Journal: Cancers

Article Title: Mitochondrial VDAC1 Silencing in Urethane-Induced Lung Cancer Inhibits Tumor Growth and Alters Cancer Oncogenic Properties

doi: 10.3390/cancers16172970

Figure Lengend Snippet: PLGA-PEI-si-m/hVDAC1-B treatment of urethane-induced lung cancer reduced tumor size and number. ( A ) Schematic illustration of the study design: A/J mice, 7–8 weeks old, were injected i.p. with urethane (1 mg/gr body weight), and tumor development and treatment are indicated. ( B ) Photo of lungs from mice sacrificed at week 19, after exposure to urethane, with the arrows pointing to tumors. Following tracking of tumor development with MRI at week 18, treatment with PLGA-PEI-si-m/hVDAC1 or PLGA-PEI-si-NT (200 nM blood concentration) was started at week 19, and performed three times a week until the mice were sacrificed at week 38. ( C , F ) Representative H&E staining of paraffin-embedded, formaldehyde-fixed sections of the lungs after treatment with PLGA-PEI-si-m/hVDAC1-B or PLGA-PEI-si-NT. ( D , E ) Quantitative analysis of tumor area presented as a % of the total section area (PLGA-PEI-si-NT, n = 7, PLGA-PEI-si-m/hVDAC1, n = 8) ( D ), and the number of tumors (PLGA-PEI-si-NT n = 4 and PLGA-PEI-si-m/hVDAC1-B, n = 4) ( E ). Results reflect the mean ± SEM, *** p ≤ 0.001. ( F ) Enlargement of a selected area from the lung shown in C.

Article Snippet: Bovine serum albumin (BSA), poly (D, L-lactic-co-glycolic acid) (PLGA), polyethylenimine (PEI), polyvinyl alcohol (PVA), trypan blue, Triton X-100, Tween-20, hematoxylin, eosin, 4′,6-diamidino-2-phenylindole (DAPI), dimethyl sulfoxide (DMSO), and urethane were obtained from Sigma-Aldrich (St. Louis, MO, USA).

Techniques: Injection, Concentration Assay, Staining

Journal: Cancers

Article Title: Mitochondrial VDAC1 Silencing in Urethane-Induced Lung Cancer Inhibits Tumor Growth and Alters Cancer Oncogenic Properties

doi: 10.3390/cancers16172970

Figure Lengend Snippet: PLGA-PEI-si-m/hVDAC1-B treatment of urethane-induced lung cancer decreased VDAC1 expression level and inhibited tumor growth. ( A ) Schematic illustration of the study design: A/J mice, 7–8 weeks old, were injected i.p. with urethane (1 mg/gr body weight). Treatment with PLGA-PEI-si-m/hVDAC1 or PLGA-PEI-si-NT (350 nM) was started at week 21 (group A) or at week 23 (group B) and continued three times a week until the mice were sacrificed. MRI imaging was performed at weeks 18, 30, 32, and 33. Mice were sacrificed at week 33 (group A) and week 32 (group B). ( B ) Representative MRI images at week 33 (group A) of PLGA-PEI-si-NT- and PLGA-PEI-si-m/hVDAC1-B-treated mice. The arrows point to tumors. ( C ) Quantification of tumor volume from MRI images of group A at week 30 (PLGA-PEI-si-NT, n = 6, PLGA-PEI-si-m/hVDAC1-B, n = 7) and week 33 (PLGA-PEI-si-NT, n = 5, PLGA-PEI-si-m/hVDAC1-B, n = 5) and for group B at week 32 (PLGA-PEI-si-NT, n = 3, PLGA-PEI-si-m/hVDAC1-B, n = 4). ( D ) IF staining of VDAC1 in lung sections from PLGA-PEI-si-m/hVDAC1-B and PLGA-PEI-si-NT-treated mice. The squares point to the area enlarged below. ( E , F ) IHC staining for KI-67 using specific antibodies of PLGA-PEI-si-NT- and PLGA-PEI-si-m/hVDAC1-B-treated mice ( E ). For the sections, the lungs with tumors are shown with enlarged area below. KI-67 staining intensity was analyzed quantitatively (n = 7) ( F ). Results present the mean ± SEM, * p ≤ 0.05, *** p ≤ 0.001.

