姜黄素联合索拉菲尼对肾癌细胞780-0中自噬及凋亡信号的影响  

Curcumin Exacerbates Sorafenib-induced Apoptosis in 780-0 Cells by Activating Autophagy Signaling

作　　者：赵建通[1] 刘文瞻[1] 万洪磊 路志民[1] 郭明涛[1] 王冰[1] 肖波[1] ZHAO Jiantong;LIU Wenzhan;WAN Honglei(Department of Urology,Handan First Hospital,Handan,056000)

机构地区：[1]邯郸市第一医院泌尿外科,邯郸056000

出　　处：《中国中西医结合肾病杂志》2023年第6期484-487,I0002,共5页Chinese Journal of Integrated Traditional and Western Nephrology

基　　金：河北省卫生健康委医学科学研究计划项目(No.20181642);河北省邯郸市科学技术研究与发展计划项目(No.1423108063-10)。

摘　　要：目的:探究天然化合物姜黄素与化疗药物索拉菲尼在体外的联合抗肾癌作用及其作用机制。方法:以肾癌细胞780-0为研究对象,采用MTT法测定细胞生长抑制率,采用荧光定量PCR(Real-time PCR)方法检测凋亡通路基因表达,采用4,6-二氨基-2-苯基吲哚(DAPI)染色法观察细胞凋亡小体数量,采用蛋白质免疫印迹法(Western blot)检测细胞凋亡通路及自噬通路相关蛋白表达。采用吖啶橙染色法检测细胞中自噬通量的水平。结果:与对照组相比,姜黄素及索菲拉尼呈浓度依赖性降低肾癌细胞780-0细胞活力。与索菲拉尼处理组相比,姜黄素联合索菲拉尼处理组中Bax、ATG5及LC3Ⅱ/LC3Ⅰ表达显著升高,Bcl-2的表达量显著降低,表明姜黄素能加剧索菲拉尼诱导的肾癌细胞780-0中凋亡及自噬信号的表达。吖啶橙染色结果进一步表明姜黄素加剧索菲拉尼诱导的肾癌细胞780-0自噬流水平。与此同时,在使用自噬的抑制剂3-甲基腺嘌呤抑制细胞自噬后,由姜黄素和索菲拉尼联合诱导的肾癌细胞780-0凋亡现象被抑制。结论:姜黄素可以加剧索拉菲尼诱导的肾癌细胞780-0凋亡,并且其可能是通过激活细胞内自噬信号来实现的。Objective:To investigate the combined anti-cancer effect of curcumin and sorafenib in vitro.Methods:Cell viability was determined by MTT assay;Real-time PCR was used to detect apoptosis signaling gene expression;DAPI staining was used to observe the number of apoptotic body;Western blot was used to detect the protein expression of apoptosis and autophagy pathway;Acridine orange staining was used to detect the autophagic flux.Results:Compared with the control group,the cell viability in the curcumin and sorafenib treatment groups was decreased in a dose dependent manner.The gene and protein expressions of Bax were increased in curcumin and sorafenib treatment groups,while the expression of Bcl-2 were decreased significantly,which indicates that curcumin and sorafenib treatment groups induced 780-0 cells apoptosis.Results of western blot and AO staining showed that curcumin aggravated sorafenib-induced autophagy in 780-0 cells.After inhibiting autophagy with 3-Methyladenine,the apoptosis of 780-0 cells induced by the combination of curcumin and sorafenib was inhibited.Conclusion:Curcumin aggravates sorafenib-induced apoptosis in 780-0 cells,and it was related to the activation of intracellular autophagy.

关 键 词：姜黄素 索拉菲尼 自噬 凋亡 肾癌细胞780-0 

分 类 号：R737.11[医药卫生—肿瘤]