番茄红素通过SIRT1/NF-κB轴抑制肾癌786-O细胞增殖且促进其凋亡  

Lycopene inhibits proliferation and promotes apoptosis of renal cancer 786-O cells through the SIRT1/NF-κB axis

作　　者：刘伟[1] 杨树芹 赵序雯 LIU Wei;YANG Shuqin;ZHAO Xuwen(Department of Pathology,Tengzhou Central People's Hospital,Tengzhou 277500,Shandong,China)

机构地区：[1]滕州市中心人民医院病理科,山东滕州277500

出　　处：《中国肿瘤生物治疗杂志》2023年第9期789-796,共8页Chinese Journal of Cancer Biotherapy

基　　金：济宁医学院教师科研项目(No.JYFC2018FKJ053)。

摘　　要：目的:探讨番茄红素通过沉默信息调节因子1(SIRT1)/核因子-κB(NF-κB)轴对肾癌786-O细胞增殖、凋亡的影响。方法:常规培养人正常肾细胞HK-2和人肾癌细胞786-O,实验分为对照组(0.1%DMSO)、顺铂组(40μg/mL)、番茄红素低质量浓度(2.5μg/mL)组、番茄红素高质量浓度(5μg/mL)组、番茄红素(5μg/mL)+EX527(SIRT1抑制剂)(3μmol/L)组。CCK-8法、克隆形成实验检测各组HK-2、786-O细胞的增殖能力,流式细胞术检测各组786-O细胞的凋亡,RH123、DCFH-DA染色分别检测各组786-O细胞的线粒体膜电位(MMP)、活性氧(ROS)水平,WB法检测各组786-O细胞中凋亡相关蛋白BAX、Bcl-2、C-casp3和SIRT1/NF-κB轴相关蛋白SIRT1、p-NF-κB蛋白的表达。786-O细胞移植瘤实验检测番茄红素低(5 mg/kg)、高质量浓度(20 mg/Kg)、顺铂(2 mg/kg)、番茄红素(20 mg/kg)+EX527(10 mg/kg)对移植瘤生长的影响,TUNEL法检测各组移植瘤组织中的细胞凋亡。结果:番茄红素呈剂量依赖性地抑制786-O细胞的增殖活性,番茄红素、顺铂均明显抑制786-O细胞的克隆形成能力且促进其凋亡,细胞中MMP损伤率升高而ROS水平降低,凋亡相关蛋白BAX、C-casp3表达均显著升高(均P<0.05)而Bcl-2表达下调(P<0.05),SIRT1表达显著升高(P<0.05)而p-NF-κB的表达显著降低(P<0.05),上述作用均可被EX527逆转;番茄红素、顺铂抑制786-O细胞移植瘤的生长且促进其细胞凋亡,其作用也能被EX527逆转。结论:番茄红素通过上调SIRT1、抑制NF-κB通路的激活进而抑制786-O细胞增殖且诱导其凋亡。Objective:To investigate the effects of lycopene on the proliferation and apoptosis of renal carcinoma 786-O cells through silent information regulator 1(SIRT1)/nuclear factor-kappa B(NF-κB)axis.Methods:Normal human kidney HK-2 cells and human renal cancer 786-O cells were conventionally cultured.The experiment included control group(0.1%DMSO),cisplatin group(40μg/mL),lycopene low concentration(2.5μg/mL)group,lycopene high concentration(5μg/mL)group,and lycopene high concentration(5μg/mL)+EX527(SIRT1 inhibitor)(3µmol/L)group.The proliferation capacity of 786-O cells in each treatment group was detected by CCK-8 method and clonal formation assay.The apoptosis of HK-2 and 786-O cells in each treatment group was detected by flow cytometry.The changes of mitochondrial membrane potential and reactive oxygen species(ROS)in 786-O cells in each treatment group were detected by RH123 and DCFH-DA staining,respectively.The expression of apoptosis-related proteins,BAX,Bcl-2,C-casp3 and SIRT1/NF-κB axis-related proteins SIRT1 and p-NF-Κb,in 786-O cells were detected by WB method.The effects of low concentration lycopene(5 mg/kg),high concentration lycopene(20 mg/Kg),cisplatin(2 mg/kg),and lycopene(20 mg/kg)+EX527(10 mg/kg)on the growth of 786-O cell transplanted tumor were observed.TUNEL method was used to detect the apoptosis in the tissues of transplanted tumors in each group.Results:Lycopene inhibited the proliferation activity of 786-O cells in a dose dependent manner.Lycopene and cisplatin significantly inhibited the clonogenesis ability of 786-O cells and promoted their apoptosis;moreover,after lycopene and cisplatin treatment,the MMP level was increased while ROS level was decreased,the expression of apoptosis-related proteins BAX and C-casp3 were significantly increased(all P<0.05),while the expression of Bcl-2 was down-regulated(P<0.05),the expression of SIRT1 was significantly increased(all P<0.05)and the expression of p-NF-κB was significantly decreased(all P<0.05).These effects could be reversed by EX527.

关 键 词：肾癌 786-O细胞 番茄红素 沉默信息调节因子1 核因子-κB 增殖 凋亡 

分 类 号：R737.11[医药卫生—肿瘤] R730.52[医药卫生—临床医学]