Journal: Cancers

Article Title: Integrin α2 and β1 Cross-Communication with mTOR/AKT and the CDK-Cyclin Axis in Hepatocellular Carcinoma Cells

doi: 10.3390/cancers14102430

Figure Lengend Snippet: IGF1R expression level in HepG2, Huh7, and Hep3B cell lines. MFU depicts mean fluorescence units. * indicates significant difference to Ms IgG1 ( n = 3). Flow cytometry curves are representative for one of the three experiments.

Article Snippet: The following monoclonal antibodies were applied: Anti-CDK1 (IgG1, clone 1); anti-pCDK1/Cdc2 (pY15; IgG1, clone 44/Cdk1/Cdc2); anti-CDK2 (IgG2a, clone 55); anti-Cyclin A (IgG1, clone 25); anti-Cyclin B (IgG1, clone 18); anti-p19 (IgG1, clone 52); Kip1/p27 (IgG1, clone 57) (all obtained from BD Biosciences), and anti-pCDK2 (Thr160; Cell Signaling, Danvers, MA, USA).

Techniques: Expressing, Fluorescence, Flow Cytometry

Journal: Cancers

Article Title: Integrin α2 and β1 Cross-Communication with mTOR/AKT and the CDK-Cyclin Axis in Hepatocellular Carcinoma Cells

doi: 10.3390/cancers14102430

Figure Lengend Snippet: ( A ) Protein profile of cell cycle regulating proteins (AKT, mTOR, Rictor, Raptor, all total and phosphorylated, “p”), CDK1 and 2 (total and phosphorylated, “p”), Cyclin A and B, FAK, pFAK and ILK. HepG2 cells were either stimulated with IGF or stimulated with IGF and treated with an integrin α2 or β1 specific siRNA (scr = scrambled siRNA). Following siRNA transfection protein data of integrin α2 and β1 are shown on the right side of A. Controls ( C ) received cell culture medium alone. One representative experiment of three is shown; ( B ) Quantification of the protein intensity in IGF1 treated versus non-treated HepG2 cells by the pixel density analysis, all related to the unstimulated 100% controls. * significant difference to controls; ( C , D ): Pixel density analysis of the protein level in HepG2 cells following α2 ( C ) or β1 ( D ) siRNA knock-down. Values are given in percentage, related to the 100% control. * significant difference to controls.

Article Snippet: The following monoclonal antibodies were applied: Anti-CDK1 (IgG1, clone 1); anti-pCDK1/Cdc2 (pY15; IgG1, clone 44/Cdk1/Cdc2); anti-CDK2 (IgG2a, clone 55); anti-Cyclin A (IgG1, clone 25); anti-Cyclin B (IgG1, clone 18); anti-p19 (IgG1, clone 52); Kip1/p27 (IgG1, clone 57) (all obtained from BD Biosciences), and anti-pCDK2 (Thr160; Cell Signaling, Danvers, MA, USA).

Techniques: Transfection, Cell Culture

Journal: Cancers

Article Title: Integrin α2 and β1 Cross-Communication with mTOR/AKT and the CDK-Cyclin Axis in Hepatocellular Carcinoma Cells

doi: 10.3390/cancers14102430

Figure Lengend Snippet: Integrin β1 interacts with AKT. IP was performed wherein integrin β1 was pulled down using mouse anti Human CD29 (Clone:18/CD29). Following SDS-PAGE electrophoresis and WB, the separated proteins were probed with antibodies against β1, AKT, and pAKT. Lysate from cells treated with IGF1 (+IGF) demonstrated strong interaction of pAKT with integrin β1. Lysate from cells treated with siRNA β1 showed knockdown efficiency of β1 protein. Mouse IgG was used as a negative con-trol. A total of 20 µg of cell lysate was used as input. IP, immunoprecipitation; WB, Western blotting.

Article Snippet: The following monoclonal antibodies were applied: Anti-CDK1 (IgG1, clone 1); anti-pCDK1/Cdc2 (pY15; IgG1, clone 44/Cdk1/Cdc2); anti-CDK2 (IgG2a, clone 55); anti-Cyclin A (IgG1, clone 25); anti-Cyclin B (IgG1, clone 18); anti-p19 (IgG1, clone 52); Kip1/p27 (IgG1, clone 57) (all obtained from BD Biosciences), and anti-pCDK2 (Thr160; Cell Signaling, Danvers, MA, USA).

