timp3 expression  (OriGene)

Journal: Cell communication and signaling : CCS

Article Title: Glycosaminoglycan modifications of betaglycan regulate ectodomain shedding to fine-tune TGF-β signaling responses in ovarian cancer.

doi: 10.1186/s12964-024-01496-y

Figure Lengend Snippet: Fig. 5 TIMP3 negatively regulates glycosaminoglycan-dependent BG ectodomain shedding. A Scatter plot of the differentially expressed genes from transcriptomics of HS-BG and ∆GAG-BG plotted against CS-BG and ∆GAG-BG comparison groups. All protease-related genes are highlighted. B Volcano plots of the differentially expressed genes between ∆GAG-BG vs. Control-vector, S534A (CS-BG) vs. ∆GAG-BG, and S545A (HS-BG) vs. ∆GAG-BG. (TIMP3; ∆GAG vs Cntl, log2FC = 0.524, -log(10)p = 1.643, ∆GAG vs CS-BG, log2FC = 1.054, -log(10)p = 2.493, ∆GAG vs HS-BG, log2FC = 0.364, -log(10)p = 1.1411.). C, E Semi-qRT-PCR of TIMP3 and (D, F) ELISA for sol-BG in indicated cells normalized to the scramble vector cells. Mean ± SEM (n = 3 qRT-PCR), *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001, One-way ANOVA followed by unpaired t-test between scramble vector treated cells to siTIMP3 treated cells. (HEYA8 p = 0.0055 ∆GAG-BG SCRM vs siTIMP3, SKOV-3 p = 0.0007, ∆GAG-BG SCRM compared to siTIMP3), for ELISA in (D, F) n = 5 HEYA8, n = 6 SKOV-3, p = 0.0068, ∆GAG-BG SCRM vs siTIMP3 in HEYA8 and p = < 0.0001, ∆GAG SCRM vs. siTIMP3 in SKOV-3

Article Snippet: Origene siRNA-27 kit was used for transient knockdown of TIMP3 expression (SR304839 Locus IF 7078).

Techniques: Comparison, Control, Plasmid Preparation, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: Advanced healthcare materials

Article Title: In Situ Deployment of Engineered Extracellular Vesicles into the Tumor Niche via Myeloid-Derived Suppressor Cells.

doi: 10.1002/adhm.202101619

Figure Lengend Snippet: Fig. 1. Non-viral transfection of MDSCs mediates the release of engineered EVs with desirable cargo. (a) Schematic diagram illustrating the experimental design. (1) MDSCs were electroporated with expression plasmids for Timp3 or Rarres2. Electroporation with sham/empty plasmids served as control. (2) The plasmids are expressed within the MDSCs and (3) transcripts are packed and released within EVs. qRT-PCR analysis of the MDSC cultures at (b) 12, (c) 24, (d) 48, and (e) 72 hours post-electroporation reveals strong Timp3 or Rarres2 overexpression. Analysis of the EVs isolated from the supernatant at (f) 24 and (g) 48 hours

Article Snippet: Plasmid vector Company Cat. No Backbone Sham Origene PS100001 pCMV6 Timp3 (Mouse tissue inhibitor metalloproteinase 3) Origene MG202295 pCMV6 Rarres2 (Mouse retinoic acid receptor responder) Origene MG222586 pCMV6 Adv Healthc Mater.

Techniques: Transfection, Expressing, Electroporation, Control, Quantitative RT-PCR, Over Expression, Isolation

Journal: Advanced healthcare materials

Article Title: In Situ Deployment of Engineered Extracellular Vesicles into the Tumor Niche via Myeloid-Derived Suppressor Cells.

