Project description:The aim of this analysis is searching the target gene of miR-135a-3p in ovarian cancer cell line. The gene expression of cell line with overexpressed miR-blank or miR-135a-3p was compared.
Project description:The aim of this analysis is searching the target gene of miR-135a-3p in ovarian cancer cell line. The gene expression of cell line with overexpressed miR-blank or miR-135a-3p was compared. Three ovarian cancer cell lines were enrolled this study. Those cells were transfected with miR-blank as control or miR-135a-3p.
Project description:We constructed a genome wide target profile of hsa-miR-503, hsa-miR-103, and hsa-miR-494 by sequencing RNA isolated from Ago2 immunoprecipitations and total RNA samples following transfection of the respective miRNA in mature duplex form Examination of mRNA levels in HeLa cells and Ago2 immunoprecipitations from HeLa cells following miR-503, miR-103, or miR-494 mature duplex or control siRNA transfection
Project description:MicroRNAs (miRNAs) are a class of endogenous small non-coding RNAs. They act as negative regulators of protein-coding gene expression at the post-transcriptional level via sequence-specific interaction with the 3' UTR of targeted mRNAs. A miR-135a dysregulation has been observed in various cancers, where a dual role of oncomiR or of tumor suppressor has been reported for the cancers profiled to date; however, knowledge is limited to explain these contentious data. In the present study, we investigate the regulation and mechanism of action of miR-135a, including identification of functionally targets that contributes to its actions, in prostate cancer progression. We demonstrate that the expression of miR-135a is regulated by androgens, in a time- and dose-dependent manner and identify miR-135a as a direct target gene of the androgen receptor AR. A functional androgen response element (ARE) is identified in the promoter region of the miR-135a gene and ChIP assay reveal AR protein binding to. Anti-androgen treatment and AR-targeted siRNA knockdown strategy suggest a novel function for miR-135a in AR signaling. In silico prediction, transcriptomic and proteomic combined analyses lead to identify ROCK-1/-2 as two miR-135a-targeted genes. Luciferase reporter assays indicate that these two proteins are miR-135a's direct targets. We finally demonstrate that miR-135a acts through ROCK-1/-2 pathways to affect migration and invasion cellular processes. MiR-135a expression level was lower in surgical cancerous samples compared to paired adjacent non-tumoral tissues and was gradually diminished consistent with cancer progression, suggesting the significance of miR-135a-mediated prostate cancer transformation. Our findings define miR-135a as a candidate tumor suppressor and potential prognostic marker in human prostate cancer.
Project description:MicroRNAs (miRNAs) are a class of endogenous small non-coding RNAs. They act as negative regulators of protein-coding gene expression at the post-transcriptional level via sequence-specific interaction with the 3' UTR of targeted mRNAs. A miR-135a dysregulation has been observed in various cancers, where a dual role of oncomiR or of tumor suppressor has been reported for the cancers profiled to date; however, knowledge is limited to explain these contentious data. In the present study, we investigate the regulation and mechanism of action of miR-135a, including identification of functionally targets that contributes to its actions, in prostate cancer progression. We demonstrate that the expression of miR-135a is regulated by androgens, in a time- and dose-dependent manner and identify miR-135a as a direct target gene of the androgen receptor AR. A functional androgen response element (ARE) is identified in the promoter region of the miR-135a gene and ChIP assay reveal AR protein binding to. Anti-androgen treatment and AR-targeted siRNA knockdown strategy suggest a novel function for miR-135a in AR signaling. In silico prediction, transcriptomic and proteomic combined analyses lead to identify ROCK-1/-2 as two miR-135a-targeted genes. Luciferase reporter assays indicate that these two proteins are miR-135a's direct targets. We finally demonstrate that miR-135a acts through ROCK-1/-2 pathways to affect migration and invasion cellular processes. MiR-135a expression level was lower in surgical cancerous samples compared to paired adjacent non-tumoral tissues and was gradually diminished consistent with cancer progression, suggesting the significance of miR-135a-mediated prostate cancer transformation. Our findings define miR-135a as a candidate tumor suppressor and potential prognostic marker in human prostate cancer.
Project description:We screened for target genes of hsa-mir-199a-1 and hsa-mir-4423 by transcriptome profiling. The miRNAs were overexpressed in HeLa cells by transient transfection of precursor miRNA expression plasmids. High-density Clariom D arrays for human were used for the gene expression analysis. This GEO entry provides the microarray data preprocessed according to the gene level workflow.