Journal: International Journal of Molecular Sciences

Article Title: Expression Profiles of Dopamine-Related Genes and miRNAs Regulating Their Expression in Breast Cancer

doi: 10.3390/ijms25126546

Figure Lengend Snippet: Venn diagram. DRD2 , dopamine receptor 2; DRD3 , dopamine receptor 3; DRD5 , dopamine receptor 5; TGF-β-2 , transforming growth factor beta 2; CAV2 , caveolin 2; SLC22A1 , solute carrier family 22 (organic cation transporter), member 1; CXCL1 2, chemokine (C-X-C motif) ligand 12; DRD1 , dopamine receptor 1; NR4A2 , nuclear receptor subfamily 4, group A, member 2; HRH2 , histamine receptor H2; NSG1 , neuron-specific gene family member 1.

Article Snippet: To accurately quantify protein concentrations, we used specific ELISA kits in accordance with the manufacturer’s instructions: Human Dopamine Receptor D5 Kit (MyBioSource, Inc., San Diego, CA, USA, Cat No. MBS724527), Human Dopamine Receptor D2 Kit (MyBioSource, Inc., San Diego, CA, USA, Cat No. MBS723432), Human Dopamine Receptor D3 Kit (MyBioSource, Inc., San Diego, CA, USA, Cat No. MBS722010), Human Caveolin 2 kit (MyBioSource, Inc., San Diego, CA, USA, Cat No. MBS1607726) and Human TGF beta 2 Kit (MyBioSource, Inc., San Diego, CA, USA, Cat No. MBS824902).

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Expression Profiles of Dopamine-Related Genes and miRNAs Regulating Their Expression in Breast Cancer

doi: 10.3390/ijms25126546

Figure Lengend Snippet: Changes in the expression profile of genes differentiating tumor samples compared to control tissues (−3.0 < FC > 3.0; p < 0.05).

Article Snippet: To accurately quantify protein concentrations, we used specific ELISA kits in accordance with the manufacturer’s instructions: Human Dopamine Receptor D5 Kit (MyBioSource, Inc., San Diego, CA, USA, Cat No. MBS724527), Human Dopamine Receptor D2 Kit (MyBioSource, Inc., San Diego, CA, USA, Cat No. MBS723432), Human Dopamine Receptor D3 Kit (MyBioSource, Inc., San Diego, CA, USA, Cat No. MBS722010), Human Caveolin 2 kit (MyBioSource, Inc., San Diego, CA, USA, Cat No. MBS1607726) and Human TGF beta 2 Kit (MyBioSource, Inc., San Diego, CA, USA, Cat No. MBS824902).

Techniques: Expressing

Journal: International Journal of Molecular Sciences

Article Title: Expression Profiles of Dopamine-Related Genes and miRNAs Regulating Their Expression in Breast Cancer

doi: 10.3390/ijms25126546

Figure Lengend Snippet: Changes in the expression profile of mRNAs in different breast cancer subtype samples compared to control samples obtained via RT-PCR. DRD2 , dopamine receptor 2; DRD3 , dopamine receptor 3; DRD5 , dopamine receptor 5; TGF-β-2 , transforming growth factor beta 2; CAV2 , caveolin 2. Data are presented as mean ± standard deviation.

Article Snippet: To accurately quantify protein concentrations, we used specific ELISA kits in accordance with the manufacturer’s instructions: Human Dopamine Receptor D5 Kit (MyBioSource, Inc., San Diego, CA, USA, Cat No. MBS724527), Human Dopamine Receptor D2 Kit (MyBioSource, Inc., San Diego, CA, USA, Cat No. MBS723432), Human Dopamine Receptor D3 Kit (MyBioSource, Inc., San Diego, CA, USA, Cat No. MBS722010), Human Caveolin 2 kit (MyBioSource, Inc., San Diego, CA, USA, Cat No. MBS1607726) and Human TGF beta 2 Kit (MyBioSource, Inc., San Diego, CA, USA, Cat No. MBS824902).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Standard Deviation

Journal: International Journal of Molecular Sciences

Article Title: Expression Profiles of Dopamine-Related Genes and miRNAs Regulating Their Expression in Breast Cancer

doi: 10.3390/ijms25126546

Figure Lengend Snippet: The expression profile of miRNAs potentially regulated by selected mRNAs in different breast cancer subtype tissues in comparison to control tissues.

