Project description:Retinoblastoma Y79 cell line was treated with specific siRNA to silence EpCAM gene expression in vitro and RNA was isolated for microarray experiment to study the changes in the whole gene expression profiling in the Y79 cell line. The study identified 465 up-regulated genes (>1.0 fold) and 205 down-regulated genes (<0.5 fold) in response to knockdown of Ep-CAM.
Project description:The main objective of the study is to identify the de-regulated miRNAs in association with HMGA2 (oncogene) in the Retinoblastoma tumorus. The present experiment was carried out using Y79 (Retinoblastoma cell line), as the invitro model of Retinoblastoma tumor. The HMGA2 transcripts were silenced using siRNA and the post-silenced RB cells (at the end of 48 hours) were subjected to miRNA profiling using agilent platform.The key miRNAs de-regulated in this experiment has been validated using qRT-PCR and further their role has been evaluated by addition of specific antagomirs in RB cell lines (Y79 and Weri Rb 1).
Project description:The main objective of the study is to identify the de-regulated miRNAs in association with HMGA2 (oncogene) in the Retinoblastoma tumorus. The present experiment was carried out using Y79 (Retinoblastoma cell line), as the invitro model of Retinoblastoma tumor. The HMGA2 transcripts were silenced using siRNA and the post-silenced RB cells (at the end of 48 hours) were subjected to miRNA profiling using agilent platform.The key miRNAs de-regulated in this experiment has been validated using qRT-PCR and further their role has been evaluated by addition of specific antagomirs in RB cell lines (Y79 and Weri Rb 1). Agilent one-color experiment,Organism: Homo sapiens ,Agilent Human miRNA 8x15k Arrays AMADID: 021827 [Agilent miRNA labeling reagent and Hybridization Kit Cat # 5190-0408] RB cells (Y79) treated with anti-HMGA2 siRNA or with control
Project description:We hypothesized that Tiam1 is involved in invassiveness of retinoblastoma. The fuctional role of Tiam1 in cell progression and metastasis was tested by siRNA mediated knockdown of Tiam1 in retinoblastoma Y79 cells. The genes de-regulated in response to Tiam1 knockdown was analysed by cDNA microarray in which most of the actin cytoskeleton regulation proteins and apoptotic proteins were de-regulated. our results prove that Tiam1 modulates actin cytoskeleton and cell invasion in retinoblastoma. Retinoblastoma Y79 cells were treated with Tiam1 siRNA for 48hrs and cDNA microarray was performed to analyze the genes regulated by Tiam1 silencing compared to untreated Y79 cells. Experiments were performed in triplicates.
Project description:The differential gene expression study was conducted in the Anti - HMGA2 siRNA treated Retinoblastoma cell (Y79). The transient transfection in Y79 cells were performed using Anti-HMGA2 siRNA . The concentration and the time of incubation for the transfection was optimised to increase the silencing efficiency and reduce the off -target effects. After 48 hours of transfection the cells were collected and the whole genome cDNA microarray was performed to interpret the differential gene expression regulated by HMGA2 in Y79 cells. The results of the microarray data has been validated.
Project description:siRNAs have played a major role in cancer drug discovery, but their potential is hampered due to off-target effects. Thus, delivery systems like RNA aptamers have been used to enhance the specific delivery of these siRNAs to cancer stem cells. We report the efficacy of three different EpCAM aptamer siRNA chimeras, which were investigated both in vitro and in vivo for their ability to reduce cancer cell progression. Using these chimeras, we demonstrated specific gene knockdown in EpCAM positive cells which ultimately led to the apoptosis. To study the efficacy of these aptamer chimeras in vivo, retinoblastoma xenografts bearing NCC Rb C 51 cells were created for the first time. Systemic administration of these aptamer chimeras reduced tumour growth to about 50%. We further investigated the central Role of PLK1 in Cancer Progression and demonstrated the anti-cancer effects of targeted EpCAM siPLK1 approach. Using SILAC-Mass spectrometry analysis, we showed that silencing PLK 1 gene can lead to p53 mediated cell cycle arrest. Thus, we establish EpCAM-siRNA chimeras as potential markers for targeted anti-cancer applications, which paves a platform for efficient second line of therapies in addition to existing chemotherapy options.
Project description:We aimed to identify microRNAs which are altered upon EpCAM silencing.EpCAM is over expressed molecule in Retinoblastoma. Retinoblastoma Y 79 cell line was used as model to study the differential expression of microRNA in EpCAM siRNA treated cells.To validate technique we further confirmed deregulated expression of few microRNAs in Y 79 cells using Real Time PCR technique.
Project description:The differential gene expression study was conducted in the Anti - HMGA2 siRNA treated Retinoblastoma cell (Y79). The transient transfection in Y79 cells were performed using Anti-HMGA2 siRNA . The concentration and the time of incubation for the transfection was optimised to increase the silencing efficiency and reduce the off -target effects. After 48 hours of transfection the cells were collected and the whole genome cDNA microarray was performed to interpret the differential gene expression regulated by HMGA2 in Y79 cells. The results of the microarray data has been validated. One-color experiment,Organism: Human , Custom Whole Genome Human 8x60k designed by Genotypic Technology Pvt. Ltd. (Agilent-027114), Labeling kit: Agilent Quick-Amp labeling Kit (p/n5190-0442)
Project description:Gene expression analysis study in curcumin treated (20µM curcumin treated Y79 cells) and control Y79 cells (Suspension Y79 cells). Source were Y79 retinoblastoma cell line from human.
Project description:Retinoblastoma Y79 cell line was treated with scopoletin in vitro and RNA was isolated for microarray experiment to study the changes in the whole gene expression profiling in the Y79 cell line. The study identified 1675 up-regulated genes (>1.0 fold) and 1879 down-regulated genes (<-1 fold) in response to scopoletin treatment