Techniques: SDS Page, Electrophoresis, Immunoprecipitation, Western Blot

Journal: Scientific Reports

Article Title: Novel therapeutic strategy for melanoma based on albendazole and the CDK4/6 inhibitor palbociclib

doi: 10.1038/s41598-022-09592-0

Figure Lengend Snippet: ABZ arrested the melanoma cell cycle at the G2/M phase and induced cell apoptosis. ( A ) Summary of the distribution of differentially expressed genes identified from RNA sequencing data generated from A375 cells with or without ABZ treatment. The average log CPM (count per million) represented the gene expression level. The color represents differentially expressed genes (fold change > 1.5, FDR < 0.05; red: upregulation, green: downregulation, gray: nonsignificant change). B-C. Top enriched GO ( B ) and KEGG ( C ) biological process pathways for genes upregulated or downregulated in A375 cells treated with ABZ. D-E. Representative image ( D ) and quantification ( E ) of flow cytometry analysis of the cell cycle in A375 and SK-MEL-28 cells after treatment with the indicated doses of ABZ. Bar graphs show the quantified results of three independent experiments and are shown as the means ± SD, NS p > 0.05, * p < 0.05, ** p < 0.01, *** p < 0.001. ( F ) Representative image (full-length gels were presented in Supplementary Figure B) of western blot analysis of CDK1, Cyclin B1, p53, p-RB1 and RB1 in A375 and SK-MEL-28 cells after treatment with the indicated doses of ABZ. GAPDH was used as the loading control. Representative image ( G ) and quantification ( H ) of flow cytometry analysis of apoptosis in A375 and SK-MEL-28 cells after treatment with the indicated doses of ABZ. Bar graphs show the quantified results of three independent experiments and are shown as the means ± SD, *** p < 0.001.

Article Snippet: For western blot analysis, the following antibody dilutions were used: CDK1 (Santa Cruz, sc-54), Cyclin B1 (Santa Cruz, sc-245), p53 (Santa Cruz, sc-126), p-RB1 (Cell Signaling Technology, 8516), RB1 (Cell Signaling Technology, 9313), Cleaved Caspase-3 (Cell Signaling Technology, 9661), CDK4 (Santa Cruz, sc-238969), Cyclin D (Santa Cruz, sc-8396), phospho Histone H3 (Hunan AiFang biological, Co., Ltd, AF00582) and GAPDH (Santa Cruz, sc-47724).

Techniques: RNA Sequencing, Generated, Gene Expression, Flow Cytometry, Western Blot, Control

Journal: Scientific Reports

Article Title: Novel therapeutic strategy for melanoma based on albendazole and the CDK4/6 inhibitor palbociclib

doi: 10.1038/s41598-022-09592-0

Figure Lengend Snippet: Synergistic effect of ABZ and palbociclib in the treatment of a melanoma xenograft mouse model. ( A ) A375 cells were injected into mice (n = 4 mice per group) on Day -6, and 50 mg/kg ABZ and 20 mg/kg palbociclib were administered as indicated. ( B ) Tumor volume was measured after ABZ, palbociclib, or ABZ plus palbociclib treatment at the indicated time points. Data represent mean ± SD, *** p < 0.001. ( C ) Representative images of tumors after ABZ, palbociclib, or ABZ plus palbociclib treatment at the end of the experiment in the mouse model. ( D ) Tumor weight was measured on Day 19. Data represent mean ± SD, ** p < 0.01, *** p < 0.001. ( E ) Mouse weight was measured at the indicated time points. F-G. Representative images ( F ) and quantification ( G ) of immunohistochemistry for Ki67, CDK1 and p-RB1 in tumor sections from mouse xenografted melanoma tumors. Data represent mean ± SD, * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: For western blot analysis, the following antibody dilutions were used: CDK1 (Santa Cruz, sc-54), Cyclin B1 (Santa Cruz, sc-245), p53 (Santa Cruz, sc-126), p-RB1 (Cell Signaling Technology, 8516), RB1 (Cell Signaling Technology, 9313), Cleaved Caspase-3 (Cell Signaling Technology, 9661), CDK4 (Santa Cruz, sc-238969), Cyclin D (Santa Cruz, sc-8396), phospho Histone H3 (Hunan AiFang biological, Co., Ltd, AF00582) and GAPDH (Santa Cruz, sc-47724).