Article Snippet: To accurately quantify protein concentrations, we used specific ELISA kits in accordance with the manufacturer’s instructions: Human Dopamine Receptor D5 Kit (MyBioSource, Inc., San Diego, CA, USA, Cat No. MBS724527), Human Dopamine Receptor D2 Kit (MyBioSource, Inc., San Diego, CA, USA, Cat No. MBS723432), Human Dopamine Receptor D3 Kit (MyBioSource, Inc., San Diego, CA, USA, Cat No. MBS722010), Human Caveolin 2 kit (MyBioSource, Inc., San Diego, CA, USA, Cat No. MBS1607726) and Human TGF beta 2 Kit (MyBioSource, Inc., San Diego, CA, USA, Cat No. MBS824902).

Techniques: Expressing, Comparison

Journal: International Journal of Molecular Sciences

Article Title: Expression Profiles of Dopamine-Related Genes and miRNAs Regulating Their Expression in Breast Cancer

doi: 10.3390/ijms25126546

Figure Lengend Snippet: The concentration of selected proteins related to the dopaminergic system in different subtypes of breast cancer and control tissue.

Article Snippet: To accurately quantify protein concentrations, we used specific ELISA kits in accordance with the manufacturer’s instructions: Human Dopamine Receptor D5 Kit (MyBioSource, Inc., San Diego, CA, USA, Cat No. MBS724527), Human Dopamine Receptor D2 Kit (MyBioSource, Inc., San Diego, CA, USA, Cat No. MBS723432), Human Dopamine Receptor D3 Kit (MyBioSource, Inc., San Diego, CA, USA, Cat No. MBS722010), Human Caveolin 2 kit (MyBioSource, Inc., San Diego, CA, USA, Cat No. MBS1607726) and Human TGF beta 2 Kit (MyBioSource, Inc., San Diego, CA, USA, Cat No. MBS824902).

Techniques: Concentration Assay

Journal: International Journal of Molecular Sciences

Article Title: Expression Profiles of Dopamine-Related Genes and miRNAs Regulating Their Expression in Breast Cancer

doi: 10.3390/ijms25126546

Figure Lengend Snippet: Overall survival analysis for DRD2 in every group.

Article Snippet: To accurately quantify protein concentrations, we used specific ELISA kits in accordance with the manufacturer’s instructions: Human Dopamine Receptor D5 Kit (MyBioSource, Inc., San Diego, CA, USA, Cat No. MBS724527), Human Dopamine Receptor D2 Kit (MyBioSource, Inc., San Diego, CA, USA, Cat No. MBS723432), Human Dopamine Receptor D3 Kit (MyBioSource, Inc., San Diego, CA, USA, Cat No. MBS722010), Human Caveolin 2 kit (MyBioSource, Inc., San Diego, CA, USA, Cat No. MBS1607726) and Human TGF beta 2 Kit (MyBioSource, Inc., San Diego, CA, USA, Cat No. MBS824902).

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Expression Profiles of Dopamine-Related Genes and miRNAs Regulating Their Expression in Breast Cancer

doi: 10.3390/ijms25126546

Figure Lengend Snippet: The sequence of primers used in qRT-PCR.

Article Snippet: To accurately quantify protein concentrations, we used specific ELISA kits in accordance with the manufacturer’s instructions: Human Dopamine Receptor D5 Kit (MyBioSource, Inc., San Diego, CA, USA, Cat No. MBS724527), Human Dopamine Receptor D2 Kit (MyBioSource, Inc., San Diego, CA, USA, Cat No. MBS723432), Human Dopamine Receptor D3 Kit (MyBioSource, Inc., San Diego, CA, USA, Cat No. MBS722010), Human Caveolin 2 kit (MyBioSource, Inc., San Diego, CA, USA, Cat No. MBS1607726) and Human TGF beta 2 Kit (MyBioSource, Inc., San Diego, CA, USA, Cat No. MBS824902).