doi: 10.1002/adhm.202101619

Figure Lengend Snippet: Fig. 2. MDSC-derived EVs can be internalized by cancer cells and modulate gene expression. (a) Schematic diagram illustrating the experimental design. MDSCs were electroporated with expression plasmids for Timp3 or Rarres2. Electroporation with sham/empty plasmids served as control. MDSC-derived EVs were isolated and incubated with Py8119 mouse breast cancer cell cultures for 6 – 24 hours. (b) Fluorescence microscopy imaging of the Py8119 cells (labeled red and blue) revealed successful uptake of MDSC-derived EVs (labeled green). The images shown represent EV uptake at 24 h post-exposure. qRT-PCR analysis of the Py8119 cultures at (c) 6, (d) 12, and (d) 24 hours post-EV exposure reveals strong Timp3 or Rarres2 overexpression after 12 hours of exposure. ** p< 0.01 (n= 4), **** p< 0.0001 (n= 4).

Article Snippet: Plasmid vector Company Cat. No Backbone Sham Origene PS100001 pCMV6 Timp3 (Mouse tissue inhibitor metalloproteinase 3) Origene MG202295 pCMV6 Rarres2 (Mouse retinoic acid receptor responder) Origene MG222586 pCMV6 Adv Healthc Mater.

Techniques: Derivative Assay, Gene Expression, Expressing, Electroporation, Control, Isolation, Incubation, Fluorescence, Microscopy, Imaging, Labeling, Quantitative RT-PCR, Over Expression

Journal: Advanced healthcare materials

Article Title: In Situ Deployment of Engineered Extracellular Vesicles into the Tumor Niche via Myeloid-Derived Suppressor Cells.

doi: 10.1002/adhm.202101619

Figure Lengend Snippet: Fig. 3. Transfected MDSCs can transfer engineered EVs to breast cancer cells, in situ, and mediate gene expression and cellular responses. (a) Schematic diagram illustrating the experimental design. MDSCs were electroporated with expression plasmids for Timp3 or Rarres2. Electroporation with sham/empty plasmids served as control. Transfected MDSCs and Py8119 mouse breast cancer cells were co- cultured using a transwell system, with the MDSCs in the apical chamber and Py8119 cells in the basal chamber. The MDSCs were-prelabeled with a membrane dye to trace EV release and uptake. MDSC-derived EVs were thus expected to translocate across the membrane

Article Snippet: Plasmid vector Company Cat. No Backbone Sham Origene PS100001 pCMV6 Timp3 (Mouse tissue inhibitor metalloproteinase 3) Origene MG202295 pCMV6 Rarres2 (Mouse retinoic acid receptor responder) Origene MG222586 pCMV6 Adv Healthc Mater.

Techniques: Transfection, In Situ, Gene Expression, Expressing, Electroporation, Control, Cell Culture, Membrane, Derivative Assay

Journal: Advanced healthcare materials

Article Title: In Situ Deployment of Engineered Extracellular Vesicles into the Tumor Niche via Myeloid-Derived Suppressor Cells.

doi: 10.1002/adhm.202101619

Figure Lengend Snippet: Fig. 4. Transfected MDSCs can also modulate cancer cell proliferation and invasion, in situ. (a) Schematic diagram illustrating the experimental design. MDSCs were electroporated with expression plasmids for Timp3 or Rarres2. Electroporation with sham/empty plasmids served as control. Transfected MDSCs and Py8119 mouse breast cancer cells were co- cultured in direct contact. Matrigel-coated transwell insets were used for cell invasion studies. The MDSCs were-prelabeled green, and the Py8119 cells were prelabeled red. (b) Fluorescence microscopy imaging of the MDSC/Py8119 co-cultures after 24 hours. (c) Flow cytometry analysis revealed reduced Py8119 cell numbers in co-cultured with Timp3- or

Article Snippet: Plasmid vector Company Cat. No Backbone Sham Origene PS100001 pCMV6 Timp3 (Mouse tissue inhibitor metalloproteinase 3) Origene MG202295 pCMV6 Rarres2 (Mouse retinoic acid receptor responder) Origene MG222586 pCMV6 Adv Healthc Mater.