Techniques: Injection, Immunohistochemistry

Journal: Biochemistry and Biophysics Reports

Article Title: miR-21-5p inhibitor enhances the radiosensitivity of human cervical cancer cells via blocking CPEB3-mediated CDK1/cyclin B pathway

doi: 10.1016/j.bbrep.2025.102177

Figure Lengend Snippet: miR-21-5p inhibitor directly targeting CPEB3-mediated suppression of CDK1/cyclin B. (A) Bioinformatics analysis revealed the predicted binding sites between miR-21-5p and CPEB3; (B) Effect of miR-21-5p on mRNA expression level of CPEB3 in human cervical cancer cells. HeLa or SiHa cells were transfected with miR-21-5p for 48 h, the mRNA expression level of CPEB3 was determined by RT-PCR; (C) Luciferase reporter assay was performed to measure the binding activity and key binding sites between CPEB3 and miR-21-5p; (D) Effect of miR-21-5p on the expression of CPEB3-mediated CDK1/cyclin B. HeLa or SiHa cells were treated by miR-21-5p inhibitor for 48 h, the expression of CPEB3, CDK1, Cyclin B and β-actin were determined by immunoblot analysis; ∗∗p < 0.01.

Article Snippet: The information of primary antibodies as follow CDK1 (CST #18048), cyclin B (CST # #12231), β-actin (CST #3700), CPEB3 (Invitrogen #12669-1-AP).

Techniques: Binding Assay, Expressing, Transfection, Reverse Transcription Polymerase Chain Reaction, Luciferase, Reporter Assay, Activity Assay, Western Blot

Journal: Journal of Ovarian Research

Article Title: LncRNA TONSL-AS1 regulates miR-490-3p/CDK1 to affect ovarian epithelial carcinoma cell proliferation

doi: 10.1186/s13048-020-00657-0

Figure Lengend Snippet: Upregulation of CDK1 was observed after the overexpression of TONSL-AS1. The effects of overexpression of TONSL-AS1 and miR-490-3p on the expression of CDK1 mRNA (Fig. 3 a ) and protein (Fig. 3 b ) in OVCAR3 cells were analyzed by qPCR and western blot, respectively. Experiments were repeated 3 times and data were expressed as mean values, *, p < 0.05

Article Snippet: Membranes were blocked in 5% BSA (TBST), followed by incubation with CDK1 (ab131450, Abcam) and GAPDH (ab9485, Abcam) primary antibodies at 4 °C for 18 h, followed by incubation with secondary antibody of HRP Goat Anti-Rabbit (IgG) (ab6721, Abcam).

Techniques: Over Expression, Expressing, Western Blot

Journal: Journal of Ovarian Research

Article Title: LncRNA TONSL-AS1 regulates miR-490-3p/CDK1 to affect ovarian epithelial carcinoma cell proliferation

doi: 10.1186/s13048-020-00657-0

Figure Lengend Snippet: TONSL-AS1 regulated the miR-490-3p/CDK1 axis to promote cell proliferation. CCK-8 assay was performed to analyze the effects of overexpression of TONSL-AS1, miR-490-3p and CDK1 on proliferation of OVCAR3 cells. Experiments were repeated 3 times and data were expressed as mean values, *, p < 0.05

Article Snippet: Membranes were blocked in 5% BSA (TBST), followed by incubation with CDK1 (ab131450, Abcam) and GAPDH (ab9485, Abcam) primary antibodies at 4 °C for 18 h, followed by incubation with secondary antibody of HRP Goat Anti-Rabbit (IgG) (ab6721, Abcam).

Techniques: CCK-8 Assay, Over Expression

Journal: Cancers

Article Title: Molecular Comparison of Imatinib-Naïve and Resistant Gastrointestinal Stromal Tumors: Differentially Expressed microRNAs and mRNAs

doi: 10.3390/cancers11060882

Figure Lengend Snippet: Ingenuity Pathway Analysis indicates the relation between genes differentially expressed between imatinib-naïve (IM-n) and imatinib-resistant (IM-r) gastrointestinal stromal tumor (GIST) samples. The 352 significant differentially expressed genes between IM-r and IM-n GIST samples were used as input for an Ingenuity Pathway Analysis (IPA). The depicted IPA networks illustrate and visualize associations between the genes. ( A ) IPA network highlighting cell cycle related, differentially expressed genes. ( B ) IPA network highlighting CDK1, AURKB, and FOXM1 interactions. Green and red shading indicates relatively low and high expression in the IM-r samples. See for an extended symbol legend.

Article Snippet: The cDNA was used in a PCR reaction using primer/probe combinations from the following Taqman ® gene expression assays (AURKA, assay ID: Hs01582072_m1; AURKB, assay ID: Hs00945858_g1; CCND2, assay ID:Hs00153380_m1; CCNE2, assay ID: Hs00180319_m1; CDK1, assay ID: Hs00938777_m1; CDKN1C, assay ID: Hs00175938_m1; E2F7, assay ID: Hs00987777_m1; FOXM1, assay ID: Hs01073586_m1) and Taqman ® Universal PCR master mix using the 7500 Fast Real-Time PCR system (all obtained from Applied Biosystems/ThermoFisher Scientific) according to the manufacturer’s recommendations.

Techniques: Expressing