rb cell lines y 79  (ATCC)

Journal: Molecular oncology

Article Title: New retinoblastoma (RB) drug delivery approaches: anti-tumor effect of atrial natriuretic peptide (ANP)-conjugated hyaluronic-acid-coated gold nanoparticles for intraocular treatment of chemoresistant RB.

doi: 10.1002/1878-0261.13587

Figure Lengend Snippet: Fig. 3. Effects of treatment with ANP coupled HA-GNPs on tumor formation of etoposide resistant RB cells in in ovo chorioallantoic mem- brane (CAM) assays. Photographs of CAM tumors in situ and ruler measurements (in cm) of excised tumors revealing that tumors forming on the upper CAM 7 days after grafting of treated Y79-Etop (A) and WERI-Etop (B) cells were smaller compared to those arising from con- trol cells treated with PBS (ctr). (A) Histological analysis of paraffin sections of Y79-Etop CAM tumors by hematoxylin and eosin (HE) and Ki67 stains (brown signal). Black arrowheads exemplarily demarcate Ki67 positive cells positive cells. Scale bars: 600 lm. (C) Quantification of the total vessel area, vessel length, thickness and branching points of HA-GNP and ANP-HA-GNP treated CAM tumors compared to the controls as calculated by an IKOSA online software (KLM vision). The experiments were performed in triplicates. ANP, atrial natriuretic pep- tide; ANP-HA-GNP, ANP coupled HA-GNPs; GNP, gold nanoparticles; HA-GNP, hyaluronic acid coupled GNPs. Values represent means of independent animals SEM. *P < 0.05; **P < 0.01 statistical differences compared to the control group calculated by one-way ANOVA with Newman–Keuls post-test.

Article Snippet: The human retinoblastoma (RB) cell lines Y79 [28] (RRID: CVCL_1893) and WERI [29] (RRID: CVCL_1792), originally purchased from the Leibniz Institute DSMZ (German Collection of Microorganisms and Cell Cultures) were placed to our disposal by H. Stephan along with the RB cell line RB355 [30] (RRID: CVCL_S611), initially provided by K. Heise, and the corresponding etoposide-resistant RB cell lines Y79-Etop, WERI-Etop, and RB355-Etop.

Techniques: In Ovo, In Situ, Software, Control

Journal: Molecular oncology

Article Title: New retinoblastoma (RB) drug delivery approaches: anti-tumor effect of atrial natriuretic peptide (ANP)-conjugated hyaluronic-acid-coated gold nanoparticles for intraocular treatment of chemoresistant RB.

doi: 10.1002/1878-0261.13587

Figure Lengend Snippet: Fig. 4. Quantification of tumor formation from grafted retinoblastoma (RB) cells after gold nanoparticle treatment in chorioallantoic membrane (CAM) assays. Tumor formation capacity (A, B), CAM tumor weight (B, E) and CAM tumor size (C, F) of etoposide resistant RB cells was quantified after treatment with GNPs, HA-GNPs, ANP, or ANP-HA-GNPs. Upper row (A–C) shows the results for Y79-Etop cells and lower row (D–F) data for WERI-Etop cells. ANP, atrial natriuretic peptide; ANP-HA-GNP, ANP coupled HA-GNPs; ctr, PBS treated; GNP, gold nanoparticles; HA-GNP, hyaluronic acid coupled GNPs. Values are means of three independent experiments SEM. *P < 0.05; **P < 0.01 statistical differences compared to the control group calculated by one-way ANOVA with Newman–Keuls post-test.

Article Snippet: The human retinoblastoma (RB) cell lines Y79 [28] (RRID: CVCL_1893) and WERI [29] (RRID: CVCL_1792), originally purchased from the Leibniz Institute DSMZ (German Collection of Microorganisms and Cell Cultures) were placed to our disposal by H. Stephan along with the RB cell line RB355 [30] (RRID: CVCL_S611), initially provided by K. Heise, and the corresponding etoposide-resistant RB cell lines Y79-Etop, WERI-Etop, and RB355-Etop.