Techniques: Sequencing

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

Article Title: Role of Dopamine Receptors in the Anticancer Activity of ONC201.

doi: 10.1016/j.neo.2017.10.002

Figure Lengend Snippet: Figure 1. D2-like receptors are expressed and functional in colorectal cancer cells. (A) Assessment of DRD2 mRNA expression in different cancer tissue samples using cBioportal. (B) Viability assessment after 72 hours of treatment with dopamine or sumanirole, as indicated in legend. Western blot analyses of PKA substrate phosphorylation in (C) HCT116 cells treated with 500 μM phosphodiesterase inhibitor IBMX and 10 μM adenylyl cyclase activator forskolin (F), or treated with 10 μM ONC201, in the presence or absence of 1 μM protein kinase A inhibitor peptide 6-22 (PKI); (D) Hep3B cells treated with different doses of ONC201 for 24 hours; (E) HCT116, RKO, and ONC201-resistant RKO cells (RKOr1) treated with 10 μM (HCT116) or 5 μM (RKOp and RKOr1) ONC201 for indicated times.

Article Snippet: Five different lentiviral plasmid shRNA constructs for DRD2 (Origene) were amplified, and DNA was isolated using plasmid mini kit (Qiagen) according to manufacturer's instructions.

Techniques: Functional Assay, Expressing, Western Blot, Phospho-proteomics

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

Article Title: Role of Dopamine Receptors in the Anticancer Activity of ONC201.

doi: 10.1016/j.neo.2017.10.002

Figure Lengend Snippet: Figure 3. Transient knockdown of DRD2 in colorectal cancer cells can activate the ISR and reduce cell number. (A) Viable cell count, (B) Western blot analyses of PARP cleavage and CHOP expression, (C) Western blot analyses of ATF4 and CHOP protein expression in cells after DRD2 siRNA-mediated knockdown for 24 hours and subsequent treatment with 10 μM ONC201 for indicated times. *P b .05 versus vehicle-treated scramble siRNA- transfected; P b .05 versus vehicle-treated DRD2 siRNA-transfected. (D) qRT-PCR analyses of DRD1 and DRD2 mRNA expression were performed to verify knockdown and monitor for potential compensatory overexpression of DRD1 receptor. Data are means ± SE from three biological replicates. P b .05 versus viability of control shRNA cells similarly treated. (E) Western blot analyses for PKA substrate phosphorylation in stably transfected control and DRD2 shRNA cells treated with 10 μM ONC201 for 24 hours. (F) Cell proliferation rate assessment of control and DRD2 shRNA-transfected cells was performed by enumerating cell number after indicated times of cell culture. Data are means ± SE from three biological replicates. P b .05 versus viability of control shRNA cells similarly treated.

Article Snippet: Five different lentiviral plasmid shRNA constructs for DRD2 (Origene) were amplified, and DNA was isolated using plasmid mini kit (Qiagen) according to manufacturer's instructions.

Techniques: Knockdown, Cell Counting, Western Blot, Expressing, Transfection, Quantitative RT-PCR, Over Expression, Control, shRNA, Phospho-proteomics, Stable Transfection, Cell Culture

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

Article Title: Role of Dopamine Receptors in the Anticancer Activity of ONC201.

doi: 10.1016/j.neo.2017.10.002

Figure Lengend Snippet: Figure 4. The anticancer effects of ONC201 are distinct from other selective DRD2 antagonists. (A) CTG assay images, (B) apoptosis measured by sub-G1 analyses, and Western blot analyses of (C) PARP cleavage and (D) ATF4 protein expression in different cancer cell types treated with different selective DRD2 antagonists (O: 10 μM ONC201 for HCT116, SKNSH, and U251, and 5 μM ONC201 for RKO; L: 25 μM L741626; R: 10 μM). (E) Nontumorigenic CCD-CON and IMR90 cells and HCT116 and SKNSH cancer cells treated with indicated doses of L-741,626 for 72 hours. *P b .05 versus vehicle treated; P b .05 versus L-741,626 treated.

