PART One:miR-134Functions As A Tumor Suppressor In Cell Proliferation And Epithelial-to-mesenchymal Transition By Targeting KRAS In Renal Cell Carcinoma Cells

Background:Aberrant microRNAs (miRNAs) are reported to contribute to the pathogenesis of most human malignancies. The miRNA miR-134has been found downregulated in renal cell carcinoma (RCC), but its function in the disease is unknown. The aims of this study were to detect the expression of miR-134in human RCC samples and explore its function in RCC cell lines.Methods:RT-PCR was used to quantify miR-134in human RCC samples. Assays for cell cycle, viability, migration, and invasion were performed to assess the phenotypic changes in RCC cells. A luciferase reporter assay was carried out to confirm whether KRAS is a direct target of miR-134. Western blot was used to identify the potential signaling pathways that had an impact on RCC cell growth and alterations of markers for EMT which affected metastasis by miR-134.Results:MiR-134was found downregulated in RCC samples (P＜0.05), while overexpression of miR-134suppressed proliferation (P<0.05) by triggering G1/G0cell cycle arrest (P＜0.05). Forced expression of miR-134could also inhibit migration (P<0.05) and invasion (P<0.05) by blocking EMT in RCC cell lines. KRAS was identified as a target of miR-134, and miR-134may act as a tumor suppressor via KRAS-related MAPK/ERK pathway other than PI3K/AKT signaling.Conclusion:miR-134may function as a tumor suppressor in cell proliferation and EMT by targeting KRAS in RCC cells. Background:Recent large-scale transcriptome analyses have found large numbers of transcripts, including that of long non-coding RNAs (IncRNAs), which are aberrant in various diseases, especially cancers. However, it is not clear whether IncRNAs are involved specifically in renal cell carcinoma (RCC). We investigated the expression patterns of IncRNAs in five RCC tumor samples (T) relative to those of matched adjacent non-tumor tissues (N) via microarray.Methods:A microarray with33,045IncRNA probes and30,215mRNA probes was used to identify deregulated IncRNAs in five RCC patients. Furthermore, we confirmed the relative expression levels of AK096725and ENST00000453068in70paired samples by quantitative reverse transcription polymerase chain reaction (RT-PCR).Results:The IncRNA microarray revealed27,279IncRNAs in RCC samples, of which480were significantly upregulated (P<0.05; T/N>1.5) and417were significantly downregulated (P<0.05; N/T>1.5) compared with the matched non-tumor samples. In addition,19,995mRNAs were detected, of which458were significantly upregulated (P＜0.05; T/N>1.5) and413were significantly downregulated (P<0.05; N/T>1.5). The expression level changes of AK096725(P=0.043) and ENST00000453068(P＜0.001) in70paired samples were in accord with the microarray data.Conclusions:The study uncovered expression patterns of IncRNAs in5RCC patients, as well as a number of aberrant IncRNAs and mRNAs in tumor samples compared with the non-tumor tissues. The revelation of an association between AK096725expression and RCC is especially noteworthy. These findings may help to find new biomarkers in RCC.