Techniques: Membrane, Control

Journal: International Journal of Molecular Sciences

Article Title: ADAM10 and ADAM17—Novel Players in Retinoblastoma Carcinogenesis

doi: 10.3390/ijms232012621

Figure Lengend Snippet: Verification of ADAM10 and ADAM17 knockdown efficiency. Efficient stable, lentiviral ADAM10 (shADAM10), ADAM17 (shADAM17) and ADAM10/17 (shADAM10/17) double knockdown was verified by quantitative real-time PCR ( a , d ) and western blot analyses in Y79 ( b , c ) and WERI-Rb1 cells ( e , f ). Indicated intensity ratios relative to ß-actin, used as a loading control, were calculated using MICRO MANAGER 1.4 software. Values are means of three independent experiments ± SEM. *** p < 0.001 and **** p < 0.0001 statistical differences compared to the control (ctr) group calculated by Student’s t -test.

Article Snippet: The human RB cell lines Y79 [ ] and WERI-Rb1 [ ], originally purchased from the Leibniz Institute DSMZ (German Collection of Microorganisms and Cell Cultures), were kindly provided by Dr. H. Stephan.

Techniques: Knockdown, Real-time Polymerase Chain Reaction, Western Blot, Control, Software

Journal: International Journal of Molecular Sciences

Article Title: ADAM10 and ADAM17—Novel Players in Retinoblastoma Carcinogenesis

doi: 10.3390/ijms232012621

Figure Lengend Snippet: Effects of ADAM10/17 single and double knockdown on cell viability and proliferation of RB cells. Stable ADAM10 (shADAM10), ADAM17 (shADAM17) and ADAM10/17 (shADAM10/17) knockdown decreases cell viability and proliferation levels of Y79 and WERI-Rb1 cells compared to control cells (ctr) as revealed by WST-1 assays ( a ), and BrdU stains ( b , c ). Values are means of three independent experiments ± SEM. ns p > 0.05; ** p < 0.01; *** p < 0.001 and **** p < 0.0001 statistical differences compared to the control (ctr) group calculated by Student’s t -test.

Article Snippet: The human RB cell lines Y79 [ ] and WERI-Rb1 [ ], originally purchased from the Leibniz Institute DSMZ (German Collection of Microorganisms and Cell Cultures), were kindly provided by Dr. H. Stephan.

Techniques: Knockdown, Control

Journal: International Journal of Molecular Sciences

Article Title: ADAM10 and ADAM17—Novel Players in Retinoblastoma Carcinogenesis

doi: 10.3390/ijms232012621

Figure Lengend Snippet: Effects of ADAM10/17 single and double knockdown on cell growth of RB cell lines. Stable ADAM10 (shADAM10), ADAM17 (shADAM17) and ADAM10/17 (shADAM10/17) knockdown decreases growth of Y79 ( a , c , e ) and WERI-Rb1 cells ( b , d , f ) compared to control cells (ctr) as revealed by growth curve analysis. Values are means of three independent experiments ± SEM. * p < 0.05; ** p < 0.01; *** p < 0.001 and **** p < 0.0001 statistical differences compared to the control (ctr) group calculated by Student’s t -test.

Article Snippet: The human RB cell lines Y79 [ ] and WERI-Rb1 [ ], originally purchased from the Leibniz Institute DSMZ (German Collection of Microorganisms and Cell Cultures), were kindly provided by Dr. H. Stephan.

Techniques: Knockdown, Control

Journal: International Journal of Molecular Sciences

Article Title: ADAM10 and ADAM17—Novel Players in Retinoblastoma Carcinogenesis

doi: 10.3390/ijms232012621

Figure Lengend Snippet: Effects of ADAM10/17 single and double knockdown on apoptosis and anchorage independent growth of RB cell lines. Stable ADAM10 (shADAM10), ADAM17 (shADAM17) and ADAM10/17 (shADAM10/17) knockdown increases caspase mediated apoptosis levels of Y79 and WERI-Rb1 cells compared to control cells (ctr) as revealed by DAPI cell counts ( a ) and caspase3/7 assays ( b ). Both RB cell lines show significantly reduced colony formation capacity ( c ) and colony sizes ( d , e ) after ADAM10 and ADAM 17 single or double knockdown as revealed by soft agarose assays. Values are means of three independent experiments ± SEM. ns > 0.05; * p < 0.05; ** p < 0.01; *** p < 0.001 and **** p < 0.0001 statistical differences compared to the control (ctr) group calculated by Student’s t -test.

