Notch1表达对肾癌细胞生物学行为的影响

发布时间：2018-08-31 12:00

【摘要】：目的探讨Notch1表达对肾癌细胞生物学行为的影响。方法通过Western印迹检测Notch1在人正常肾小管上皮细胞HK-2,人肾癌细胞786-O,Caki-1,769-P,GRC-1及ACHN中的表达;siRNA Notch1及siRNA NC转染肾癌细胞48 h后,Western印迹及RT-PCR检测转染情况,MTT法及Hoechst染色检测细胞活力及凋亡情况,Transwell法检测细胞迁移侵袭能力,Western印迹检测Bax,Bcl-2,基质金属蛋白酶(MMP)2,MMP9蛋白表达。结果 HK-2细胞中Notch1蛋白表达量显著低于5株肾癌细胞786-O,769-P,GRC-1,Caki-1及ACHN,且在Caki-1中Notch1表达量最高,因此作为后续实验的细胞株。siRNA Notch1及siRNA NC转染肾癌细胞48 h后,与siRNA NC组比较,siRNA Notch1组Notch1蛋白及mRNA表达量下调,细胞活力降低,细胞凋亡率提高,细胞侵袭数目减少,Bax表达量上调,Bcl-2、MMP2及MMP9表达量下调(P0.01)。结论 Notch1低表达能显著抑制肾癌细胞Caki-1的恶性生物学行为,可能与调控细胞凋亡蛋白及下调MMPs表达有关。
[Abstract]:Objective to investigate the effect of Notch1 expression on the biological behavior of renal carcinoma cells. Methods Western blot was used to detect the expression of Notch1 in normal renal tubular epithelial cells HK-2, 786-Ocari Caki-1769-PngGRC-1 and ACHN. After 48 h of transfection with siRNA NC, the expression of Notch1 in human renal cancer cells was detected by Western blot, RT-PCR assay and Hoechst staining. The ability of cell migration and invasion was detected by Transwell method and the expression of Bax,Bcl-2, matrix metalloproteinase 2 (MMP) 2) MMP9 protein was detected by Western blot. Results the expression of Notch1 protein in HK-2 cells was significantly lower than that in five renal cancer cell lines 786-O769-PfGRC-1Caki-1 and ACHN, and the highest expression of Notch1 was found in Caki-1 cells. Therefore, the cell line. SiRNA Notch1 and siRNA NC were transfected into renal cancer cells for 48 h. Compared with siRNA NC group, the expression of Notch1 protein and mRNA in siRNA Notch1 group was down-regulated, the cell viability was decreased, the apoptosis rate was increased, and the expression of Bcl-2mMP2 and MMP9 was down-regulated (P0.01). Conclusion the low expression of Notch1 can significantly inhibit the malignant biological behavior of renal cancer cell Caki-1, which may be related to the regulation of apoptosis protein and the down-regulation of MMPs expression.
【作者单位】：青海省第五人民医院肿瘤科;青海省第五人民医院骨科;
【分类号】：R737.11

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