Journal: Oncogene

Article Title: Olfactomedin 4 deletion induces colon adenocarcinoma in Apc Min/+ mice

doi: 10.1038/onc.2016.58

Figure Lengend Snippet: Olfm4 deletion induces large polyps in the small intestine and adenocarcinoma formation in the distal colon of Apc +/− mice. ( a ) Polyp formation (arrow) in the small intestine of Apc Min/+ , Apc +/− Olfm4 +/− and Apc +/− Olfm4 −/− ( Apc Olfm4 double-mutant) mice. ( b ) Adenocarcinoma formation in the distal colon of Apc +/− Olfm4 +/− and Apc Olfm4 double-mutant mice. Three representative tumors in mice of each genotype are presented. Hematoxylin and eosin (HE) histology staining is shown below each corresponding tumor. Scale bar, 50 μm. ( c ) Mean (±s.d.) small intestinal polyp number and size, and colon-tumor incidence in Apc Min/+ ( n =20), Apc +/− Olfm4 +/− ( n =23) and Apc Olfm4 double-mutant mice ( n =22) mice.

Article Snippet: Human OLFM4 protein was purchased from Sino Biological Inc. (Beijing, China).

Techniques: Mutagenesis, Staining

Journal: Oncogene

Article Title: Olfactomedin 4 deletion induces colon adenocarcinoma in Apc Min/+ mice

doi: 10.1038/onc.2016.58

Figure Lengend Snippet: Olfm4 deletion enhances Wnt/β-catenin signaling in Apc Olfm4 double-mutant mice. ( a–h ) β-Catenin immunohistochemistry staining was performed in the distal colon of Apc +/+ Olfm4 +/+ (WT) ( a ), Apc +/− Olfm4 +/+ ( b ), Apc +/+ Olfm4 −/− ( c ), Apc +/− Olfm4 +/− ( d ) and Apc +/− Olfm4 −/− ( e and g ) mice. Hematoxylin and eosin (HE) staining in ( f and h ) was performed in mice corresponding to ( e and g ), respectively. Scale bar, 50 μm. ( i ) A heatmap from cDNA microarray analysis shows Wnt pathway-related or target gene-expression level in the distal colon of mice with different genotypes. N, tumor-adjacent normal tissue; T, tumor. ( j ) Mean (±s.d.) ( n =3) gene-expression levels for six representative Wnt target genes were confirmed by qRT–PCR. See also and .

Article Snippet: Human OLFM4 protein was purchased from Sino Biological Inc. (Beijing, China).

Techniques: Mutagenesis, Immunohistochemistry, Staining, Microarray, Expressing, Quantitative RT-PCR

Journal: Oncogene

Article Title: Olfactomedin 4 deletion induces colon adenocarcinoma in Apc Min/+ mice

doi: 10.1038/onc.2016.58

Figure Lengend Snippet: OLFM4 decreases β-catenin levels. ( a–k ) Western blots were performed in 293T cells ( a–i ), SW480 cells ( j ) and freshly isolated mouse colon crypts ( k ) after human OLFM4 treatment (1 μg/ml) ( a–j ) and mouse Olfm4 treatment (1 μg/ml) ( k ). β-Actin, total cadherin, Hsp90 and SP1 were used as loading controls in total cell lysate, membrane, cytoplasm and nucleus cell preparations, respectively. ( a ) β-Catenin levels in total cell lysate, membrane, cytoplasm and nucleus at different time points of OLFM4 treatment. ( b ) Endogenous and Wnt3a (1 μg/ml)-induced β-catenin levels in the cytoplasm and nucleus of cells pretreated with and without OLFM4 for 30 min. ( c ) Endogenous and Wnt3a (1 μg/ml)-induced β-catenin levels in the cytoplasm of cells transfected with control (vector) or OLFM4 expression plasmid. ( d ) β-Catenin phosphorylation at different time points of OLFM4 treatment. ( e and f ) Akt (T308 or Ser473) phosphorylation at different time points of OLFM4 treatment. ( g and h ) β-Catenin levels in the cytoplasm of cells pretreated with and without either LiCl (20 m m ) or MG132 (10 μ m ) before OLFM4 treatment for 30 min. ( i ) β-Catenin levels in total cell lysates of cells transfected with Flag-tagged WT or mutant (S33Y or S31/37T41S45) β-catenin expression plasmids and then treated with or without OLFM4 for 30 min. ( j ) Total and active β-catenin levels in the cytoplasm or nucleus of SW480 colon-cancer cells at different time points of Olfm4 treatment. ( k ) β-Catenin levels in the cytoplasm and nucleus of freshly isolated mouse colon crypts at different time points of Olfm4 (or PBS control buffer) treatment. ( l ) Olfm4 and β-catenin levels in the cytoplasm of Ls174T cells infected with lentiviral control shRNA and Olfm4 shRNA.

