Journal: Nature Communications

Article Title: MYOD-SKP2 axis boosts tumorigenesis in fusion negative rhabdomyosarcoma by preventing differentiation through p57 Kip2 targeting

doi: 10.1038/s41467-023-44130-0

Figure Lengend Snippet: a Representative western blot ( n = 3 independent experiments) of the indicated proteins on RD and JR1 cells infected with either Scrambled (shSCR), SKP2.1 (shSKP2.1) or SKP2.2 (shSKP2.2) lentiviral shRNA at 72 h post-selection. Vinculin is the loading control. b Representative light microscopy pictures of soft agar colony formation assay on RD and JR1 cells treated as in ( a ) and grown for 2 weeks. Scale bar = 50 μm. c Histogram depicts the quantification of soft agar colony numbers per field. n = 3 (RD) and n = 4 (JR1) independent experiments, data presented as mean values ± SD, one-way ANOVA. d Representative light microscopy pictures of single cell colony formation assay on RD and JR1 cells treated as in ( a ) and grown for 2 weeks. Scale bar = 50 μm. e Histogram depicts the quantification of colony numbers per field. n = 3 independent experiments, data presented as mean values ± SD, one-way ANOVA. f Representative light microscopy pictures of β-Galactosidase staining of RD and JR1 cells transfected with either SCR or SKP2 siRNA. Scale bar = 100 μm. g Histogram depicts the quantification of the percentage of senescent cells per field. n = 3 independent experiments, data presented as mean values ± SD, Student’s two-tailed t -test. h Images of JR1 shSCR, shSKP2.1, and shSKP2.2 tumors explanted from mice post euthanasia at 48 days post-inoculation. i Tumor volume of shSCR ( n = 11), shSKP2.1 ( n = 5) and shSKP2.2 ( n = 6) JR1 xenografts assessed by caliper measurement represented in mm 3 followed for 48 days post-inoculation. Data presented as mean values ± SD, two-way ANOVA. j Tumor weight of shSCR ( n = 11), shSKP2.1 ( n = 5) and shSKP2.2 ( n = 6) JR1 xenografts. Box plots show 25th to 75th quartiles, black bar shows the median, and whiskers go down to the smallest value and up to the largest. One-way ANOVA k Representative images ( n = 3 independent experiments) of H&E, p27 Kip1 , p57 Kip2 , MYOG, MyHC, and Ki67 immunohistochemistry of tumor sections from JR1 xenografts expressing either shSCR or shSKP2.2. Scale Bars = 100 μm. Source data are provided as a Source Data file.

Article Snippet: RD and JR1 cells were infected with lentiviral Lenti-ORF clone of CDKN1C (RC209840L3, Origene, Rockville, MD, USA).

Techniques: Western Blot, Infection, shRNA, Selection, Control, Light Microscopy, Soft Agar Assay, Colony Assay, Staining, Transfection, Two Tailed Test, Immunohistochemistry, Expressing

Journal: European Journal of Medical Research

Article Title: FBXO42 promotes hepatocellular carcinoma progression via mediating p57Kip2 ubiquitination and degradation

doi: 10.1186/s40001-025-03050-z

Figure Lengend Snippet: FBXO42 interacted with p57Kip2 and promoted the ubiquitination and degradation of p57Kip2. A Prediction of FBXO42-interacting molecules via Ubibrowser. B Co-IP assays analysis of the interaction between FBXO42 and p57Kip2. C qRT-PCR analysis of the mRNA of CDKN1C in HCC cells. D Correlation between FBXO42 and CDKN1C in HCC based on GEPIA database. E Western blotting analysis of the protein level of p57Kip2 in HCC cells. F HepG2 cells were incubated with or without 10 μM MG132 for 6 h, and the protein levels of p57Kip2 were subsequently assessed via Western blotting analysis. G Relative levels of p57Kip2 protein of HepG2 cells treated with cycloheximide (CHX) for the indicated time periods and were plotted. H Total ubiquitination levels of p57Kip2 were examined in HepG2 cells overexpressing FBXO42 following transfection with corresponding plasmids for 48 h and subsequent treatment with MG132 (10 μM) for 6 h. ns not significant; ** p < 0.01

Article Snippet: The primary antibodies used in this study: FBXO42 (Invitrogen), GAPDH (Proteintech), p57Kip2 (Proteintech), anti-Flag (Proteintech), anti-HA (Proteintech) and YY1 (Proteintech).

