Journal: PLoS ONE

Article Title: Comparative mRNA and miRNA transcriptome analysis of a mouse model of IGFIR-driven lung cancer

doi: 10.1371/journal.pone.0206948

Figure Lengend Snippet: (A) Schematic of doxycycline inducible expression of IGFIR in the mouse lung via the SPC promoter. (B) Unsupervised hierarchical clustering dendrogram of RNA-Seq data from tumor (T, red) and non-transgenic normal lung (N, grey) samples. (C) Volcano plot of log2 fold changes and differential expression p values between tumor and normal lung tissue. (D) Pie chart illustrating percentage of genes up and down-regulated in IGFIR-driven tumors. (E) Dot plots of endogenous murine Igf1r (padj = 5.71E-11) and human IGFIR transgene (padj = 5.65E-249) mRNA expression following mapping to a hybrid genome. (F) Heatmap showing differential expression of markers of AT2 and Club cells as well as subtypes of non-small cell lung cancer. ADC, adenocarcinoma, SCC, squamous cell carcinoma, mADC, mucinous adenocarcinoma. Adjusted p-values for (C) and (E) were obtained from DESeq2.

Article Snippet: To further enrich for tumor cells, a biotinylated anti-human IGF1R antibody (1:10, clone REA271, Miltenyi Biotech) was added to the epithelial enriched fraction for 30 min on ice followed by positive selection and separation as above.

Techniques: Expressing, RNA Sequencing, Transgenic Assay, Quantitative Proteomics

Journal: Nature communications

Article Title: FoxK1 and FoxK2 in insulin regulation of cellular and mitochondrial metabolism.

doi: 10.1038/s41467-019-09418-0

Figure Lengend Snippet: Fig. 8 Schematic model of insulin regulation of FoxK1/K2 and FoxOs in cellular function. The FoxK Forkhead transcription factors translocate from the cytoplasm to nucleus reciprocally to the translocation of FoxO1. FoxK translocation to the nucleus is dependent on the Akt-mTOR pathway, while its localization to the cytoplasm in the basal state is dependent on GSK3. Once in the nucleus, FoxKs play important roles in regulation of genes, fatty acid oxidation, mitochondrial biogenesis, cell proliferation and survival. Where other unknown proteins (named here X) are in the FoxK and IR/IGF1R protein- protein complexes remains to be determined

Article Snippet: Mouse IR (MC224356) and IGF1R (MC224342) cDNA clones were from Origene.

Techniques: Cell Function Assay, Translocation Assay

Journal: OncoTargets and therapy

Article Title: microRNA-628 inhibits the proliferation of acute myeloid leukemia cells by directly targeting IGF-1R

doi: 10.2147/OTT.S192137

Figure Lengend Snippet: miR-628 directly targets IGF-1R in AML cells. Notes: ( A ) The putative wild-type (WT) and mutated (MUT) binding sites for miR-628 in the 3′-UTR of IGF-1R are shown. ( B ) miR-628 mimics or miR-NC and a luciferase plasmid carrying the WT or MUT miR-628-binding site were transfected into HL-60 and THP-1 cells. After 48 hours of transfection, the transfected cells were harvested and subjected to quantification of luciferase activity using a Dual-Luciferase Reporter Assay System. * P <0.05 vs miR-NC. ( C ) The expression levels of IGF-1R mRNA in the bone marrow samples were derived from 29 patients with AML and 23 healthy controls were detected with RT-qPCR. * P <0.05 vs miR-NC. ( D ) Spearman’s correlation analysis was used to examine the correlation between miR-628 and IGF-1R mRNA levels in patients with AML. r =-0.5393, P =0.0025. ( E , F ) RT-qPCR and Western blot analysis were performed to detect the expression levels of IGF-1R mRNA and protein, respectively, in miR-628-overexpressing HL-60 and THP-1 cells. Western blot analysis was repeated at least three times. * P <0.05 vs miR-NC. Abbreviations: AML, acute myeloid leukemia; IGF-1R, insulin-like growth factor 1 receptor; miR-NC, miRNA mimics negative control; RT-qPCR, reverse-transcription quantitative polymerase chain reaction; 3′-UTR, 3′-untranslated regions.

Article Snippet: The luciferase reporter plasmids harboring the wild-type (WT) and mutated (MUT) 3′-UTR of IGF-1R gene were designed and obtained from Shanghai GenePharma Co., Ltd.; these were referred as pmiR-IGF-1R-3′-UTR WT and pmiR-IGF-1R-3′-UTR MUT, respectively.

