小分子RNA干预Notch1基因对肾癌透明细胞癌增殖的作用

发布时间：2018-06-20 10:39

  本文选题：Notch1基因 + RNA　； 参考：《泰山医学院》2014年硕士论文

【摘要】：目的 探究Notch1在肾癌组织中的表达及与肾癌临床病理之间的关联。进一步探究Notch1在肾透明细胞癌中的表达含量，利用小片段RNA(siRNA)干预方法沉默肾透明细胞癌中Notch1的表达，并观测其对在肾癌Caki-1细胞增殖的意义。探索Notch1基因在肾癌发生发展中的临床意义。 方法 搜集52例肾癌组织与30例非癌肾组织标本，利用荧光实时定量PCR（RT-PCR）和免疫组化方式剖析正常肾组织与肾癌组织中Notch1基因的表达差别，分析Notch1与临床病理参数的相关性联系。人体正常HK-2肾小管上皮细胞和肾细胞癌786O细胞和Caki-1体外培养使用。经过实时荧光定量PCR(RT-PCR)检测Notch1mRNA和蛋白印迹法（Western blot）检测蛋白的表达。设立特异性Notch1-siRNA干涉肾癌Caki-1细胞，用Lip2000转染试剂转染48时小，经过荧光定量PCR和印迹蛋白法分别检测Notch1mRNA和蛋白的表达含量不同，CCk-8与克隆试验了解其增殖情行的变化。 结果 Notch1于肾癌组织中的表现高于非肾癌肾组织（为80.8%比40.0%，P0.05)。Notch1的表达水平同样与肾癌的远处转移有关，发生转移的高于未转移的(P0.05)；直径≥7cm的癌中的Notch1含量要高于直径＜7cm的肿瘤(P0.05)。类似的，，Notch1于肾癌细胞株中的含量要高于正常肾小管细胞，具备统计学差异（P0.05)。干扰siRNA-notch1后可显著降低Caki-1细胞中Notch1mRNA和蛋白的表达，具有统计学差异（P0.05)，也检测出Caki-1细胞增殖本领的降低，差异也具备统计学差别（P0.05)。 结论 Notch1在肾癌中高表达大概与肾癌发生发展有相关性。同时，Notch1与肾癌的临床病理分期分级和肿瘤大小均有关性。肾透亮细胞癌组织中Notch1基因高表达，Notch1信号通路的下调可抑制肾细胞癌Caki-1细胞发生，调控肾细胞癌，对肾细胞癌的诊断和治疗有一定的指导意义。
[Abstract]:Objective to investigate the expression of Notch1 in renal cell carcinoma (RCC) and its relationship with clinicopathology. To further investigate the expression of Notch1 in renal clear cell carcinoma (RCC), the expression of Notch1 in renal clear cell carcinoma (RCC) was silenced by small fragment RNA-siRNAs intervention, and the significance of Notch1 expression in Caki-1 cell proliferation was observed. To explore the clinical significance of Notch1 gene in the development of renal cell carcinoma. Methods 52 cases of renal carcinoma and 30 cases of non-cancerous renal tissues were collected. The expression of Notch1 gene in normal renal tissues and renal cell carcinoma tissues was analyzed by real-time quantitative PCR and immunohistochemistry. To analyze the correlation between Notch1 and clinicopathological parameters. Normal HK-2 renal tubular epithelial cells and renal cell carcinoma 786O cells and Caki-1 were cultured in vitro. The expression of Notch1 mRNA and Western blot were detected by real-time fluorescence quantitative PCR RT-PCR. The specific Notch1-siRNA interference cell line Caki-1 was established and transfected with Lip2000 transfection reagent for 48 hours. The expression of Notch1 mRNA and protein was detected by fluorescence quantitative PCR and blotting method respectively. Results the expression of Notch1 in renal cell carcinoma was higher than that in non-renal cell carcinoma (80.8% vs 40.0%). The expression level of Notch1 was also related to the distant metastasis of renal carcinoma. The content of Notch1 in carcinoma with diameter 鈮

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