Journal: Oncology Reports

Article Title: Sirtuin 7 promotes non-small cell lung cancer progression by facilitating G1/S phase and epithelial-mesenchymal transition and activating AKT and ERK1/2 signaling

doi: 10.3892/or.2020.7672

Figure Lengend Snippet: Overexpression/knockdown of SIRT7 in human NSCLC cells. (A) GFP analysis by fluorescence microscopy. (B) GFP analysis by flow cytometry. (C) SIRT7 overexpression or knockdown efficiency detected by RT-qPCR. (D) SIRT7 overexpression or knockdown efficiency detected by western blotting. The representative western blot images are presented in the upper panel. The relative level of SIRT7 protein in A549-SIRT7 (A549-mock served as a control) and H292-shSIRT7 (H292-shcontrol served as a control) NSCLC cells are presented in the lower panel. *P<0.05, Student's t -test, n=6 per group. SIRT7, sirtuin 7; NSCLC, non-small cell lung cancer; GFP, green fluorescent protein; reverse transcription-quantitative PCR.

Article Snippet: The pGEM/SIRT7 cloning plasmid carrying human SIRT7 coding sequence (CDS) (BC017305) was supplied by Sino Biological, Inc. Plasmids including pLenti6.3/IRES/GFP lentiviral plasmid-expressing blasticidin S deaminase and green fluorescent protein (GFP) as well as pLP1, pLP2 and VSVG lentiviral packing plasmids were supplied by Novobio Scientific, Inc. Lentiviruses including control shRNA lentivirus (LV-shcontrol) and SIRT7 shRNA (h) lentivirus (LV-shSIRT7) containing puromycin resistance gene were supplied by Santa Cruz Biotechnology, Inc.

Techniques: Over Expression, Fluorescence, Microscopy, Flow Cytometry, Quantitative RT-PCR, Western Blot, Real-time Polymerase Chain Reaction

Journal: Oncology Reports

Article Title: Sirtuin 7 promotes non-small cell lung cancer progression by facilitating G1/S phase and epithelial-mesenchymal transition and activating AKT and ERK1/2 signaling

doi: 10.3892/or.2020.7672

Figure Lengend Snippet: SIRT7 promotes growth and G1 to S-phase transition of human NSCLC cells. (A) CCK-8 analysis of NSCLC cell proliferation/growth. *P<0.05, two-way repeated measures ANOVA with LSD post hoc multiple comparisons, n=6 per group. (B and C) Colony formation assay. (B) The representative images. (C) The relative clonogenic ability of A549-SIRT7 (A549-mock served as a control) and H292-shSIRT7 (H292-shcontrol served as a control) NSCLC cells. *P<0.05, Student t -test, n=6 per group. (D-F) Tumor subcutaneous xenograft mouse model. (D) Tumor volume. *P<0.05, two-way repeated measures ANOVA with LSD post hoc multiple comparisons, n=6 per group. (E) The xenograft tumor images. (F) Tumor weight. *P<0.05, Student's t -test, n=6 per group. (G and H) Cell cycle analysis by flow cytometry. (G) The representative images. (H) The percentage of each cell cycle phase. *P<0.05, Student's t -test, n=6 per group. SIRT7, sirtuin 7; NSCLC, non-small cell lung cancer; CCK-8, Cell Counting Kit-8; ANOVA, analysis of variance; LSD, least significant difference.

Article Snippet: The pGEM/SIRT7 cloning plasmid carrying human SIRT7 coding sequence (CDS) (BC017305) was supplied by Sino Biological, Inc. Plasmids including pLenti6.3/IRES/GFP lentiviral plasmid-expressing blasticidin S deaminase and green fluorescent protein (GFP) as well as pLP1, pLP2 and VSVG lentiviral packing plasmids were supplied by Novobio Scientific, Inc. Lentiviruses including control shRNA lentivirus (LV-shcontrol) and SIRT7 shRNA (h) lentivirus (LV-shSIRT7) containing puromycin resistance gene were supplied by Santa Cruz Biotechnology, Inc.

