SATB2基因增强槲皮素抑制肾癌细胞生长的机制

Mechanism of SATB2 gene enhances quercetin on growth inhibition of renal cell carcinoma

目的:探讨核基质蛋白特异AT序列结合蛋白2(special AT-rich sequence-binding protein 2,SATB2)基因及槲皮素对肾癌细胞活力、侵袭能力和凋亡的影响及机制。方法:人肾癌786-O细胞分为空白组、重组体pcDNA3.1-SATB2组、槲皮素组和pcDNA3.1-SATB2+槲皮素组,其中pcDNA3.1-SATB2转染参照Lipofectamine 2000说明。各组细胞处理48 h后,通过CCK-8,Transwell小室及流式细胞术分别检测细胞活力、侵袭能力和凋亡。Western印迹法检测SATB2,STAT3,p-STAT3,MMP-2和Bcl-2蛋白表达。结果:转染pcDNA3.1-SATB2的786-O细胞SATB2蛋白表达明显升高(P<0.05)。pcDNA3.1-SATB2及不同浓度槲皮素均可抑制786-O细胞活力,且槲皮素可呈现浓度和时间依赖性抑制细胞活力(P<0.05)。pcDNA3.1-SATB2及槲皮素均可抑制786-O细胞侵袭能力,诱导凋亡,下调p-STAT3,MMP-2和Bcl-2表达,而联合使用pcDNA3.1-SATB2和槲皮素对786-O细胞侵袭能力、凋亡及p-STAT3,MMP-2和Bcl-2表达影响更明显(P<0.05)。结论:过表达SATB2可增强槲皮素对肾癌细胞活力和侵袭能力抑制及凋亡促进作用,机制与抑制STAT3信号通路有关。

Objective: To investigate the effect of special AT-rich sequence-binding protein 2 (SATB2) gene and quercetin on the activity, invasion and apoptosis of renal cell carcinoma (RCC) and its mechanism. Methods: Human renal carcinoma 786-O cells were divided into blank group, recombinant pcDNA3.1-SATB2 group, quercetin group and pcDNA3.1-SATB2+quercetin group, in which pcDNA3.1-SATB2 was transfected according to Lipofectamine 2000. After 48 hours in culture, CCK-8, Transwell chamber and flow cytometry were used to detect cell viability, invasion and apoptosis. Western blotting was used to detect the expression of SATB2, STAT3, p-STAT3, MMP-2 and Bcl-2 protein. Results: The expression of SATB2 protein in 786-O cells transfected with pcDNA3.1-SATB2 increased significantly (P<0.05). pcDNA3.1-SATB2 and quercetin with different concentrations inhibited 786-O cell viability, quercetin also inhibited cell viability in a concentration and time dependent manner (P<0.05).pcDNA3.1-SATB2 and quercetin could inhibit the invasive ability of 786-O cells, induce apoptosis and downregulate the expression of p-STAT3, MMP-2 and Bcl-2. The combination of pcDNA3.1-SATB2 and quercetin had more significant effects on the invasive ability, apoptosis and the expression of p-STAT3, MMP-2 and Bcl- 2 of 786-O cells (P<0.05). Conclusion: Overexpression of SATB2 can enhance inhibition of cell viability and invasion and apoptosis promoting of quercetin on 786-O cells, which may be related to the inhibition of STAT3 signaling pathway.