Techniques: Transfection, In Situ, Expressing, Electroporation, Control, Cell Culture, Fluorescence, Microscopy, Imaging, Flow Cytometry

Journal: Advanced healthcare materials

Article Title: In Situ Deployment of Engineered Extracellular Vesicles into the Tumor Niche via Myeloid-Derived Suppressor Cells.

doi: 10.1002/adhm.202101619

Figure Lengend Snippet: Fig. 5. Transfected MDSCs retain tumor-homing abilities and drive anti-tumoral gene and protein expression. (a) Schematic diagram illustrating the experimental design. MDSCs were electroporated with expression plasmids for Timp3 or Rarres2. Electroporation with sham/empty plasmids served as control. Transfected MDSCs were then injected into tumor-bearing mice via the tail vein, and accumulation and gene/protein expression in the tumor was evaluated after 24 hours. (b, c) IVIS imaging after 24 hours revealed strong accumulation of transfected MDSCs in the tumor niche compared to other organs. Transfected MDSCs were

Article Snippet: Plasmid vector Company Cat. No Backbone Sham Origene PS100001 pCMV6 Timp3 (Mouse tissue inhibitor metalloproteinase 3) Origene MG202295 pCMV6 Rarres2 (Mouse retinoic acid receptor responder) Origene MG222586 pCMV6 Adv Healthc Mater.

Techniques: Transfection, Expressing, Electroporation, Control, Injection, Imaging

Journal: Frontiers in Genetics

Article Title: TIMP3 Overexpression Improves the Sensitivity of Osteosarcoma to Cisplatin by Reducing IL-6 Production

doi: 10.3389/fgene.2018.00135

Figure Lengend Snippet: Sequences of primers used in real-time PCR.

Article Snippet: For TIMP3 overexpression, Saos2-lung, MG63, and U2OS cells were transfected with green fluorescent protein (GFP)-labeled-lentiviral vectors (GeneChem, Shanghai, China) expressing TIMP3 or a vector with a scrambled control sequence.

Techniques:

Journal: Frontiers in Genetics

Article Title: TIMP3 Overexpression Improves the Sensitivity of Osteosarcoma to Cisplatin by Reducing IL-6 Production

doi: 10.3389/fgene.2018.00135

Figure Lengend Snippet: IL-6 inhibits TIMP3 expression through STAT3 activation and TIMP3 is upregulated in the cisplatin sensitive group. (A) Western blotting analysis of TIMP3, p-STAT3, and STAT3 protein levels in extracts from Saos2 and Saos2-lung cells treated with 20 ng/mL IL-6 for 24 h. (B,C) The results are expressed as the ratio of TIMP3 to GAPDH, and p-STAT3 to STAT3. (D) The expression of TIMP3 was measured in Saos2 and Saos2-lung cells with or without IL-6 treatment via real-time PCR, with GAPDH as a housekeeping gene. (E) The expression of TIMP3 was measured in Saos2 and Saos2-lung cells via western blotting, with GAPDH as an internal reference. (F) Sub-confluent Saos2 and Saos2-lung cells were grown in serum-free DMEM for 24 h, and the IL-6 level in the supernatant was detected using an ELISA. (G) Immunohistochemical analysis of TIMP3 was carried out in the tumor samples from different patients. Data are presented as the means ± SD. ∗∗ P < 0.01, ∗ P < 0.05. All data were obtained from at least three independent experiments.

Article Snippet: For TIMP3 overexpression, Saos2-lung, MG63, and U2OS cells were transfected with green fluorescent protein (GFP)-labeled-lentiviral vectors (GeneChem, Shanghai, China) expressing TIMP3 or a vector with a scrambled control sequence.

