anti uck2  (Proteintech)

Journal: EMBO Molecular Medicine

Article Title: De novo pyrimidine synthesis is a collateral metabolic vulnerability in NF2 -deficient mesothelioma

doi: 10.1038/s44321-025-00278-4

Figure Lengend Snippet: ( A ) Cell viability of H2452 NF2 wild-type (sgCtrl) and NF2 knockout (sg NF2 -1, sg NF2 -2) cells treated with various doses of the IMPDH inhibitor for 96 h. Representative data from three independent experiments are shown. The data are presented as the mean ± SD. Two-way ANOVA with multiple comparisons was used for statistical analysis. ( B ) Cell viability of H2452 NF2 wild-type (sgCtrl) and NF2 knockout (sg NF2 -1, sg NF2 -2) cells cultured with the indicated concentrations of glucose. The data are presented as the mean ± SD ( n = 3). Two-way ANOVA with multiple comparisons was used for statistical analysis. ( C–E ) Cell viability of H2452 NF2 wild-type (sgCtrl) and NF2 knockout (sg NF2 -1, sg NF2 -2) cells treated with the indicated doses of the UCK2 inhibitor for 96 h at 10 mM ( C ), 5 mM ( D ) and 2.5 mM ( E ) glucose. Representative data from three independent experiments are shown. The data are presented as the mean ± SD. Two-way ANOVA with multiple comparisons was used for statistical analysis. ( F–H ) Cell viability of H2452 NF2 wild-type (sgCtrl) and NF2 knockout (sg NF2 -1, sg NF2 -2) cells treated with the indicated doses of the DHODH inhibitor for 96 h at 10 mM ( F ), 5 mM ( G ) and 2.5 mM ( H ) glucose. Representative data from three independent experiments were shown. The data are presented as the mean ± SD. Two-way ANOVA with multiple comparisons was used for statistical analysis. ( I ) The differences in LFQ intensities from 480 label-free proteomes of the indicated markers between the H2452 sg NF2 -1 and sgCtrl groups. The data are presented as the mean ± SD ( n = 3). A two-tailed unpaired t test was used for statistical analysis. ( J , K ) The mRNA expression of UPP1 ( J ) and cell viability ( K ) of the indicated cell populations transfected with small interfering RNA (siRNA) targeting the negative control or UPP1 for 72 h. The data are presented as the mean ± SD ( n = 3). Two-way ANOVA with multiple comparisons was used for statistical analysis.

Article Snippet: UCK2 inhibitor , MedChemExpress , HY-148394.

Techniques: Knock-Out, Cell Culture, Two Tailed Test, Expressing, Transfection, Small Interfering RNA, Negative Control

Journal: Translational Cancer Research

Article Title: Uridine-cytidine kinase 2 as a novel potential prognostic and therapeutic biomarker in gastric cancer

doi: 10.21037/tcr-2025-1165

Figure Lengend Snippet: High UCK2 expression is closely correlated with more advanced TNM staging and a worse clinical prognosis. (A-C) UCK2 was differently expressed in the GC tissues compared to the non-tumor tissues in TCGA (A), TCGA with GTEx (B), and GEO (C) datasets. (D) A survival analysis was performed using the GEO datasets. (E) IHC staining was performed to detect UCK2 expression. (F) IHC grades were examined. (G) A Kaplan-Meier analysis was conducted based on different UCK2 expression levels to assess the OS of the GC patients. (H) Forest plot showing the independent predictors of prognosis through a Cox regression model. Data are expressed as the mean ± standard deviation unless otherwise stated. * P<0.05, *** P<0.001. CI, confidence interval; GC, gastric cancer; GEO, Gene Expression Omnibus; GTEx, Genotype-Tissue Expression; HR, hazard ratio; IHC, immunohistochemistry; OS, overall survival; pN, pathological N; TCGA, The Cancer Genome Atlas; TNM, tumor-node-metastasis; UCK2, uridine-cytidine kinase 2.

Article Snippet: Briefly, deparaffinized paraffin-embedded tissue sections underwent 30-minute heating for antigen retrieval, followed by incubation overnight at 4 °C with anti-UCK2 antibody (1:200; Cat. No. 10511-1-AP, Proteintech, Wuhan, China).

Techniques: Expressing, Immunohistochemistry, Standard Deviation, Gene Expression

Journal: Translational Cancer Research

Article Title: Uridine-cytidine kinase 2 as a novel potential prognostic and therapeutic biomarker in gastric cancer

doi: 10.21037/tcr-2025-1165

Figure Lengend Snippet: UCK2 expression levels in GC cell lines and the establishment of silenced UCK2 cell lines. (A,B) The expression levels of UCK2 in the GC cell lines were detected by qPCR (A) and western blot (B). (C,D) Stable cell lines expressing shRNA plasmids were constructed, and their characterization was confirmed by qPCR (C) and western blot (D). (E) Stable UCK2-knocked-out gastric cell lines were also established using the CRSPR/Cas9 techniques. UCK2 expression was examined by western blot. Data are expressed as the mean ± standard deviation unless otherwise stated. ***, P<0.001. GC, gastric cancer; NC, negative control; qPCR, quantitative polymerase chain reaction; shRNA, short hairpin RNA; UCK2, uridine-cytidine kinase 2.

Article Snippet: Briefly, deparaffinized paraffin-embedded tissue sections underwent 30-minute heating for antigen retrieval, followed by incubation overnight at 4 °C with anti-UCK2 antibody (1:200; Cat. No. 10511-1-AP, Proteintech, Wuhan, China).