Article Snippet: Bovine serum albumin (BSA), poly (D, L-lactic-co-glycolic acid) (PLGA), polyethylenimine (PEI), polyvinyl alcohol (PVA), trypan blue, Triton X-100, Tween-20, hematoxylin, eosin, 4′,6-diamidino-2-phenylindole (DAPI), dimethyl sulfoxide (DMSO), and urethane were obtained from Sigma-Aldrich (St. Louis, MO, USA).

Techniques: Expressing, Injection, Imaging, Staining, Immunohistochemistry

Journal: Cancers

Article Title: Mitochondrial VDAC1 Silencing in Urethane-Induced Lung Cancer Inhibits Tumor Growth and Alters Cancer Oncogenic Properties

doi: 10.3390/cancers16172970

Figure Lengend Snippet: PLGA-PEI-si-m/hVDAC1-B treatment reduced expression of energy- and metabolism-related proteins. Representative IHC staining of lung sections of PLGA-PEI-si-m/hVDAC1-B- and PLGA-PEI-si-NT-treated mice using specific antibodies against Glut 1 ( A , B ), HK-I ( C , D ) and VDAC1 ( E , F ), shown in two different magnifications. Staining intensity was quantified as described in the Methods section. Results present the mean ± SEM (n = 5), * p ≤ 0.05.

Article Snippet: Bovine serum albumin (BSA), poly (D, L-lactic-co-glycolic acid) (PLGA), polyethylenimine (PEI), polyvinyl alcohol (PVA), trypan blue, Triton X-100, Tween-20, hematoxylin, eosin, 4′,6-diamidino-2-phenylindole (DAPI), dimethyl sulfoxide (DMSO), and urethane were obtained from Sigma-Aldrich (St. Louis, MO, USA).

Techniques: Expressing, Immunohistochemistry, Staining

Journal: Cancers

Article Title: Mitochondrial VDAC1 Silencing in Urethane-Induced Lung Cancer Inhibits Tumor Growth and Alters Cancer Oncogenic Properties

doi: 10.3390/cancers16172970

Figure Lengend Snippet: Effect of PLGA-PEI-si-m/hVDAC1-B treatment on the expression of CSC-related markers. ( A , B ) Representative lung sections IHC stained with specific antibodies against SOX2 ( A ) and ALDHA1 ( B ), showing enlarged areas from tumor-free and tumor tissues. ( C , D ) Representative IHC staining of stem cell markers CD44 and CD1133 in PLGA-PEI-si-NT- and PLGA-PEI-si-m/hVDAC1-B-treated mice ( C ) and their quantified staining intensity ( D ). Results reflect the mean ± SEM (n = 3), ** p ≤ 0.01, *** p ≤ 0.001.

Article Snippet: Bovine serum albumin (BSA), poly (D, L-lactic-co-glycolic acid) (PLGA), polyethylenimine (PEI), polyvinyl alcohol (PVA), trypan blue, Triton X-100, Tween-20, hematoxylin, eosin, 4′,6-diamidino-2-phenylindole (DAPI), dimethyl sulfoxide (DMSO), and urethane were obtained from Sigma-Aldrich (St. Louis, MO, USA).

Techniques: Expressing, Staining, Immunohistochemistry

Journal: Cancers

Article Title: Mitochondrial VDAC1 Silencing in Urethane-Induced Lung Cancer Inhibits Tumor Growth and Alters Cancer Oncogenic Properties

doi: 10.3390/cancers16172970

Figure Lengend Snippet: Urethane treatment of A/J mice may induce both small and non-small lung cancer. ( A ) Representative H&E staining of formaldehyde-fixed paraffin-embedded sections from A/J mice lungs treated with PLGA-PEI-si-NT, showing two different morphologies as visualized in images at three different magnifications. ( B , C ) Representative lung sections of PLGA-PEI-si-NT-treated mice, IHC stained using specific antibodies against KI-67 ( B ) and synaptophysin ( C ), known to be highly expressed in SCLC. The squares show the areas from the high and low IHC-stained tumors that were magnified 25-fold in the images shown on the right. ( D ) Quantitative analysis of the indicated proteins’ staining intensities. Results reflect the mean ± SEM (n = 3), **** p ≤ 0.0001.

Article Snippet: Bovine serum albumin (BSA), poly (D, L-lactic-co-glycolic acid) (PLGA), polyethylenimine (PEI), polyvinyl alcohol (PVA), trypan blue, Triton X-100, Tween-20, hematoxylin, eosin, 4′,6-diamidino-2-phenylindole (DAPI), dimethyl sulfoxide (DMSO), and urethane were obtained from Sigma-Aldrich (St. Louis, MO, USA).

Techniques: Staining