5-氮杂-2′-脱氧胞苷对宫颈癌细胞增殖及N-ras、c-myc基因表达的影响  

Effect of 5-Aza-2′-deoxycytidine on cell proliferation and expression of N-ras and c-myc gene of cervical cancer cells

作　　者：张谷香[1] 李书萍 李波 肖松舒[2] ZHANG Guxiang;LI Shuping;LI Bo;XIAO Songshu(Department of Obstetrics and Gynecology,the Fourth Hospital of Changsha,Changsha 410006,Hunan Province,China;Department of Obstetrics and Gynecology,the Third Xiangya Hospital of Central South University,Changsha 410000,Hunan Province,China)

机构地区：[1]长沙市第四医院妇产科,湖南长沙410006 [2]中南大学湘雅三医院妇产科,湖南长沙410000

出　　处：《新乡医学院学报》2021年第4期313-318,322,共7页Journal of Xinxiang Medical University

基　　金：湖南省科技创新计划项目(编号:2018JJ3782)。

摘　　要：目的探讨5-氮杂-2′-脱氧胞苷(5-Aza-CdR)对宫颈癌细胞增殖及N-ras、c-myc基因表达的影响。方法取对数生长期宫颈癌Hela细胞随机分为对照组、低剂量5-Aza-CdR组、中剂量5-Aza-CdR组及高剂量5-Aza-CdR组,分别给予终浓度为0、1、5、10μmol·L-1的5-Aza-CdR进行干预,采用四甲基偶氮唑盐法检测干预24、48、72、96 h时Hela细胞增殖能力,流式细胞仪检测干预24 h时Hela细胞凋亡率,免疫印迹和实时荧光定量聚合酶链反应法检测干预24 h时Hela细胞N-ras、c-myc蛋白及mRNA水平,酶联免疫吸附法检测干预24 h时Hela细胞上清液中转化生长因子-β1(TGF-β1)及前列腺素E 2(PGE 2)水平。结果干预24 h时,各组Hela细胞增殖能力比较差异无统计学意义(P>0.05);干预48、72、96 h时,低剂量5-Aza-CdR组、中剂量5-Aza-CdR组及高剂量5-Aza-CdR组Hela细胞增殖能力均显著低于对照组(P<0.05),高剂量5-Aza-CdR组Hela细胞增殖能力显著低于低剂量5-Aza-CdR组和中剂量5-Aza-CdR组(P<0.05);干预72、96 h时,中剂量5-Aza-CdR组Hela细胞增殖能力显著低于低剂量5-Aza-CdR组(P<0.05)。低剂量5-Aza-CdR组、中剂量5-Aza-CdR组及高剂量5-Aza-CdR组Hela细胞凋亡率显著高于对照组(P<0.05);低剂量5-Aza-CdR组Hela细胞凋亡率显著低于中剂量5-Aza-CdR组及高剂量5-Aza-CdR组(P<0.05);中剂量5-Aza-CdR组Hela细胞凋亡率显著低于高剂量5-Aza-CdR组(P<0.05)。低剂量5-Aza-CdR组、中剂量5-Aza-CdR组及高剂量5-Aza-CdR组Hela细胞中N-ras及c-myc蛋白和mRNA相对表达量显著低于对照组(P<0.05);低剂量5-Aza-CdR组Hela细胞中N-ras及c-myc蛋白和mRNA相对表达量显著高于中剂量5-Aza-CdR组及高剂量5-Aza-CdR组,中剂量5-Aza-CdR组Hela细胞中N-ras及c-myc蛋白和mRNA相对表达量显著高于高剂量5-Aza-CdR组(P<0.05)。低剂量5-Aza-CdR组、中剂量5-Aza-CdR组及高剂量5-Aza-CdR组Hela细胞上清液中TGF-β1水平显著高于对照组,PGE 2水平显著低于对照组(P<0.05);低剂量5-Objective To investigate the effect of 5-Aza-2′-deoxycytidine(5-Aza-CdR)on the expression of N-ras and c-myc gene and cell proliferation of cervical cancer cells.Methods The cervical cancer cells Hela in logarithmic growth phase were taken and randomly divided into the control group,low-dose 5-Aza-CdR group,medium-dose 5-Aza-CdR group,and high-dose 5-Aza-CdR group,the Hela cells in each group were treated with a final concentration of 0,1,5,10μmol·L-15-Aza-CdR.The tetramethylazolium salt method was used to determine the proliferation ability of Hela cells at 24,48,72,96 hours of intervention.Flow cytometry was used to detect the apoptosis rate of Hela cells at 24 hours of intervention,Western blot and real-time fluorescent quantitative polymerase chain reaction were used to detect the levels of N-ras,c-myc protein and mRNA in Hela cells at 24 hours of intervention.Enzyme linked immunosorbent assay was used to detect the levels of transforming growth factor-β1(TGF-β1)and prostaglandin E 2(PGE 2)in the supernatant of Hela cells at 24 hours of intervention.Results There was no significant difference in the proliferation ability of Hela cells among each group when intervened for 24 hours(P>0.05);the proliferation ability of Hela cells in the low-dose 5-Aza-CdR group,medium-dose 5-Aza-CdR group and high-dose 5-Aza-CdR group was significantly lower than that in the control group at 48,72,96 hours of intervention(P<0.05);the proliferation ability of Hela cells in the high-dose 5-Aza-CdR group was significantly lower than that in the low-dose 5-Aza-CdR group and midium-dose 5-Aza-CdR group(P<0.05);at 72,96 hours of intervention,the proliferation ability of Hela cells in the medium-dose 5-Aza-CdR group was significantly lower than that in the low-dose 5-Aza-CdR group(P<0.05).The apoptosis rate of Hela cells in the low-dose 5-Aza-CdR group,medium-dose 5-Aza-CdR group and high-dose 5-Aza-CdR group was significantly higher than that of the control group(P<0.05);the apoptosis rate of Hela cells in the low-dose 5-Aza-CdR

关 键 词：宫颈癌 5-氮杂-2′-脱氧胞苷 N-RAS c-myc 细胞增殖 

分 类 号：R737.33[医药卫生—肿瘤]