【摘要】 目的 探讨受体相互作用蛋白激酶1(RIPK1)对肾透明细胞癌(ccRCC)生物功能的影响及机制。方法 通过RNA干扰技术降低RIPK1在ccRCC中的表达后，光镜观察细胞形态变化，CCK-8法检测细胞增殖变化情况，Transwell细胞迁移实验检测细胞迁移能力的变化，流式细胞仪检测细胞发生凋亡、坏死水平，Western blot法检测cleaved caspase-3、混合谱系蛋白激酶样结构域(MLKL)和RIPK3表达变化。结果 小干扰RNA(siRNA)干扰48 h后，RIPK1-siRNA组细胞增殖吸光度值(0.563±0.042)低于NC-siRNA组(0.944±0.039;t=11.550,P=0.003);siRNA干扰72 h后，RIPK1-siRNA组吸光度值(0.408±0.019)低于NC-siRNA组(1.717±0.108;t=20.620,P<0.001)。与NC-siRNA组(72.000±12.120)比较，RIPK1-siRNA组(31.660±10.020)ccRCC细胞的迁移数量明显减少(t=4.442,P=0.011)。与NC-siRNA组(0.360±0.090、0.510±0.060、0.880±0.070)比较，RIPK1-siRNA组ccRCC细胞内cleaved caspase3、MLKL、RIPK3表达明显增加(0.780±0.070、1.490±0.100、1.680±0.130;t=6.380,P=0.003;t=8.656,P=0.001;t=14.150,P=0.001)。结论 RIPK1是ccRCC内调控细胞增殖、死亡及迁移能力的重要基因，抑制RIPK1能够促进ccRCC发生凋亡及坏死性凋亡。更多还原

【Abstract】 Objective To explore the mechanism of RNA interference on the expression of receptor-interacting protein kinase 1(RIPK1) gene in clear cell renal cell carcinoma. Methods After the expression of RIPK1 in ccRCC was reduced by RNA interference technology, the morphological changes of cells were observed by light microscopy, the changes of cell proliferation were detected by CCK-8 method, the changes of cell migration ability were detected by Transwell cell migration assay, the level of apoptosis and necrosis was detected by flow cytometry, and the expression changes of cleaved caspase-3, mixed-lineage kinase-like domain(MLKL) and RIPK3 was detected by Western blot. Results The cell proliferation absorbance values of in the RIPK1-small interfering RNA(siRNA) group(0.563±0.042) was lower than that in the NC-siRNA group(0.944±0.039; t=11.550,P=0.003). The absorbance values of in the RIPK1-siRNA group(0.408±0.019) was lower than that in the NC-siRNA group(1.717±0.108; t=20.620, P<0.001). The cell migration number of ccRCC in the RIPK1-siRNA group(31.660±10.020) was significantly lower than that in the NC-siRNA group(72.000±12.120; t=4.442, P=0.011). After siRNA interference with RIPK1, the expression levels of cleaved caspase3(0.780±0.070), MLKL(1.490±0.100), RIPK3(1.680±0.130) were significantly higher than those in the control group(0.360±0.090, 0.510±0.060, 0.880±0.070; t=6.380,P=0.003; t=8.656, P=0.001;t=14.150, P=0.001). Conclusion Our findings highlighted that RIPK1 was indispensable for clear cell renal cell carcinoma.更多还原