Article Snippet: Human OLFM4 protein was purchased from Sino Biological Inc. (Beijing, China).

Techniques: Western Blot, Isolation, Transfection, Plasmid Preparation, Expressing, Mutagenesis, Infection, shRNA

Journal: Oncogene

Article Title: Olfactomedin 4 deletion induces colon adenocarcinoma in Apc Min/+ mice

doi: 10.1038/onc.2016.58

Figure Lengend Snippet: OLFM4 interacts with Frizzled-7 and -10. ( a ) Ls174T cell lysate in the presence or absence of OLFM4 treatment was immunoprecipitated with OLFM4 antibody or IgG control antibody and then subjected to western blot with a total Frizzled (Fzd) receptors antibody. ( b ) Ls174T cell lysate was immunoprecipitated with OLFM4 antibody or IgG control antibody and then subjected to western blot with Fzd-7 or Fzd-10 antibody. ( c ) Ls174T cell lysate was immunoprecipitated with a total Fzd receptors antibody, Fzd-10 antibody or IgG antibody and then subjected to western blot with OLFM4 antibody. ( d ) Cell lysates from 293T cells stably overexpressing OLFM4 were immunoprecipitated with OLFM4 antibody or IgG control antibody and then subjected to western blot with a total Fzd receptors antibody, Fzd-7 antibody or Fzd-10 antibody. ( e ) Cell lysate from 293T cells stably overexpressing OLFM4 was immunoprecipitated with a total Fzd receptors antibody, Fzd-7 antibody, Fzd-10 antibody or IgG control antibody and then subjected to western blot with OLFM4 antibody. ( f ) Microscale thermophoresis (MST) measurements of the OLFM4 interaction with Fzd-7 (solid circles) and Fzd-10 (hollow circles). The solid lines are the best-fit curves to the data using the Hill equation ( K D =2.5 μ m for the OLFM4–Fzd-7 interaction and K D =10 μ m for the OLFM4–Fzd-10 interaction). The error bars represent the s.d. from three independent measurements.

Article Snippet: Human OLFM4 protein was purchased from Sino Biological Inc. (Beijing, China).

Techniques: Immunoprecipitation, Western Blot, Stable Transfection, Microscale Thermophoresis

Journal: Oncogene

Article Title: Olfactomedin 4 deletion induces colon adenocarcinoma in Apc Min/+ mice

doi: 10.1038/onc.2016.58

Figure Lengend Snippet: OLFM4 is a target gene of the Wnt/β-catenin/TCF pathway. ( a ) Three LEF1/TCF binding sites (in black box) located in the promoter of OLFM4 . ( b ) The core (CAAAG or CTTTG) of the LEF1/TCF binding site was deleted individually or in combination as indicated by gray boxes. Data represent mean (±s.d.) ( n =3) relative luciferase activity of corresponding reporter in the presence of Wnt3a or vehicle in 293T cells. * P< 0.05 and ** P< 0.01 when compared with WT vehicle or WT Wnt3a, respectively, two-tailed t -test. ( c ) Mean (±s.d.) ( n =3) OLFM4 mRNA expression determined by qRT–PCR in Ls174T cells (right) after knockdown of TCF4 expression by TCF4 lentivirus shRNA (left). * P< 0.01 when compared with control (Con) shRNA, two-tailed t -test. ( d ) Mean (±s.d.) ( n =3) OLFM4 mRNA expression determined by qRT–PCR in Ls174T and SW948 cells treated with IWP2, XAV-939 or DMSO control. * P< 0.01 when compared with DMSO group, two-tailed t -test. ( e ) ChIP assay of LEF1 and TCF4 binding to all three LEF/TCF binding sites in the OLFM4 promoter.

Article Snippet: Human OLFM4 protein was purchased from Sino Biological Inc. (Beijing, China).

Techniques: Binding Assay, Luciferase, Activity Assay, Two Tailed Test, Expressing, Quantitative RT-PCR, shRNA