Article Snippet: Five different lentiviral plasmid shRNA constructs for DRD2 (Origene) were amplified, and DNA was isolated using plasmid mini kit (Qiagen) according to manufacturer's instructions.

Techniques: CTG Assay, Western Blot, Expressing

Journal: bioRxiv

Article Title: Targeted Eicosanoid Profiling Reveals MicroRNA-155 Shifts PGE2/PGD2 Balance Towards Oncogenic State

doi: 10.1101/2020.04.08.032136

Figure Lengend Snippet: A. PGE2/PGD2 ratio in miR-155-positive (Lav670 and Lav714) and miR-155-depleted (Lav670+miRZIP155 and Lav714+miRZIP155) tumors (n=20). B-C. PGE2(B) and PGD2(C) level in miR-155 KO/KO and in miR-155 KO/+ tumors measured by ELISA (n=10). D-G. Relative RNA expression of Ptges1 (D), Ptges2 (E), Ptgds (F) and 15-Pgdh (G) in miR-155 KO/KO and miR-155 KO/+ tumors (n=10). H. Western blot analysis of key metabolic enzymes of PGE2/PGD2 production in miR-155 KO/KO and miR-155 KO/+ tumors (n=10). COX1, COX2, PTGES1, PTGES2, PTGDS, and 15-PGDH expressions are shown. β-actin antibody was used to ensure equal loading.

Article Snippet: The plasma concentrations of PGE2 and PGD2 were determined using ELISA kits from ENZO Life Sciences (Farmingdale, New York, NY, USA) and CUSABIO Biotechnology (Wu Han, China), respectively.

Techniques: Enzyme-linked Immunosorbent Assay, RNA Expression, Western Blot

Journal: bioRxiv

Article Title: Targeted Eicosanoid Profiling Reveals MicroRNA-155 Shifts PGE2/PGD2 Balance Towards Oncogenic State

doi: 10.1101/2020.04.08.032136

Figure Lengend Snippet: A. Relative PGE2 (left) and PGD2 (right) levels in MDA-MB-436 and Hs-578T cells in control (in blue) or in miR-155-knockdown cells (miRZIP155 in red). B. Relative PGE2 (left) or PGD2 (right) level in MCF7 cells with overexpression of miR-155 (miRH, in yellow). C-F. Relative RNA expression of Ptges1 (C), Ptges2 (D), Ptgds(E) and 15-Pgdh (F) in MDA-MB-436, Hs-578T, and MCF7 cells after knockdown (miRZIP155, in red) or overexpression (miRH155, in yellow) of miR-155. G. Western blot analysis of key metabolic enzymes for PGE2/PGD2 production in three breast cancer cell lines with after knockdown (miRZIP155) or overexpression (miRH155) of miR-155. β-actin antibody was used to ensure equal loading

Article Snippet: The plasma concentrations of PGE2 and PGD2 were determined using ELISA kits from ENZO Life Sciences (Farmingdale, New York, NY, USA) and CUSABIO Biotechnology (Wu Han, China), respectively.

Techniques: Control, Knockdown, Over Expression, RNA Expression, Western Blot

Journal: bioRxiv

Article Title: Targeted Eicosanoid Profiling Reveals MicroRNA-155 Shifts PGE2/PGD2 Balance Towards Oncogenic State

doi: 10.1101/2020.04.08.032136

Figure Lengend Snippet: A. Relative level of miR-155 in six PDCs. Three high-expressers were labeled as PDCH and low-expressers were labeled as PDCL. Yellow bars on PDCL (miRH 155) indicates miR-155 level expressed by lentiviral infection. B and C. The level of PGE2(B) or PGD2(C) in three PDCLs with overexpressed miR-155 (miRH155, in yellow). D-G. Relative RNA expression of Ptges1 (D), Ptges2 (E), Ptgds (F) and 15-Pgdh (G) in PDCH, PDCL, and PDCL with lentiviral miR-155 expression (miRH155). H. Western blot analysis of key metabolic enzymes for PGE2/PGD2 production in three PDCH, PDCL, and PDCL with miR-155 expression (marked as +). β-actin antibody was used to ensure equal loading.