Article Snippet: The human RB cell lines Y79 [ ] and WERI-Rb1 [ ], originally purchased from the Leibniz Institute DSMZ (German Collection of Microorganisms and Cell Cultures), were kindly provided by Dr. H. Stephan.

Techniques: Knockdown, Control

Journal: International Journal of Molecular Sciences

Article Title: ADAM10 and ADAM17—Novel Players in Retinoblastoma Carcinogenesis

doi: 10.3390/ijms232012621

Figure Lengend Snippet: Effect of lentiviral ADAM10/17 single and double knockdown on tumor formation of RB cell lines in vivo as revealed by chick chorioallantoic membrane (CAM) assays. Photographs of CAM tumors in situ (upper row) and ruler measurements (in cm) of excised CAM tumors (lower row) 7 days after inoculating the Y79 ( a ) and WERI-Rb1 ( b ) RB cell onto the CAM. Quantification of weight ( c ) and size ( d ) of CAM tumors developing from ADAM10 (shADAM10), ADAM17 (shADAM17) and ADAM10/17 (shADAM10/17) depleted Y79 and WERI-Rb1 cell lines in comparison the control cells (ctr). Values are means of at least three independent experiments ± SEM. ns p > 0.05; * p < 0.05; *** p < 0.001; and **** p < 0.0001 statistical differences compared to the control group calculated by Student’s t -test. ( e ) GFP antibody staining of stably GFP expressing Y79 and WERI-Rb1 cells in CAM tumor paraffin sections. Immunohistochemistry was revealed using diaminobenzidine (brown signal) and hematoxylin counterstaining (blue nuclei staining). Scale bars, 2 mm and 300 µM (magnified insets). #: blood vessel in CAM mesoderm, *: GFP-positive (GFP+) RB tumor cells, arrowheads: solid GFP-positive RB tumor mass.

Article Snippet: The human RB cell lines Y79 [ ] and WERI-Rb1 [ ], originally purchased from the Leibniz Institute DSMZ (German Collection of Microorganisms and Cell Cultures), were kindly provided by Dr. H. Stephan.

Techniques: Knockdown, In Vivo, Membrane, In Situ, Comparison, Control, Staining, Stable Transfection, Expressing, Immunohistochemistry

Journal: International Journal of Molecular Sciences

Article Title: ADAM10 and ADAM17—Novel Players in Retinoblastoma Carcinogenesis

doi: 10.3390/ijms232012621

Figure Lengend Snippet: Effects of stable, lentiviral ADAM10/17 single or double knockdown on RB cell migration in vivo. ( a ) Desmin stains of blood vessels (red) in representative CAM whole mounts showing extravasated GFP-labeled Y79 and WERI-Rb1 cells (green, marked with arrowheads) after ADAM17 knockdown (shADAM17) in comparison to control cells (ctr), scale bars: 100 μm. Real-time quantification of human GAPDH (hGAPDH) content, normalized against 18S RNA, in lower CAM punches 5 days after intravenous injection of ADAM10 (shADAM10), ADAM17 (shADAM17) and ADAM10/17 (shADAM10/17) depleted Y79 ( b ) and WERI-Rb1 ( c ) human RB cell lines and control cells (ctr). Values are means of at least three independent experiments ± SEM. ns p > 0.05; * p < 0.05 and *** p < 0.001 statistical differences compared to the control group calculated by Student’s t -test.