Techniques: Expressing, Activation Assay, Western Blot, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Immunohistochemical staining

Journal: Frontiers in Genetics

Article Title: TIMP3 Overexpression Improves the Sensitivity of Osteosarcoma to Cisplatin by Reducing IL-6 Production

doi: 10.3389/fgene.2018.00135

Figure Lengend Snippet: IL-6 and p-STAT3 expression after the overexpression or knockdown of TIMP3 in Saos2-lung cells. (A) The identification of TIMP3 overexpression and knockdown in Saos2-lung cells. (B) The results are expressed as the ratio of the expression of the target gene to that of GAPDH . (C) The expression levels of p-STAT3 and STAT3 were measured in transfected-Saos2-lung cells treated with or without cisplatin via western blotting, with GAPDH was an internal reference. The results are expressed as the ratio of p-STAT3 to STAT3. (D) IL6 gene expression in transfected-Saos2-lung cells without cisplatin treatment. (E) IL6 gene expression in transfected-Saos2-lung cells with cisplatin treatment. (F) The expression level of TIMP3, p-STAT3, and STAT3 was measured in STAT3 knockdown Saos2-lung cells with or without IL-6. Data are presented as the means ± SD. ∗∗∗ P < 0.001, ∗∗ P < 0.01. All data were obtained from at least three independent experiments.

Article Snippet: For TIMP3 overexpression, Saos2-lung, MG63, and U2OS cells were transfected with green fluorescent protein (GFP)-labeled-lentiviral vectors (GeneChem, Shanghai, China) expressing TIMP3 or a vector with a scrambled control sequence.

Techniques: Expressing, Over Expression, Knockdown, Transfection, Western Blot, Gene Expression

Journal: Frontiers in Genetics

Article Title: TIMP3 Overexpression Improves the Sensitivity of Osteosarcoma to Cisplatin by Reducing IL-6 Production

doi: 10.3389/fgene.2018.00135

Figure Lengend Snippet: Cell proliferation and cisplatin sensitivity after the overexpression or knockdown of TIMP3 in Saos2-lung cells. (A–D) Cisplatin sensitivity of transfected-Saos2-lung cells at 24, 48, 72, and 96 h. (E) Cell proliferation in transfected-Saos2-lung cells without cisplatin. (F) Flow cytometry assessment of apoptosis in transfected-Saos2-lung cells without or with cisplatin treatment. (G) Apoptosis rates of transfected-Saos2-lung cells with or without cisplatin treatment. (H) Hoechst staining of transfected-Saos2-lung after cisplatin treatment for 24 h. (I) The quantification of hoechst staining. The results are expressed as the ratio of p-Akt to t-Akt. Data are presented as the means ± SD. ∗ P < 0.05 between the Saos2-lung and siRNA groups, # P < 0.05 between the Saos2-scramble and Saos2-lung-TIMP3 groups, separately, in a cell proliferation assay. ∗∗∗ P < 0.001, ∗∗ P < 0.01, ∗ P < 0.05. All data were obtained from at least three independent experiments.

Article Snippet: For TIMP3 overexpression, Saos2-lung, MG63, and U2OS cells were transfected with green fluorescent protein (GFP)-labeled-lentiviral vectors (GeneChem, Shanghai, China) expressing TIMP3 or a vector with a scrambled control sequence.

Techniques: Over Expression, Knockdown, Transfection, Flow Cytometry, Staining, Proliferation Assay

Journal: Frontiers in Genetics

Article Title: TIMP3 Overexpression Improves the Sensitivity of Osteosarcoma to Cisplatin by Reducing IL-6 Production

doi: 10.3389/fgene.2018.00135

Figure Lengend Snippet: Apoptosis-related gene expression in Saos2-lung cells following overexpression or knockdown of TIMP3. (A) The expression levels of Bcl-2, Bax, p-Akt, cleaved caspase-3, and cleaved caspase-9 were measured using western blotting in transfected-Saos2-lung cells, with or without cisplatin. (B,C) The results are expressed as the ratio of Bcl-2 to GAPDH, Bax to GAPDH, p-Akt to t-AKT, cleaved caspase-3 to caspase-3, and cleaved caspase-9 to caspase-9. Data are presented as the means ± SD. ∗∗∗ P < 0.001, ∗∗ P < 0.01, ∗ P < 0.05. All data were obtained from at least three independent experiments.