Article Snippet: The plasma concentrations of PGE2 and PGD2 were determined using ELISA kits from ENZO Life Sciences (Farmingdale, New York, NY, USA) and CUSABIO Biotechnology (Wu Han, China), respectively.

Techniques: Labeling, Infection, RNA Expression, Expressing, Western Blot

Journal: bioRxiv

Article Title: Targeted Eicosanoid Profiling Reveals MicroRNA-155 Shifts PGE2/PGD2 Balance Towards Oncogenic State

doi: 10.1101/2020.04.08.032136

Figure Lengend Snippet: A. Correlation analysis between miR-155 (in blue) and PGE2/PGD2 ratio (in red) in human TNBC specimens. p=0.045, B-E. Relative RNA expression of PTGES1 (b), PTGES2 (c), PTGDS (d) and 15-PGDH (e) in miR-155-high (H1-H10, in red) and miR-155-low (L1-L10, in blue) TNBC samples. F. Western blot results of key metabolic enzymes and suggested novel regulators for PGE2/PGD2 metabolism in the miR-155-high and -low TNBC specimens. Results of COX1, COX2, PTGES1, PTGES2, PTGDS, 15-PGDH, cMYC, and KLF4 are shown. β-actin was used to ensure equal loading. G and H. Correlation analysis between miR-155 and PGE2(g) or PGD2(h) in the plasma samples of selected TNBC patients. I. Overall survival curves of breast cancer patients according to the levels of cMYC(upper left), KLF4(upper right), PTGES2(lower left) and PTGDS(lower right), using the ProgeneV2 webtool. J. Graphical abstract showing the molecular mechanism of PGD2/PGE2 production regulated by miR-155.

Article Snippet: The plasma concentrations of PGE2 and PGD2 were determined using ELISA kits from ENZO Life Sciences (Farmingdale, New York, NY, USA) and CUSABIO Biotechnology (Wu Han, China), respectively.

Techniques: RNA Expression, Western Blot, Clinical Proteomics

Journal: bioRxiv

Article Title: Abnormal eNK cells contribute to endometrial fibrosis in intrauterine adhesions patients

doi: 10.1101/2024.12.01.626271

Figure Lengend Snippet: (A) Bubble diagram showing the top10 DEGs of eNK cells between normal controls and IUA patients based on scRNA-seq analysis. (B) Immunofluorescence staining of CD56 (green) and PTGDS (red) in endometria of normal controls (n = 6) and IUA patients (n = 6) and the relative optical densities (OD) of PTGDS staining in CD56+ cells analyzed by Image J. Scale bars: 50 μm. (C) The level of PGD2 in the supernatants of eNK cells (n = 15) and pbNK cells (n = 6). (D-H) The ESCs were pre-treated with 10 ng/ml TGFβ1 for 48 h and then treated with 10 μM PGD2 for 48 h. (D-F) The mRNA levels of ACTA2, Col-1 and FN1 in ESCs (n = 4) examined by qRT-PCR. (G) Left: The protein levels of α-SMA and Col-1 in ESCs (n = 3) examined by Western Blotting. Right: Relative band intensities analyzed by Image J. (H) Immunofluorescence staining of α-SMA and Col-1 in ESCs (n = 5) and their relative optical densities (OD) were analyzed by Image J. Scale bars: 100 μm. (I-M) The ESCs were treated with 1 μM PGD2-IN-1 24 h before eNK cells incubation. (I-K) The mRNA levels of ACTA2, Col-1 and FN1 in ESCs (n = 4) examined by qRT-PCR. (L) The protein levels of α-SMA, Col-1 and FN1 in ESCs (n = 3) examined by Western Blotting. (M) Relative band intensities of α-SMA, Col-1 and FN1 analyzed by Image J. Error bars, mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: The levels of PGD2 in the supernatant of eNK and pbNK cells were measured with a commercial human PGD2 ELISA kit (Elabscience) according to the manufacturer’s instruction.