Journal: Oncogene

Article Title: Olfactomedin 4 deletion induces colon adenocarcinoma in Apc Min/+ mice

doi: 10.1038/onc.2016.58

Figure Lengend Snippet: Olfm4 deletion results in intestinal-crypt hyperplasia and severe inflammation after AOM/DSS treatment. ( a ) Schematic of AOM/DSS-induced colitis-associated cancer model. Mice were injected intraperitoneally with AOM (10 mg/kg). After 5 days, mice were treated with 2.5% DSS in their drinking water for 5 days followed by regular water for 16 days. The cycle was repeated four times. ( b ) WT ( n =8) and Olfm4 −/− ( n =8) mice were weighed after four cycles of DSS and water treatment. * P< 0.01, two-tailed t -test. ( c ) Percentage of survival of WT ( n =10) and Olfm4 −/− ( n =10) mice after four cycles of DSS and water treatment. ( d ) Length of colon in WT ( n =5) and Olfm4 −/− ( n =5) mice after four cycles of DSS and water treatment. * P< 0.01, two-tailed t -test. ( e ) Mean (±s.d.) histology score of inflammation and tissue damage in the jejunum, ileum and colon of WT ( n =8) and Olfm4 −/− ( n =8) mice after four cycles of DSS and water treatment. * P< 0.01 when compared with WT corresponding tissues, two-tailed t -test. ( f ) HE staining of jejunum, ileum and colon of WT and Olfm4 −/− mice after four cycles of DSS and water treatment. Scale bar, 50 μm. The image represents at least five mice in each group.

Article Snippet: Human OLFM4 protein was purchased from Sino Biological Inc. (Beijing, China).

Techniques: Injection, Two Tailed Test, Staining

Journal: Breast Cancer Research : BCR

Article Title: Oncogene activated human breast luminal progenitors contribute basally located myoepithelial cells

doi: 10.1186/s13058-024-01939-x

Figure Lengend Snippet: Antibody list

Article Snippet: OLFM4 , – , Atlas antibodies , HPA077718 , 1:1000; C.

Techniques:

Journal: International Journal of Oncology

Article Title: High levels of human epididymis protein 4 mRNA and protein expression are associated with chemoresistance and a poor prognosis in pancreatic cancer

doi: 10.3892/ijo.2020.5147

Figure Lengend Snippet: Identification of chemotherapy resistance molecules. (A and B) Results of NGS analysis of (A) GEM and (B) GEM + nab-PTX administration. The genes are shown on the horizontal axis, and the treatment resistance score is shown on the vertical axis. Treatment resistance score was calculated by the ratio of NE value (treated group/control group) multiplied by the difference in NE value (treated group-control group). In both cases (A and B), HE4 was upregulated in the treatment group. (C and D) The NE value of HE4 mRNA. The NE value ratio of HE4 expression between the treatment and control groups was often >1.0 in PDXs treated with GEM or GEM plus nab-PTX. HE4, human epididymis protein 4; PDX, patient-derived xenograft; GEM, gemcitabine; nab-PTX, nab-paclitaxel.

Article Snippet: For transient overexpression experiments, an expression plasmid vector for HE4 (pCMV6-AC-HE4-GFP tag plasmid, NM_006418) was obtained from OriGene Technologies, Inc. and an empty vector (pCMV6-AC-GFP) was adopted for control transfection.

Techniques: Expressing, Derivative Assay

Journal: International Journal of Oncology

Article Title: High levels of human epididymis protein 4 mRNA and protein expression are associated with chemoresistance and a poor prognosis in pancreatic cancer

doi: 10.3892/ijo.2020.5147

Figure Lengend Snippet: Kaplan-Meier plots demonstrating the correlation between HE4 mRNA expression and patient survival in TCGA database for pancreatic cancer (n=166). Red line, high expression (n=34); blue line, low expression (n=132). HE4, human epididymis protein 4. The asterisk (*) indicates a statistically significant difference (P<0.05).

Article Snippet: For transient overexpression experiments, an expression plasmid vector for HE4 (pCMV6-AC-HE4-GFP tag plasmid, NM_006418) was obtained from OriGene Technologies, Inc. and an empty vector (pCMV6-AC-GFP) was adopted for control transfection.

Techniques: Expressing

Journal: International Journal of Oncology

Article Title: High levels of human epididymis protein 4 mRNA and protein expression are associated with chemoresistance and a poor prognosis in pancreatic cancer

doi: 10.3892/ijo.2020.5147

Figure Lengend Snippet: Expression of HE4 and cell viability assays of cell lines. (A) The levels of HE4 gene expression in 9 tumor cell lines (SK-N-AS, KATOIII, ASPC-1, PANC-1, SUIT-2, MIA-Paca2, IMR-32, HCT116 and HeLa) and 2 non-cancerous cell lines (HUVECs and HRGECs) measured by RT-qPCR. Endogenous HE4 expression levels differed among cell lines. * P<0.05. (B) Cell viability assays of tumor cell lines in response to GEM treatment. When the concentration of GEM was increased, the relative viable cell numbers tended to be higher in the cell lines in which expression of endogenous HE4 was relatively higher. Cell lines with different lowercase letters (a-g) indicate statistically significant differences at each GEM concentration. a P<0.05 vs. b-g; b P<0.05 vs. c-g; c P<0.05 vs. d-g; d P<0.05 vs. e-g; e P<0.05 vs. f and g; and f P<0.05 vs. g. HE4, human epididymis protein 4; GEM, gemcitabine; HUVECs, human umbilical vein endothelial cells; HRGECs, human renal glomerular endothelial cells.