Article Snippet: For TIMP3 overexpression, Saos2-lung, MG63, and U2OS cells were transfected with green fluorescent protein (GFP)-labeled-lentiviral vectors (GeneChem, Shanghai, China) expressing TIMP3 or a vector with a scrambled control sequence.

Techniques: Gene Expression, Over Expression, Knockdown, Expressing, Western Blot, Transfection

Journal: Frontiers in Genetics

Article Title: TIMP3 Overexpression Improves the Sensitivity of Osteosarcoma to Cisplatin by Reducing IL-6 Production

doi: 10.3389/fgene.2018.00135

Figure Lengend Snippet: Cisplatin sensitivity of Saos2-lung cells after TIMP3 overexpression in vivo . (A–C) Cisplatin or saline was used to treat nude mice bearing osteosarcoma cells. Saos2-lung cells with and without TIMP3 overexpression were labeled with luciferase and implanted in the tibia. The tumor volume was evaluated via in vivo bioluminescence assay at 1–5 weeks after cisplatin or saline treatment. (D,E) The mice were sacrificed, and the tumor volume was measured and calculated during the 5th week after treatment. The data are presented as the means ± SD. ∗∗∗ P < 0.001, ∗∗ P < 0.01, ∗ P < 0.05. Each group contained five animals.

Article Snippet: For TIMP3 overexpression, Saos2-lung, MG63, and U2OS cells were transfected with green fluorescent protein (GFP)-labeled-lentiviral vectors (GeneChem, Shanghai, China) expressing TIMP3 or a vector with a scrambled control sequence.

Techniques: Over Expression, In Vivo, Saline, Labeling, Luciferase, ATP Bioluminescent Assay

Journal: Frontiers in Genetics

Article Title: TIMP3 Overexpression Improves the Sensitivity of Osteosarcoma to Cisplatin by Reducing IL-6 Production

doi: 10.3389/fgene.2018.00135

Figure Lengend Snippet: Immunohistochemical analysis of tumor samples with and without TIMP3 overexpression. (A) The expression profiles of TIMP3, cleaved caspase-3, p-Akt, PCNA, and IL-6 were evaluated using immunohistochemistry. (B,C) Quantification of positive staining was performed using Image-Pro Plus 6.0 software. The results are expressed in terms of the mean density of positive staining. (D) Apoptotic cells were detected by TUNEL staining in tissue sections. (E) Semi-quantitative data from the analysis of apoptotic cells. The data are presented as the means ± SD. ∗∗∗ P < 0.001, ∗∗ P < 0.01, ∗ P < 0.05. Each group contained five animals.

Article Snippet: For TIMP3 overexpression, Saos2-lung, MG63, and U2OS cells were transfected with green fluorescent protein (GFP)-labeled-lentiviral vectors (GeneChem, Shanghai, China) expressing TIMP3 or a vector with a scrambled control sequence.

Techniques: Immunohistochemical staining, Over Expression, Expressing, Immunohistochemistry, Staining, Software, TUNEL Assay

Journal: Frontiers in Genetics

Article Title: TIMP3 Overexpression Improves the Sensitivity of Osteosarcoma to Cisplatin by Reducing IL-6 Production

doi: 10.3389/fgene.2018.00135

Figure Lengend Snippet: A schematic diagram of the proposed mechanism of cisplatin sensitivity enhancement by TIMP3 overexpression. According to the results of the current study, IL-6 inhibited TIMP3 expression through STAT3 activation, whereas conversely; TIMP3 suppressed the production of IL-6. TIMP3 also facilitates the expression of cleaved caspase-3, while inhibiting the production of p-Akt, which results in the inhibition of proliferation and the overall potentiation of apoptosis. Taken together, these events contribute to enhancing the sensitivity of osteosarcoma to cisplatin.

Article Snippet: For TIMP3 overexpression, Saos2-lung, MG63, and U2OS cells were transfected with green fluorescent protein (GFP)-labeled-lentiviral vectors (GeneChem, Shanghai, China) expressing TIMP3 or a vector with a scrambled control sequence.

