肾癌中PPARα通过IKK改变NF-κB信号通路活性的实验研究  

PPARαmodulate NF-κB passway by IKK activation in renal cell carcinoma

作　　者：艾尼瓦尔·艾木都拉[1] 杨颖[1] 张瑞丽[1] 胡尔西旦·那斯尔[2] Ainiwaer·Aimudula;Yang Ying;Zhang Ruili;Huerxidan·Nasier(Cancer Centre,the First Affiliated Hospital of Xinjiang Medical University,Xinjiang Urumqi 830054,China;VIP Internal Medicine,the First Affiliated Hospital of Xinjiang Medical University,Xinjiang Urumqi 830054,China)

机构地区：[1]新疆医科大学第一附属医院肿瘤中心,新疆乌鲁木齐830054 [2]新疆医科大学第一附属医院VIP内科,新疆乌鲁木齐830054

出　　处：《现代肿瘤医学》2020年第10期1615-1619,共5页Journal of Modern Oncology

基　　金：新疆少数民族科技人才特殊培养项目(编号:2018D03013)。

摘　　要：目的:观察对舒尼替尼耐药的肾癌细胞系(A498、Caki-1、786-O)中PPARα调控NF-κB信号通路活性的作用机制。方法:首先培养对舒尼替尼耐药的三种肾癌细胞系(A498、Caki-1、786-O),上调和下调PPARα的表达后,用Western blot法和荧光素酶法,分别检测耐药和敏感细胞系中NF-κB p65的表达水平以及NF-κB活性。用RT-PCR和Westen blot法分别检测空白对照组和NF-κB+SN50(信号通路抑制组)中,PPARα的mRNA和蛋白表达水平。在稳定过表达PPARα和低表达PPARα时用Westen blot法分别检测耐药细胞株(A498)中IKK,P-IKK,P-NF-κB p65蛋白的表达水平。结果:舒尼替尼耐药的肾癌细胞系中,下调PPARα,抑制NF-κB p65的表达。抑制NF-κB信号通路,PPARα表达变化未出现显著性差异。上调PPARα基因表达,IKK、P-IKK、P-NF-κB p65蛋白的表达显著性升高。结论:PPARα可能通过改变上游基因IKK的表达来调控NF-κB信号通路活性。Objective:To study the mechanism of PPARαregulation role via NF-κB signal pathway in RCC.Methods:Western blot analysed NF-κB p65 expression levels and NF-κB activity assay in sunitinib-sensitive or resistant RCC cells(A498,Caki-1,786-O)with PPARαoverexpression or shRNA-mediated knockdown of PPARα.RT-PCR and Western blot analysed PPARαmRNA and relative protein expression levels in sunitinib-sensitive or resistant RCC cells.Western blot analysed IKK,P-IKK and P-NF-κB p65 protein expression levels with PPARαover expression or shRNA-mediated knockdown of PPARα.Results:NF-κB p65 was inhibited in sunitinib resistant renal cancer cell lines(786-O,caci-1 and a-498)with shRNA-mediated knockdown of PPARα.Inhibition of NF-κB signal pathway did not cause significant difference in PPARαexpression.The expressions of IKK,P-IKK and P-NF-κB p65 were significantly increased after the up-regulation of PPARα.Conclusion:PPARαmediates NF-κB activity via IKK expression alteration in RCC.

关 键 词：肾细胞癌 舒尼替尼耐药 PPARΑ IKK NF-ΚB信号通路 

分 类 号：R737.11[医药卫生—肿瘤]