Techniques: Expressing, Western Blot, Stable Transfection, shRNA, Construct, Standard Deviation, Negative Control, Real-time Polymerase Chain Reaction

Journal: Translational Cancer Research

Article Title: Uridine-cytidine kinase 2 as a novel potential prognostic and therapeutic biomarker in gastric cancer

doi: 10.21037/tcr-2025-1165

Figure Lengend Snippet: The down-regulation of UCK2 attenuated GC cell viability and proliferation. (A,B) The knock-down of UCK2 impaired GC cell viability and proliferation in the AGS and HCG27 cells as shown by CCK-8 (A) and colony formation (crystal violet staining; magnification, XXXXXX) (B) assays. (C,D) The knock-out of UCK2 inhibited the survival and proliferation ability of the GC cells as demonstrated by CCK-8 (C) and colony formation (crystal violet staining; magnification, XXX) (D) assays. (E) AGS cells with control or sgRNA plasmids were subcutaneously injected into the male Balb/c nude mice to establish an in vivo xenograft model (n=6). (F-H) Image showing tumor volumes (F), tumor growth curves (G), and tumor weight (H) for each group. Data are expressed as the mean ± standard deviation unless otherwise stated. **, P<0.01; ***, P<0.001. GC, gastric cancer; NC, negative control; OD, optical density; SEM, standard error of the mean; sgRNA, single guide RNA; shRNA, short hairpin RNA; UCK2, uridine-cytidine kinase 2.

Article Snippet: Briefly, deparaffinized paraffin-embedded tissue sections underwent 30-minute heating for antigen retrieval, followed by incubation overnight at 4 °C with anti-UCK2 antibody (1:200; Cat. No. 10511-1-AP, Proteintech, Wuhan, China).

Techniques: Knockdown, CCK-8 Assay, Staining, Knock-Out, Control, Injection, In Vivo, Standard Deviation, Negative Control, shRNA

Journal: Translational Cancer Research

Article Title: Uridine-cytidine kinase 2 as a novel potential prognostic and therapeutic biomarker in gastric cancer

doi: 10.21037/tcr-2025-1165

Figure Lengend Snippet: UCK2 enhances GC cell motility and metastasis. (A,B) Transwell assays showed that the knock-down of UCK2 inhibited the migration (A) and invasion (B) ability of the GC cells. Crystal violet staining; scale bar: XXX. (C,D) The knock-out of UCK2 significantly impaired the motility of GC cells as demonstrated by the migration assays (C) and invasion (D) assays. Crystal violet staining; scale bar: XXX. (E) A tumor peritoneal metastasis mouse model was established by the intraperitoneal injection of AGS cells with control or sgRNA plasmids into male Balb/c nude mice (n=3). The abdominal tumor metastatic sites (red arrows) are presented. Data are expressed as the mean ± standard deviation unless otherwise stated. **, P<0.01; ***, P<0.001. GC, gastric cancer; NC, negative control; sgRNA, single guide RNA; shRNA, short hairpin RNA; UCK2, uridine-cytidine kinase 2.

Article Snippet: Briefly, deparaffinized paraffin-embedded tissue sections underwent 30-minute heating for antigen retrieval, followed by incubation overnight at 4 °C with anti-UCK2 antibody (1:200; Cat. No. 10511-1-AP, Proteintech, Wuhan, China).

Techniques: Knockdown, Migration, Staining, Knock-Out, Injection, Control, Standard Deviation, Negative Control, shRNA

Journal: Translational Cancer Research

Article Title: Uridine-cytidine kinase 2 as a novel potential prognostic and therapeutic biomarker in gastric cancer

doi: 10.21037/tcr-2025-1165

Figure Lengend Snippet: UCK2 altered the gene expression profile and regulated the cell cycle in GC. (A) Patients in TCGA database were divided into high- and low-expression groups based on their UCK2 expression levels. Volcano plots were the generated from the mRNA-sequencing analysis of the two groups. (B,C) KEGG (B) and GO (C) enrichment analyses were performed on all the DEGs in TCGA dataset. (D,E) RNA sequencing was used to examine the mRNA profile of the silenced-UCK2 AGS cells and the non-silenced control AGS cells, and a total of 1,014 DEGs were identified (|log 2 FC| ≥1 and P<0.05). After conducting the mRNA-sequencing analysis, we generated volcano plots (D) and conducted the KEGG enrichment analysis (E) based on the resulting data. (F) Western blotting was performed to analyze the roles of UCK2 in AKT, cyclin D1, and cyclin E1 expression. (G) AKT kinase inhibitor (HY-10249A from MCE, HY) was used to treat the GC cells. AKT kinase inhibitor (HY) blocked the effect of UCK2 knock-down/knock-out on the p-AKT, cyclin D1, and cyclin E1 expression. The red box marks the signaling pathway and its downstream signals, which were the key focus of the study. DEGs, differentially expressed genes; FC, fold change; GC, gastric cancer; GO, Gene Ontology; KEGG, Kyoto Encyclopedia of Genes and Genomes; mRNA, messenger RNA; NC, negative control; TCGA, The Cancer Genome Atlas; UCK2, uridine-cytidine kinase 2.

Article Snippet: Briefly, deparaffinized paraffin-embedded tissue sections underwent 30-minute heating for antigen retrieval, followed by incubation overnight at 4 °C with anti-UCK2 antibody (1:200; Cat. No. 10511-1-AP, Proteintech, Wuhan, China).

Techniques: Gene Expression, Expressing, Generated, Sequencing, RNA Sequencing, Control, Western Blot, Knockdown, Knock-Out, Negative Control