Techniques: Ubiquitin Proteomics, Co-Immunoprecipitation Assay, Quantitative RT-PCR, Western Blot, Incubation, Transfection

Journal: European Journal of Medical Research

Article Title: FBXO42 promotes hepatocellular carcinoma progression via mediating p57Kip2 ubiquitination and degradation

doi: 10.1186/s40001-025-03050-z

Figure Lengend Snippet: FBXO42 facilitated the malignant behaviors via p57Kip2 in HCC. A Protein levels of FBXO42 and p57Kip2 in HepG2 cells were evaluated by Western blotting. B – D CCK-8 assays, clone formation assays and EDU staining assays were conducted to evaluate the impact of CDKN1C overexpression on cell proliferation in HepG2 cells with FBXO42 overexpression. E Transwell assays were conducted to assess the impact of CDKN1C overexpression on cell migration in HepG2 cells with FBXO42 overexpression. F – H Representative images, growth cruse and weight of HCC xenografts revealed the effect of CDKN1C overexpression on subcutaneous tumor growth in HepG2 cells overexpressing FBXO42. I, J Representative images from IHC staining for Ki-67 and TUNEL expression in xenograft HCC tumors. Scale bar: 100 μm. * p < 0.05; ** p < 0.01; *** p < 0.001

Article Snippet: The primary antibodies used in this study: FBXO42 (Invitrogen), GAPDH (Proteintech), p57Kip2 (Proteintech), anti-Flag (Proteintech), anti-HA (Proteintech) and YY1 (Proteintech).

Techniques: Western Blot, CCK-8 Assay, Staining, Over Expression, Migration, Immunohistochemistry, TUNEL Assay, Expressing

Journal: European Journal of Medical Research

Article Title: FBXO42 promotes hepatocellular carcinoma progression via mediating p57Kip2 ubiquitination and degradation

doi: 10.1186/s40001-025-03050-z

Figure Lengend Snippet: YY1 upregulated the level of FBXO42 in HCC. A Prediction of potential upstream transcriptional regulators of FBXO42 via hTFtarget, ENCODE and CHEA and TCGA databases. B Impact of CTCF and YY1 on overall survival of HCC patients. C Expression levels of YY1 in tumor tissues and their normal tissues, and expression analysis of YY1 in 50 HCC tissues and their corresponding adjacent normal liver tissues were analyzed through TCGA database. D Correlation between FBXO42 and YY1 was evaluated in HCC utilizing data from the TCGA and GEPIA databases. E – H mRNA and protein level of FBXO42 and YY1 in HepG2 and Hep3B cells. I, J Dual luciferase reporter assays were conducted to verify that YY1 activates FBXO42 transcription in HCC. K Graphical abstract showed YY1-upregulated FBXO42 mediated ubiquitination and degradation of p57Kip2, consequently promoting the malignant behaviors of HCC. *** p < 0.001

Article Snippet: The primary antibodies used in this study: FBXO42 (Invitrogen), GAPDH (Proteintech), p57Kip2 (Proteintech), anti-Flag (Proteintech), anti-HA (Proteintech) and YY1 (Proteintech).

Techniques: Expressing, Luciferase, Ubiquitin Proteomics

Journal: Journal of cellular biochemistry

Article Title: MicroRNA-145-5p suppresses cell proliferation, migration, and invasion in upper tract urothelial carcinoma by targeting 5-aminoimidazole-4-carboxamide ribonucleotide formyltransferase/IMP cyclohydrolase.