Techniques: Sublimation, CCK-8 Assay, Colony Assay, Cell Cycle Assay, Flow Cytometry, Cell Counting

Journal: Oncology Reports

Article Title: Sirtuin 7 promotes non-small cell lung cancer progression by facilitating G1/S phase and epithelial-mesenchymal transition and activating AKT and ERK1/2 signaling

doi: 10.3892/or.2020.7672

Figure Lengend Snippet: SIRT7 facilitates human NSCLC cell in vitro migration and invasion as well as in vivo distant lung metastasis. (A and B) Wound healing assay. (A) The representative images. (B) The relative migratory ability of A549-SIRT7 (A549-mock served as a control) and H292-shSIRT7 (H292-shcontrol served as a control) NSCLC cells. (C and D) Transwell migration assay. (C) The representative images. (D) The relative migratory ability of the aforementioned cells. (E and F) Transwell invasion assay. (E) The representative images. (F) The relative invasive ability of the aforementioned cells. (G) The representative H&E staining images of lung tissues. (H) The number of tumor metastatic nodules in lung tissues. *P<0.05, Student's t -test, n=6 per group. SIRT7, sirtuin 7; NSCLC, non-small cell lung cancer; H&E, hematoxylin and eosin.

Article Snippet: The pGEM/SIRT7 cloning plasmid carrying human SIRT7 coding sequence (CDS) (BC017305) was supplied by Sino Biological, Inc. Plasmids including pLenti6.3/IRES/GFP lentiviral plasmid-expressing blasticidin S deaminase and green fluorescent protein (GFP) as well as pLP1, pLP2 and VSVG lentiviral packing plasmids were supplied by Novobio Scientific, Inc. Lentiviruses including control shRNA lentivirus (LV-shcontrol) and SIRT7 shRNA (h) lentivirus (LV-shSIRT7) containing puromycin resistance gene were supplied by Santa Cruz Biotechnology, Inc.

Techniques: In Vitro, Migration, In Vivo, Wound Healing Assay, Transwell Migration Assay, Transwell Invasion Assay, Staining

Journal: Oncology Reports

Article Title: Sirtuin 7 promotes non-small cell lung cancer progression by facilitating G1/S phase and epithelial-mesenchymal transition and activating AKT and ERK1/2 signaling

doi: 10.3892/or.2020.7672

Figure Lengend Snippet: SIRT7 activates AKT/ERK1/2 signaling and regulates the expression of G1-phase checkpoint molecules for G1 to S transition as well as EMT molecules for EMT induction in NSCLC cells. (A and B) Western blot analysis of AKT/ERK1/2, G1-phase checkpoint and EMT molecules. (A) The representative western blot images. (B) The relative level of p-AKT (T308)/AKT, p-AKT (S473)/AKT and p-ERK1/2/ERK1/2 as well as p21, p27, cyclin D1, cyclin E1, CDK2, CDK4, E-cadherin, N-cadherin, vimentin, Snail and Slug in A549-SIRT7 (A549-mock served as a control) or H292-shSIRT7 (H292-shcontrol served as a control) NSCLC cells. *P<0.05, Student's t -test, n=6 per group. (C) IHC analysis of AKT/ERK1/2, G1-phase checkpoint and EMT molecules in xenograft tumor tissues. The representative IHC images are presented. SIRT7, sirtuin 7; EMT, epithelial-mesenchymal transition; NSCLC, non-small cell lung cancer; AKT, protein kinase; ERK1/2, extracellular signal-regulated kinase 1/2; CDK, cyclin-dependent kinase; IHC, immunohistochemistry.

Article Snippet: The pGEM/SIRT7 cloning plasmid carrying human SIRT7 coding sequence (CDS) (BC017305) was supplied by Sino Biological, Inc. Plasmids including pLenti6.3/IRES/GFP lentiviral plasmid-expressing blasticidin S deaminase and green fluorescent protein (GFP) as well as pLP1, pLP2 and VSVG lentiviral packing plasmids were supplied by Novobio Scientific, Inc. Lentiviruses including control shRNA lentivirus (LV-shcontrol) and SIRT7 shRNA (h) lentivirus (LV-shSIRT7) containing puromycin resistance gene were supplied by Santa Cruz Biotechnology, Inc.

Techniques: Expressing, Western Blot, Immunohistochemistry