Techniques: Immunofluorescence, Staining, Quantitative RT-PCR, Western Blot, Incubation

Journal: bioRxiv

Article Title: Abnormal eNK cells contribute to endometrial fibrosis in intrauterine adhesions patients

doi: 10.1101/2024.12.01.626271

Figure Lengend Snippet: (A) Venn analysis of the genes upregulated in myofibroblasts, downregulated in eNK-treated myofibroblasts and transcription factors associated with myofibroblasts. (B) Heatmap of genes (BNC2, OSR1, ZNF521, FOS and BCL11A) in ESCs, myofibroblasts and eNK-treated myofibroblasts. (C) The mRNA level of ZNF521 in ESCs (n = 5) detected by qRT-PCR. (D-E) Top: The protein level of ZNF521 in ESCs (n = 3). Bottom: Relative band intensities analyzed by Image J. (F) The mRNA level of ZNF521 in endometrial of normal controls (n = 25) and IUA patients (n = 25) detected by qRT-PCR. (G) Immunohistochemical staining of ZNF521 in endometria of normal controls (n = 6) and IUA patients (n = 6). Scale bars: 50 μm. (H) The mRNA levels of ACTA2, Col-1 and FN1 in ESCs (n = 3, pre-treated with TGFβ1) transfected with si-NC or si-ZNF521 for 48 h examined by qRT-PCR. (I) Top: The protein levels of α-SMA and Col-1 in ESCs (n = 3) detected by Western Blotting. Bottom: Relative band intensities analyzed by Image J. (J) Top: The protein levels of α-SMA and Col-1 in ESCs (n = 3, pre-treated with eNK cells) transfected with ZNF521 plasmid for 48 h detected by Western Blotting. Bottom: Relative band intensities analyzed by Image J. (K) Top: The protein levels of α-SMA and Col-1 in ESCs (n = 3, pre-treated with PGD2) transfected with ZNF521 plasmid detected by Western Blotting. Bottom: Relative band intensities analyzed by Image J. Error bars, mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: The levels of PGD2 in the supernatant of eNK and pbNK cells were measured with a commercial human PGD2 ELISA kit (Elabscience) according to the manufacturer’s instruction.

Techniques: Quantitative RT-PCR, Immunohistochemical staining, Staining, Transfection, Western Blot, Plasmid Preparation

Journal: bioRxiv

Article Title: Abnormal eNK cells contribute to endometrial fibrosis in intrauterine adhesions patients

doi: 10.1101/2024.12.01.626271

Figure Lengend Snippet: (A) The mice were divided into the following groups: Sham operation (n = 6), curettage (n = 6), curettage + supernatants of eNK cells (n = 6), curettage + PGD2 (200 ug/kg, n = 6), curettage + supernatants of eNK cells + PGD2-IN-1 (250 ug/kg, n = 6), curettage + supernatants of pbNK cells (n = 6). (B) HE (scale bars of HE: 100 μm), Masson staining and immunohistochemical staining of α-SMA, Col-1 and ZNF521 in endometria of mice. Scale bars: 50 μm. (C) The relative AOD of Masson, α-SMA, Col-1 and ZNF521 analyzed by Image J. Error bars, mean ± SD. *** P < 0.001, **** P < 0.0001, ns: not significant.

Article Snippet: The levels of PGD2 in the supernatant of eNK and pbNK cells were measured with a commercial human PGD2 ELISA kit (Elabscience) according to the manufacturer’s instruction.

Techniques: Staining, Immunohistochemical staining