Techniques: Binding Assay, Luciferase, Plasmid Preparation, Transfection, Activity Assay, Reporter Assay, Expressing, Derivative Assay, Quantitative RT-PCR, Western Blot, Negative Control, Real-time Polymerase Chain Reaction

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: miR-483 is Down-Regulated in Polycystic Ovarian Syndrome and Inhibits KGN Cell Proliferation via Targeting Insulin-Like Growth Factor 1 (IGF1)

doi: 10.12659/MSM.897301

Figure Lengend Snippet: IGF1 is directly inhibited by miR-483. pre-miR-483, KGN cells transfected with pre-miR-483. miR-483 sponge, KGN cells transfected with miR-483 sponge. pre/sponge control, KGN cells transfected with the corresponding blank vector as a control. ( A ) The sequence AGGAGUGA in IGF1 3′UTR is predicted to be the binding site of miR-483. Five bases in the binding site are mutated to construct mut- IGF1 3′UTR. ( B ) Luciferase reporter assay shows that the activity of wildtype IGF1 3′UTR, but not mut- IGF1 3′UTR, can be inhibited by miR-483 overexpression compared to the control. ( C ) Luciferase reporter assay shows that the activity of wildtype IGF1 3′UTR, but not mut- IGF1 3′UTR, can be up-regulated by miR-483 sponge compared to sponge control. ( D ) IGF1 mRNA expression was inhibited by miR-483 overexpression and promoted by miR-483 sponge, as shown by qRT-PCR. ( E ) IGF1 protein level was inhibited by miR-483 overexpression and promoted by miR-483 sponge, as shown by Western blot. GAPDH is used as an internal reference. * P <0.05. ** P <0.01. IGF1, insulin-like growth factor1. CCNB1 – cyclin B1. CCND1 – cyclin D1. CDK2 – cyclin-dependent kinase 2.

Article Snippet: IGF1 -specific siRNA (RiboBio, Guangzhou, China) and the overexpression vector constructed using the coding sequence of IGF1 (GenBank No. NM_001111283) and pcDNA3.1 (Thermo Scientific, Carlsbad, CA) were used to alter IGF1 expression.

Techniques: Transfection, Control, Plasmid Preparation, Sequencing, Binding Assay, Construct, Luciferase, Reporter Assay, Activity Assay, Over Expression, Expressing, Quantitative RT-PCR, Western Blot

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: miR-483 is Down-Regulated in Polycystic Ovarian Syndrome and Inhibits KGN Cell Proliferation via Targeting Insulin-Like Growth Factor 1 (IGF1)

doi: 10.12659/MSM.897301

Figure Lengend Snippet: IGF1 promotes cell viability and proliferation in KGN cells. IGF1, KGN cells transfected with IGF1 overexpression vector. si-IGF1, KGN cells transfected with IGF1-specific siRNA. blank/si control, KGN cells transfection with the corresponding blank vectors as control groups. ( A ) IGF1 mRNA and protein expression was elevated in lesion ovary cortex of PCOS patients. ( B ) IGF1 mRNA and protein expression was altered by its overexpression vector or siRNA. ( C ) MTT assay shows KGN cell viability at 24, 48, and 72 h after transfection. ( D ) Pictures of colony formation. ( E ) Colony formation rate based on repeated colony formation assay experiment. ( F ) IGF1 expression in cells overexpressing miR-483 and IGF1 at the same time. ( G ) IGF1 overexpression in KGN cells transfected with miR-483 reverses miR-483-inhibited cell viability. * P <0.05. ** P <0.01. *** P <0.001. IGF1 – insulin-like growth factor 1. PCOS – polycystic ovarian syndrome.

Article Snippet: IGF1 -specific siRNA (RiboBio, Guangzhou, China) and the overexpression vector constructed using the coding sequence of IGF1 (GenBank No. NM_001111283) and pcDNA3.1 (Thermo Scientific, Carlsbad, CA) were used to alter IGF1 expression.

Techniques: Transfection, Over Expression, Plasmid Preparation, Control, Expressing, MTT Assay, Colony Assay

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: miR-483 is Down-Regulated in Polycystic Ovarian Syndrome and Inhibits KGN Cell Proliferation via Targeting Insulin-Like Growth Factor 1 (IGF1)

doi: 10.12659/MSM.897301

Figure Lengend Snippet: Insulin inhibits miR-483 and promotes IGF1 and KGN cell proliferation. ( A ) The miR-483 levels detected by qRT-PCR in cells treated with 0, 1, 10, and 100 ng/mL insulin for 48 h. ( B ) The IGF1 mRNA levels detected by qRT-PCR in cells treated with 0, 1, 10, and 100 ng/mL insulin for 48 h. ( C ) Insulin (100 ng/mL) promotes KGN cell viability compared to the untreated cells (control). Cell viability was detected with MTT method after 24, 48, or 72 h of insulin treatment. ( D ) Pictures of colony formation assay. ( E ) Colony formation rate was increased by insulin (100 ng/mL) detected by colony formation assay after 48 h of insulin treatment. * P <0.05. ** P <0.01. *** P <0.001. IGF1, insulin-like growth factor 1.

Article Snippet: IGF1 -specific siRNA (RiboBio, Guangzhou, China) and the overexpression vector constructed using the coding sequence of IGF1 (GenBank No. NM_001111283) and pcDNA3.1 (Thermo Scientific, Carlsbad, CA) were used to alter IGF1 expression.

Techniques: Quantitative RT-PCR, Control, Colony Assay

Journal: Breast Cancer Research : BCR

Article Title: Luminal-B breast cancer and novel therapeutic targets

doi: 10.1186/bcr2904

Figure Lengend Snippet: Targeted treatment in luminal-B breast cancer

Article Snippet: , Cixutumumab , ImClone , IGF-1R mAb , I/II , Cixutumumab and temsirolimus , Locally advanced/metastatic breast cancer progressed on one or two chemotherapy lines.

Techniques: Amplification, Expressing, Mutagenesis