doi: 10.1002/jcb.30449

Figure Lengend Snippet: FIGURE 5 RNA‐sequencing analysis of gene expression by 5‐aminoimidazole‐4‐carboxamide ribonucleotide formyltransferase/IMP cyclohydrolase (ATIC) knockdown in BFTC909 and KMPC3 cells. (A) miR‐145‐5p overexpression and ATIC knockdown reduced phospho‐ p70 S6KThr389 protein levels in BFTC909 cells. (B) Heatmap showed the top 32 upregulated and downregulated genes in small interfering RNA (siRNA)‐ATIC and siRNA‐control transfected BFTC909 and KPMC3 cells. Log2 expression values were row‐normalized using Z‐scores. Red and green colors indicate decreased and increased expression, respectively. (C) The network of the top 32 putative genes identified by Ingenuity Pathway Analysis (IPA) (D) IPA canonical pathways analyzed the top 32 upregulated and downregulated genes in siRNA‐ATIC and siRNA‐control transfected UTUC cells. Data indicated that these genes were correlated with some diseases, molecular functions, and physiological system functions. (E) ATIC knockdown reduced the protein levels of FN1, Slug, and cyclin A2 and induced p57 in BFTC909 cells (N = 3). (F) Overexpression of miR‐145‐5p decreased the protein levels of FN1, Slug, cyclin A2, and cyclin B1 and increased the expression of p57 in BFTC909 cells (N = 3). Results are shown as the mean ± standard deviation; *p < 0.05 and **p < 0.01. N.S., not significant.

Article Snippet: See the T erm s and C onditions (https://onlinelibrary.w iley.com /term s-and-conditions) on W iley O nline L ibrary for rules of use; O A articles are governed by the applicable C reative C om m ons L icense Technology), fibronectin 1 (FN1) (GTX112794, GeneTex), Slug (NBP2‐52570, Novus), cyclin A2 (#4656, Cell Signaling Technology), cyclin B1 (#4138, Cell Signaling Technology), p57 (NBP1‐89917, Novus), and interferon‐ induced transmembrane 1 (IFITM1) (NBP1‐89345, Novus).

Techniques: RNA Sequencing, Gene Expression, Knockdown, Over Expression, Small Interfering RNA, Control, Transfection, Expressing, Standard Deviation

Journal: Journal of cellular biochemistry

Article Title: MicroRNA-145-5p suppresses cell proliferation, migration, and invasion in upper tract urothelial carcinoma by targeting 5-aminoimidazole-4-carboxamide ribonucleotide formyltransferase/IMP cyclohydrolase.

doi: 10.1002/jcb.30449

Figure Lengend Snippet: FIGURE 6 5‐aminoimidazole‐4‐carboxamide ribonucleotide formyltransferase/IMP cyclohydrolase (ATIC) reversed the inhibitory effects of miR‐145‐5p in upper tract urothelial carcinoma (UTUC) cells. (A) miR‐145‐5p inhibited proliferation in BFTC909 and UM‐UC‐14 cells but was restored by ATIC overexpression (N = 3). (B) miR‐145‐5p repressed the migration abilities of BFTC909 and UM‐UC‐14 cells but they were restored by ATIC overexpression (N = 3). (C) miR‐145‐5p suppressed the invasion abilities of BFTC909, UM‐UC‐14, and KMPC3 cells but they were restored by ATIC overexpression (N = 3). (D) Western blot analysis revealed that ATIC, FN1, Slug, cyclin A2, cyclin B1, P57, and IFITM1 levels were restored after the cotransfection of miR‐145‐5p mimics and CMV‐ATIC compared with the levels in BFTC909 cells transfected with miR‐145‐5p alone with α‐tubulin as a reference. (E) Quantification of the protein levels of ATIC, FN1, Slug, cyclin A2, cyclin B1, P57, and IFITM1 from (D) (N = 3). (F) Analysis of miR‐145‐5p, ATIC, IFITM1, CDKN1C, FN1, SNAI2, CCNA2, and CCNB1 expression in UTUC tissues compared with that in adjacent normal tissues using GSE159824 database (N = 2). Data are shown as the mean ± standard deviation; *p < 0.05, **p < 0.01, and ***p < 0.001.

Article Snippet: See the T erm s and C onditions (https://onlinelibrary.w iley.com /term s-and-conditions) on W iley O nline L ibrary for rules of use; O A articles are governed by the applicable C reative C om m ons L icense Technology), fibronectin 1 (FN1) (GTX112794, GeneTex), Slug (NBP2‐52570, Novus), cyclin A2 (#4656, Cell Signaling Technology), cyclin B1 (#4138, Cell Signaling Technology), p57 (NBP1‐89917, Novus), and interferon‐ induced transmembrane 1 (IFITM1) (NBP1‐89345, Novus).

