transfection.

Techniques: Translocation Assay, Incubation, Protein Concentration, MANN-WHITNEY, Competitive Binding Assay, In Vitro, Transfection, In Vivo, Injection

Journal: Cell Reports Medicine

Article Title: Combined therapy with DR5-targeting antibody-drug conjugate and CDK inhibitors as a strategy for advanced colorectal cancer

doi: 10.1016/j.xcrm.2025.102158

Figure Lengend Snippet: Combination effect of Oba01 and Abem on MSS and MSI-H CRC cell lines in vitro (A) Viability of cells treated with Oba01 (μg/mL) either alone or in combination (COM) with Abem (μM). (B–D) Survival of cells treated with indicated drugs determined by confluency using the Incucyte S3 system (B), with confluency (%) at 72 h shown in (C) and representative images in (D). Scale bars, 900 μm. (E and F) Flow cytometry analysis of apoptosis, with Annexin V/PI-positive cells shown in bar charts. (G) KEGG analysis of DEGs in the COM-treated group vs. vehicle in P53R cells. (H) GSEA analysis showing enrichment of cell death-related pathways in P53R cells under combination treatment. (I) Expression of key molecules in the anti-apoptotic regulation, cell cycle, and MTOR signaling pathways. Data represent the mean ± SEM of ≥3 experiments. p values from one-way ANOVA. ns, non-significant. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001.

Article Snippet: The human CRC cells HCT116, P53R, RKO, COLO205 and HEK293T were purchased from iCell Bioscience Inc. (Shanghai, China).

Techniques: In Vitro, Flow Cytometry, Expressing, Protein-Protein interactions

Journal: Gut

Article Title: Circulating microRNA-203 predicts prognosis and metastasis in human colorectal cancer

doi: 10.1136/gutjnl-2014-308737

Figure Lengend Snippet: Athymic nude mice for liver metastatic models were sacrificed 10 weeks after injection of HT-29 CRC cells (2.0 × 106/50 μL of PBS) in the spleen. (A) Representative images of non-treated control mouse. (B) Representative images of animals with established liver metastasis. Yellow arrows indicate metastatic foci. The upper-right panel illustrates representative results of H&E staining in mouse liver. The lower-left panel displays representative results for ISH staining for miR-203 in animal liver. (C) miR-203 expression status in spleen (S), adjacent hepatocytes (H), and liver metastasis (LM) tissues of liver metastasis established mouse group (n=10; *** P<0.001). (D) Serum miR-203 expression status between non-treated control animals (non-LM; n=7) and animals with established liver metastasis (LM; n=10; * P<0.05). Athymic nude mice for systemic metastatic models were sacrificed 8 weeks after injection of HT-29 CRC cells (1.0 × 106/200 μL of PBS) in the tail vein. Representative images of non-treated controls (E) and established systemic metastatic mouse (F). Red arrows indicate metastatic foci. (G) Serum miR-203 expression status between non-treated control animals (non-SM; n=6) and animals with established systemic metastasis (SM; n=6; ** P<0.01). (H) Significant correlation between serum miR-203 expression and tumor volume in mice (Rho = 0.829, P=0.0009).

Article Snippet: Additionally, CRC cells (2.0 × 10 6 /50 μL of PBS) were injected in the spleen of six-week-old male athymic nude mice (Balb/c nu; Harlan Laboratories, Inc.) during an open laparotomy to establish in vivo mice model of liver metastasis After 10 weeks, mice were sacrificed.

Techniques: Injection, Control, Staining, Expressing