dicer在肾透明细胞癌中的表达及其功能学分析word格式论文.docxVIP

The Expression and Functional Study of Dicer in Clear Cell Renal Cell CarcinomaAbstractObjective: Although multiple studies have shown that the deregulation of microRNA (miRNA) biogenesis machinery is involved in various human malignancies, this role has not been investigated in clear cell renal cell carcinoma (ccRCC). This study aims to determine the expression of Dicer, a key enzyme responsible for biogenesis of miRNA, and its biological roles in ccRCC, investigating its correlation with tumorigenesis and metastasis of ccRCC.Materials and Methods: The expression of Dicer in mRNA and protein levels were detected in human kidney tubule epithelial cell line (HKC), non-metastatic 786-O ccRCC cell line, and metastatic ACHN ccRCC cell line, as well as in 42 cases of ccRCC surgical specimens including 14 cases with distant metastasis and their corresponding adjacent normal renal tissues, by real-time quantitative polymerase chain reaction and western blot, respectively. Dicer expression levels in specimens were also measured by immunohistochemical staining. Liposome-mediated method was used to transfect siRNA sequence targeting Dicer and mock sequence as control into 786-O and ACHN cells. The knockdown efficiency of Dicer in cell lines after transfection was detected by western blot. The effects of Dicer on cell growth rate and cell cycles were detected by MTS assay and flow cytometric analyses, respectively, and the effects of Dicer on cell migration and cell invasion were detected by Boyden chamber Transwell assay.Results: Compared with HKC, Dicer expression levels were significantly downregulated in 786-O and ACHN ccRCC cell lines, with the metastatic ACHN ccRCC cell line even lower (all p0.05). Meanwhile, Dicer expression levels were significantly downregulated in ccRCC surgical specimens compared with adjacent normal renal tissues, with the metastatic ones further reduced (all p0.05), and Dicer mRNA levels were significantly correlated with overall TNM stage of ccRC