Article Snippet: For transient overexpression experiments, an expression plasmid vector for HE4 (pCMV6-AC-HE4-GFP tag plasmid, NM_006418) was obtained from OriGene Technologies, Inc. and an empty vector (pCMV6-AC-GFP) was adopted for control transfection.

Techniques: Expressing, Quantitative RT-PCR, Concentration Assay

Journal: International Journal of Oncology

Article Title: High levels of human epididymis protein 4 mRNA and protein expression are associated with chemoresistance and a poor prognosis in pancreatic cancer

doi: 10.3892/ijo.2020.5147

Figure Lengend Snippet: Viability assay of 293 cells. (A) Expression of HE4 and control vectors in 293 cells followed by treatment with GEM after 24 h (day 1). MTT assay was performed 48 h following treatment with GEM at various concentrations (day 3). (B and C) Histograms representing the rates of change of each measured OD value of control vector and HE4-expressing 293 cells. * P<0.0007, control vector vs. HE4-expressing cells at each GEM concentration, as determined by two-way ANOVA for multiple comparisons with the Bonferroni post hoc test. HE4, human epididymis protein 4; GEM, gemcitabine.

Article Snippet: For transient overexpression experiments, an expression plasmid vector for HE4 (pCMV6-AC-HE4-GFP tag plasmid, NM_006418) was obtained from OriGene Technologies, Inc. and an empty vector (pCMV6-AC-GFP) was adopted for control transfection.

Techniques: Viability Assay, Expressing, MTT Assay, Plasmid Preparation, Concentration Assay

Journal: International Journal of Oncology

Article Title: High levels of human epididymis protein 4 mRNA and protein expression are associated with chemoresistance and a poor prognosis in pancreatic cancer

doi: 10.3892/ijo.2020.5147

Figure Lengend Snippet: Association between HE4 expression and patient prognosis. (A) IHC analysis of HE4 in patient tissues (magnification, ×100). Images are of the same sample tissue sections and representing the intensity of stain. Images in the left panel show H&E staining, and those in the right indicate IHC detection of HE4. (B) The criteria for evaluation of HE4 expression levels. The intensity of stain and percentage criteria are designated. (C) Kaplan-Meier survival analysis in pancreatic cancer patients (n=80). Overall survival is shown in accordance with the protein expression level of HE4. The red line represents the group of patients with high HE4 expression (n=44), and the blue line indicates those with low HE4 expression (n=36). HE4, human epididymis protein 4.

Article Snippet: For transient overexpression experiments, an expression plasmid vector for HE4 (pCMV6-AC-HE4-GFP tag plasmid, NM_006418) was obtained from OriGene Technologies, Inc. and an empty vector (pCMV6-AC-GFP) was adopted for control transfection.

Techniques: Expressing, Staining

Journal: International Journal of Oncology

Article Title: High levels of human epididymis protein 4 mRNA and protein expression are associated with chemoresistance and a poor prognosis in pancreatic cancer

doi: 10.3892/ijo.2020.5147

Figure Lengend Snippet: Association between HE4 expression and clinicopathological features in 80 cases of pancreatic cancer.

Article Snippet: For transient overexpression experiments, an expression plasmid vector for HE4 (pCMV6-AC-HE4-GFP tag plasmid, NM_006418) was obtained from OriGene Technologies, Inc. and an empty vector (pCMV6-AC-GFP) was adopted for control transfection.

Techniques: Expressing

Journal: International Journal of Oncology

Article Title: High levels of human epididymis protein 4 mRNA and protein expression are associated with chemoresistance and a poor prognosis in pancreatic cancer

doi: 10.3892/ijo.2020.5147

Figure Lengend Snippet: Univariate and multivariate analyses of prognostic factor for overall survival in 80 pancreatic cancer patients.

Article Snippet: For transient overexpression experiments, an expression plasmid vector for HE4 (pCMV6-AC-HE4-GFP tag plasmid, NM_006418) was obtained from OriGene Technologies, Inc. and an empty vector (pCMV6-AC-GFP) was adopted for control transfection.

Techniques:

Journal: Molecular Systems Biology

Article Title: A Notch positive feedback in the intestinal stem cell niche is essential for stem cell self‐renewal

doi: 10.15252/msb.20167324

Figure Lengend Snippet: Table of antibodies

Article Snippet: Anti‐OLFM4 , Sino Biological Inc. , 11639‐MM12‐50 , 1:1,000 (FC).

Techniques: