双孔钾离子通道task-3在肾透明细胞癌中的表达及task-3抑制剂对786-0肾癌细胞增殖影响的分析-expression of double-hole potassium channel task - 3 in renal clear cell carcinoma and analysis of the effect of task - 3 inhibitor on 786 - 0 renal cell carcinoma cell p.docxVIP

福建医 福建医科大学医学硕士研究生学位论文 Abstract Objective: To investigate the expression of TASK-3 in clear cell renal cell carcinoma(ccRCC) and 786-0 cell line. Meanwhile, to analyse the effect to the 786-0 proliferation after using the inhibitor of TASK-3, and demonstrate the roles in ccRCC. Methods: TASK-3 mRNA and protein expressions were detected in 30 freeze-stored and paraffin-embedded renal carcinoma tissues and normal kidney tissues by real-time fluorescence quantitative reverse transcription PCR (RFQ-RT-PCR) and immunohistochemistry(IHC), respectively. Meanwhile, TASK-3 location on 786-0 cell line was tested by immunofuorescence and the cell growth was analysed by MTS assay after adding different concentration of TASK-3 inhibitor spermine. Results: The relative expression of TASK-3 mRNA in ccRCC tissues were significantly higher than that in normal kidney tissues (M ??Q were (2.33 ??8.89)×10-4, (0.13 ??3.90)×10-4, respectively(Shapiro-Wilcoxon test，p=0.029); Analogously, 83.33%(25/30) of ccRCC tissues were detected overexpression of TASK-3 protein, which were detected in only 13.33%(4/30) of normal kidney tissues, and the remaining 26 normal kidney tissues were detected low expression and even without expression at all(Fisher exact test, P ＜0.001). Both of the strong TASK-3 mRNA and protein expression were not distinctly correlated with clinicopathological features, such as TNM-staging and Fuhrman-grading (P＞0.05). Immunofuorescence experiment in cultured 786-0 renal carcinoma cell line showed that the cell line presented TASK-3 positive immunpositivity both on membrane and nucleus. Cell survival decreased with the increasing concentration of relative TASK-3 inhibitor spermine by MTS assay(Dunnett’S T3 test, P＜0.05). Conclusion: The results indicate that TASK-3 is an important mediator of RCC occurrence and progression, and offers potential as a molecular target for RCC therapies. Keywords: clear cell renal cell carcinoma, TASK-3, expression, PCR, I