Techniques: Over Expression, Expressing, Activation Assay, Inhibition

Journal: Oncotarget

Article Title: STAT3-mediated activation of miR-21 is involved in down-regulation of TIMP3 and neovascularization in the ischemic retina

doi: 10.18632/oncotarget.21592

Figure Lengend Snippet: (A) QPCR of TIMP3 mRNA expression in control untreated HREC and cells transfected with STAT3-specific siRNA (50 nM final concentration) or negative control siRNA ( NC ) and exposed to hypoxia (pO 2 =2%) for 12 hours. (B) Western blot showing expression of TIMP3 in HREC transfected with STAT3-specific siRNA or NC siRNA. Blots were subjected to densitometry analysis and the obtained data were analyzed for statistical significance. (C) QPCR of TIMP3 mRNA expression in HREC transfected with a.miR-21 (50 nM final concentration) and s.amiR and exposed to 2%O 2 hypoxia for 12 hours. (D) Western blot showing expression of TIMP3 in HREC transfected with a.miR-21 and s.amiR . Blots were subjected to densitometry analysis and the obtained data were analyzed for statistical significance. Values are mean ± SD of four separate experiments.

Article Snippet: Actin antibody was used as an internal control for TIMP3 expression (1:1000; Santa-Cruz Biotech).

Techniques: Expressing, Control, Transfection, Concentration Assay, Negative Control, Western Blot

Journal: Oncotarget

Article Title: STAT3-mediated activation of miR-21 is involved in down-regulation of TIMP3 and neovascularization in the ischemic retina

doi: 10.18632/oncotarget.21592

Figure Lengend Snippet: (A) QPCR of TIMP3 mRNA expression in control HREC and HREC transfected with miR-21 mimic (50 nM) for 24 and 48 hours or mimic NC . (B) Western blot showing expression of TIMP3 in HREC transfected with miR-21 mimic or mimic NC . Values are mean ± SD of four separate experiments.

Article Snippet: Actin antibody was used as an internal control for TIMP3 expression (1:1000; Santa-Cruz Biotech).

Techniques: Expressing, Control, Transfection, Western Blot

Journal: Oncotarget

Article Title: STAT3-mediated activation of miR-21 is involved in down-regulation of TIMP3 and neovascularization in the ischemic retina

doi: 10.18632/oncotarget.21592

Figure Lengend Snippet: (A) QPCR of TIMP3 mRNA expression in retinas of control and OIR mice at P14 and P17. (B) Western blot showing expression of TIMP3 in retinas of OIR and age-matched control mice. Blots were subjected to densitometry analysis and the obtained data were analyzed for statistical significance. Values are mean ± SD (n=5 retinas per group).

Article Snippet: Actin antibody was used as an internal control for TIMP3 expression (1:1000; Santa-Cruz Biotech).

Techniques: Expressing, Control, Western Blot

Journal: Oncotarget

Article Title: STAT3-mediated activation of miR-21 is involved in down-regulation of TIMP3 and neovascularization in the ischemic retina

doi: 10.18632/oncotarget.21592

Figure Lengend Snippet: (A) MiR-21 expression in retinas of control mice and OIR mice at P14 injected with miR-21 inhibitor ( a.miR-21 ) or NC LNA. (B) QPCR of TIMP3 mRNA expression in retinas of control mice and OIR mice at P14 injected with a. miR-21 or NC LNA. (C) Western blot showing expression of TIMP3 in retinas of control mice and OIR mice at P14 injected with a.miR-21 or NC LNA. Blots were subjected to densitometry analysis and the obtained data were analyzed for statistical significance. Values are mean ± SD (n=5 retinas per group).

Article Snippet: Actin antibody was used as an internal control for TIMP3 expression (1:1000; Santa-Cruz Biotech).

Techniques: Expressing, Control, Injection, Western Blot