Techniques: Over Expression, Migration, Western Blot, Cotransfection, Transfection, Expressing, Standard Deviation

Journal: Cell Death & Disease

Article Title: Increased expression of long noncoding RNA TUG1 predicts a poor prognosis of gastric cancer and regulates cell proliferation by epigenetically silencing of p57

doi: 10.1038/cddis.2015.356

Figure Lengend Snippet: TUG1 is associated with PRC2 in GC. ( a ) The expression of p15, p16, p21, p27 and p57 was determined after knockdown of TUG1 using qRT-PCR. ( b ) TUG1 nuclear localization, as identified using qRT-PCR in fractionated BGC-823 and AGS cells. After nuclear and cytosolic separation, RNA expression levels were measured by qRT-PCR. GAPDH was used as a cytosolic marker, and U6 was used as a nuclear marker. ( c ) RIP experiments were performed, and the coprecipitated RNA was subjected to qRT-PCR for TUG1. The fold enrichment of TUG1 in RIPs is relative to its matching IgG control RIP. * P <0.05, ** P <0.01

Article Snippet: The p57 and p21 siRNAs were purchased from Santa Cruz (Dallas, TX, USA) (cat. no. sc-35751 and sc-29427).

Techniques: Expressing, Knockdown, Quantitative RT-PCR, RNA Expression, Marker, Control

Journal: Cell Death & Disease

Article Title: Increased expression of long noncoding RNA TUG1 predicts a poor prognosis of gastric cancer and regulates cell proliferation by epigenetically silencing of p57

doi: 10.1038/cddis.2015.356

Figure Lengend Snippet: TUG1 is required to target PRC2 occupancy and activity to epigenetically regulate the expression of CKIs, thus regulating GC cell cycle and proliferation. ( a ) The expression of p15, p16, p21, p27 and p57 in BGC-823 and AGS cells, after knockdown of EZH2 and SUZ12. ( b ) ChIP-qPCR of H3K27me3 and EZH2 of the promoter region of the p15, p16, p21, p27 and p57 locus after siRNA treatment targeting si-NC or si-TUG1 in AGS cells. Antibody enrichment was quantified relative to the amount of input DNA. Antibody directed against IgG was used as a negative control. ( c ) The expression of EZH2 and SUZ12 in BGC-823 and AGS cells, after knockdown of TUG1. * P <0.05, ** P <0.01

Article Snippet: The p57 and p21 siRNAs were purchased from Santa Cruz (Dallas, TX, USA) (cat. no. sc-35751 and sc-29427).

Techniques: Activity Assay, Expressing, Knockdown, ChIP-qPCR, Negative Control

Journal: Cell Death & Disease

Article Title: Increased expression of long noncoding RNA TUG1 predicts a poor prognosis of gastric cancer and regulates cell proliferation by epigenetically silencing of p57

doi: 10.1038/cddis.2015.356

Figure Lengend Snippet: The role of p57 in GC. ( a ) Immunohistochemistry was used to detect the expression of EZH2 protein in 30 pairs of GC with corresponding non-tumor tissues. Bar, 100 μ m. The immunoreactivity of EZH2 protein in GC tissues showed a statistically significant positive correlation with the relative level of TUG1 expression. ( b ) AGS and BGC-823 cells transfected with si-NC/si-EZH2. Forty-eight hours after transfection, the cells were analyzed using flow cytometry. ( c ) As determined by qRT-PCR assays, the level of p57 was downregulated in 30 pairs of GC tissues. Western blot assays detected the expression of p57 after transfection. AGS cells were transfected with Vector/p57. Forty-eight hours after transfection, the cells were analyzed using MTT assays and flow cytometry. AGS cells were transfected with si-NC/si-TUG1/si-TUG1+vector and transfected with si-TUG1 followed by transfection with pcDNA-p57. After transfection, the cells were stained and analyzed using flow cytometry. MTT analysis of cell proliferation by co-transfection (si-NC, si-TUG1 1#, si-TUG1 1#+si-NC, si-TUG1 1#+si-p57, si-TUG1 1#+si-p21). * P <0.05, ** P <0.01

Article Snippet: The p57 and p21 siRNAs were purchased from Santa Cruz (Dallas, TX, USA) (cat. no. sc-35751 and sc-29427).

Techniques: Immunohistochemistry, Expressing, Transfection, Flow Cytometry, Quantitative RT-PCR, Western Blot, Plasmid Preparation, Staining, MTT Cell Proliferation, Cotransfection