Article Snippet: The human RB cell lines Y79 [ ] and WERI-Rb1 [ ], originally purchased from the Leibniz Institute DSMZ (German Collection of Microorganisms and Cell Cultures), were kindly provided by Dr. H. Stephan.

Techniques: Knockdown, Migration, In Vivo, Labeling, Comparison, Control, Injection

Journal: International Journal of Molecular Sciences

Article Title: ADAM10 and ADAM17—Novel Players in Retinoblastoma Carcinogenesis

doi: 10.3390/ijms232012621

Figure Lengend Snippet: Analysis of L1CAM ectodomain shedding after ADAM10/17 single and double knockdown in Y79 and WERI-Rb1 cell lines. Western blot analysis of ADAM mediated L1CAM ectodomain (L1–200) shedding in cell culture supernatant of Y79 and WERI-Rb1 cells 72 h after ADAM10 (shADAM10), ADAM17 (shADAM17) single and ADAM10/17 (shADAM10/17) double knockdown in comparison to control cells (ctr). Representative western blots of L1 shedding upon ADAM knockdown ( a ) and quantification of the L1CAM ectodomain expression reveals a significant reduction of L1-200 shedding after ADAM10/17 single and double knockdown in the RB cell lines Y79 and WERI-Rb1 ( b ). Indicated intensity ratios relative to ß-actin, used as a loading control, were calculated using MICRO MANAGER 1.4 software. Values are means of three independent experiments ± SEM. * p < 0.05 and ** p < 0.01 statistical differences compared to the control (ctr) group calculated by Student’s t -test.

Article Snippet: The human RB cell lines Y79 [ ] and WERI-Rb1 [ ], originally purchased from the Leibniz Institute DSMZ (German Collection of Microorganisms and Cell Cultures), were kindly provided by Dr. H. Stephan.

Techniques: Knockdown, Western Blot, Cell Culture, Comparison, Control, Expressing, Software

Journal: International Journal of Molecular Sciences

Article Title: ADAM10 and ADAM17—Novel Players in Retinoblastoma Carcinogenesis

doi: 10.3390/ijms232012621

Figure Lengend Snippet: AKT and phospho-AKT (pAKT) expression levels after shRNA-mediated ADAM10/17 single (shADAM10 and shADAM17) or double knockdown (shADAM10/17) in the RB cell lines Y79 and WERI-Rb1 as revealed by western blot analysis. Representative western blots showing AKT and p-AKT expression levels ( a , e ) and quantification of the p-AKT/AKT ratio ( b – d , f – h ) after ADAM10/17 single and double knockdown in the RB cell lines Y79 and WERI-Rb1. Indicated intensity ratios relative to ß-actin, used as a loading control, were calculated using MICRO MANAGER 1.4 software. Values are means of three independent experiments ± SEM. ns p > 0.05 and * p < 0.05 statistical differences compared to the control group calculated by paired Student’s t -test.

Article Snippet: The human RB cell lines Y79 [ ] and WERI-Rb1 [ ], originally purchased from the Leibniz Institute DSMZ (German Collection of Microorganisms and Cell Cultures), were kindly provided by Dr. H. Stephan.

Techniques: Expressing, shRNA, Knockdown, Western Blot, Control, Software

Journal: International Journal of Molecular Sciences

Article Title: ADAM10 and ADAM17—Novel Players in Retinoblastoma Carcinogenesis

doi: 10.3390/ijms232012621

Figure Lengend Snippet: MiRNA-145, miRNA-152 and miRNA-365 expression levels in parental WERI-Rb1 and Y79 cell lines as revealed by real-time PCR ( a – c ). Values are means of at least three independent experiments ± SEM. * p < 0.05; ** p < 0.01; *** p < 0.001; and **** p < 0.0001 statistical differences compared to the control group calculated by Student’s t -test.

Article Snippet: The human RB cell lines Y79 [ ] and WERI-Rb1 [ ], originally purchased from the Leibniz Institute DSMZ (German Collection of Microorganisms and Cell Cultures), were kindly provided by Dr. H. Stephan.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Control