Wnt信号通路和白细胞介素24对宫颈癌细胞放射敏感性的影响  

Effect of Wnt signaling pathway and interleukin-24 on radiosensitivity of cervical cancer cells

作　　者：王冬花[1] 姜祥兵[1] 龚世雄[1] 王中显 王平[1] WANG Dong-hua;JIANG Xiang-bing;GONG Shi-xiong;WANG Zhong-xian;WANG Ping(Department of Gynaecology and Obstetrics,Wuhan No.1 Hospital,Wuhan,430022,Hubei Province,China)

机构地区：[1]武汉市第一医院妇产科,湖北武汉430022

出　　处：《中国临床药理学杂志》2020年第2期158-161,共4页The Chinese Journal of Clinical Pharmacology

摘　　要：目的研究Wnt/β-连环蛋白(β-catenin)抑制剂(XAV939)在过表达白细胞介素-24(IL-24)的宫颈癌细胞放射敏感性中的作用。方法将细胞实验分为以下2种情况。(1)2组:对照组、IL-24组;(2)4组:IL-24组、XAV939+IL-24组、放射+IL-24组和XAV939+放射+IL-24组。对照组转染pAdtrack CMV;IL-24组转染pAdtrack CMV-IL-24;XAV939+IL-24组的Siha细胞转染pAdtrack CMV-IL-24后,用10μmol·L-1XAV939处理;放射+IL-24组的Siha细胞转染pAdtrack CMV-IL-24后,用8 Gy剂量(X射线)单次照射;XAV939+放射+IL-24组的Siha细胞转染pAdtrack CMV-IL-24后,用10μmol·L-1XAV93处理,再用8Gy剂量单次照射。以实时定量-PCR法检测细胞中白细胞介素-24(IL-24)基因表达水平,以蛋白质印迹法测定IL-24、β-catenin、c-Myc和裂解半胱氨酸天冬氨酸蛋白酶-3(Cleaved caspase-3)表达水平,以四甲基偶氮唑盐比色法检测细胞增殖水平,以流式细胞仪检测细胞凋亡。结果对照组和IL-24组的IL-24基因表达水平分别为1.00±0.00和2.65±0.21;这2组的IL-24蛋白表达水平分别为0.38±0.04和0.71±0.08;这2组的β-catenin蛋白表达水平分别为0.98±0.12和0.52±0.06;这2组的c-Myc蛋白表达水平分别为0.65±0.05和0.34±0.03,IL-24组与对照组比较,上述指标差异均有统计学意义(均P<0.05)。IL-24组、XAV939+IL-24组、Radiation+IL-24组和XAV939+Radiation+IL-24组的细胞存活率分别为(62.25±6.14)%,(41.30±3.26)%,(36.58±2.17)%和(27.18±3.69)%;这4组的细胞凋亡率分别为(16.62±1.47)%,(26.58±2.78)%,(29.47±2.43)%和(35.18±2.96)%;这4组的Cleaved caspase-3蛋白表达水平分别为0.13±0.02,0.21±0.03,0.32±0.04和0.48±0.05,XAV939+IL-24组、Radiation+IL-24组与IL-24组比较,或者XAV939+IL-24组、Radiation+IL-24组与XAV939+放射+IL-24组比较,上述指标的差异均有统计学意义(均P<0.05)。结论过表达IL-24可抑制宫颈癌细胞中Wnt信号通路激活,XAV939可以增加过表达IL-24的宫颈癌细胞放射敏感性,诱导细胞凋亡,抑制细胞增殖�Objective To investigate the role of Wnt/β-catenin inhibitor(XAV93)in the radiosensitivity of cervical cancer cells overexpressing interleukin-24(IL-24).Methods The cells were divided into two situation.(1)Two groups:control group,IL-24 group;(2)Four groups:IL-24 group,XAV939+IL-24 group,radiation+IL-24 group,XAV939+radiation+IL-24 group.Control group was transfected with p Adtrack CMV;IL-24 group was transfected with p Adtrack CMV-IL-24;XAV939+IL-24 group,Siha cells were transfected with pAdtrack CMV-IL-24 and with 10μmol·L-1 XAV939 treatment;radiation+IL-24 group,Siha cells were transfected with pAdtrack CMV-IL-24 then irradiated with a single dose of 8 Gy(X-ray);XAV939+radiation+IL-24 group,Siha cells were transfected with pAdtrack CMV-IL-24 and with 10μmol·L-1 was treated with XAV93 and then irradiated with a single dose of 8 Gy.Interleukin-24(IL-24)mRMA levels was detected by real-time quantitative-PCR.The expression levels of the IL-24,β-catenin,c-Myc,and cleaved cysteine-containing aspartate-specific proteases-3(Cleaved caspase-3)proteins were determined by Western blot.The cell proliferation was detected by MTT assay.The apoptosis rate was detected by flow cytometry.Results The expression levels of IL-24 mRNA in control group and IL-24 group were 1.00±0.00,2.65±0.21,respectively;the expression levels of IL-24 protein were 0.38±0.04,0.71±0.08,respectively;and expression levels ofβ-catenin protein were 0.98±0.12,0.52±0.06,respectively;the expression levels of c-Myc protein were 0.65±0.05,0.34±0.03,respectively.Comparison between IL-24 group and control group,the difference of the factors were significant(all P<0.05).The cell viability in IL-24 group,XAV939+IL-24 group,radiation+IL-24 group and XAV939+radiation+IL-24 group were(62.25±6.14)%,(41.30±3.26)%,(36.58±2.17)%,(27.18±3.69)%,respectively;the apoptosis rate in the four groups were(16.62±1.47)%,(26.58±2.78)%,(29.47±2.43)%,(35.18±2.96)%,respectively;expression levels of Cleaved caspase-3 protein were 0.13±0.02,0.21±0.03,0.32±0.0

关 键 词：宫颈癌 放射敏感性 白细胞介素24 WNT信号通路 

分 类 号：R97[医药卫生—药品]