牛磺酸上调基因1对人绒毛膜癌JEG-3细胞增殖及迁移和凋亡的影响  

Effects of long non-coding RNA TUG1 on proliferation,migration and apoptosis of JEG-3

作　　者：谢环[1] 田洁[1] 黄娅芬 XIE Huan;TIAN Jie;HUANG Yafen(Department of Obstetrics and Gynecology,the Second People's Hospital of Yichang, the Second People's Hospital of Three Gorges University,Yichang 443000,China;Department of Obstetrics and Gynecology,Huangshi Maternity and Child Health Care Hospital of Edong Healthcare Group,Huangshi 435000,China)

机构地区：[1]宜昌市第二人民医院三峡大学第二人民医院妇产科,湖北宜昌443000 [2]鄂东医疗集团黄石市妇幼保健院妇产科,湖北黄石435000

出　　处：《中华实用诊断与治疗杂志》2019年第1期14-17,共4页Journal of Chinese Practical Diagnosis and Therapy

基　　金：湖北省自然科学基金(2015CFB574)

摘　　要：目的探讨牛磺酸上调基因1(taurine-upregulated gene 1,TUG1)在人绒毛膜癌JEG-3细胞增殖、迁移和凋亡中的作用。方法采用小干扰RNA(small interfering RNA,siRNA)技术构建TUG1慢病毒载体。将对数生长期人绒毛膜癌JEG-3细胞随机分为空白对照组(不作任何处理)、阴性对照组(转染阴性对照靶序列慢病毒载体)和转染组(转染siRNA-TUG1慢病毒载体)。采用实时荧光定量PCR法检测3组TUG1mRNA、caspase-3mRNA、caspase-9mRNA相对表达量,采用Western blot法检测3组TUG1、caspase-3、caspase-9蛋白相对表达量,采用Transwell小室试验检测3组培养24h细胞侵袭能力,采用CCK-8法检测3组细胞培养72h吸光度值,采用流式细胞术检测3组培养72h细胞凋亡率。结果构建siRNA-TUG1载体的慢病毒滴度为3×108 TU/mL;转染72 h,转染组TUG1 mRNA、caspase-3mRNA、caspase-9mRNA相对表达量(0.42±0.02、0.31±0.01、0.21±0.01)低于阴性对照组(1.09±0.03、1.29±0.03、1.02±0.01)和空白对照组(1.01±0.01、1.02±0.02、1.00±0.01)(P<0.05),TUG1、caspase-3、caspase-9蛋白相对表达量(0.24±0.07、0.49±0.23、0.11±0.02)低于阴性对照组(7.32±0.05、10.12±1.47、9.98±2.78)和空白对照组(6.08±0.19、9.88±0.93、9.91±1.44)(P<0.05);培养24h,转染组迁移细胞数[(52.11±4.09)个]较阴性对照组[(142.11±14.02)个]和空白对照组[(131.32±18.39)个]少(P<0.05);培养72h,转染组细胞吸光度值(24.27±3.11)较阴性对照组(51.12±2.47)和空白对照组(60.34±1.24)低(P<0.05),细胞凋亡率[(35.21±13.31)%]较阴性对照组[(3.09±1.13)%]和空白对照组[(4.21±1.24)%]高(P<0.05);阴性对照组各观察指标与空白对照组比较差异均无统计学意义(P>0.05)。结论 TUG1沉默表达慢病毒载体可明显抑制人绒毛膜癌JEG-3细胞增殖和侵袭,促进细胞凋亡。Objective To investigate the role of taurine-upregulated gene 1(TUG1)in the proliferation,migration and apoptosis of human choriocarcinoma JEG-3cells.Methods TUG1 lentivirus vectors were constructed by small interfering RNA technology.JEG-3cells in logarithmic growth phase were randomly divided into blank control group(without any treatment),negative control group(transfected by lentivirus particle empty vector)and transfection group(transfected by siRNA-TUG1 lentivirus vector).The relative mRNA and protein expressions of TUG1,caspase-3and caspase-9were detected by real-time PCR and Western blot in three groups.Transwell assay was used to detect the invasive ability of cells after 24 hof culturing,CCK-8method was used to detect the absorbance value after 72 hof culturing,and flow cytometry was used to detect the cell apoptosis after 72 hof culturing.Results The lentivirus titer of the constructed siRNA-TUG1 vector was 3×108 TU/mL.After 72 hof transfection,the relative mRNA expressions of TUG1,caspase-3and caspase-9were significantly lower in transfection group(0.42±0.02,0.31±0.01,0.21±0.01)than those in negative control group(1.09±0.03,1.29±0.03,1.02±0.01)and blank control group(1.01±0.01,1.02±0.02,1.00±0.01)(P<0.05),while the relative protein expressions of TUG1,caspase-3and caspase-9(0.24±0.07,0.49±0.23,0.11±0.02)were significantly lower in transfection group than those in negative control group(7.32±0.05,10.12±1.47,9.98±2.78)and blank control group(6.08±0.19,9.88±0.93,9.91±1.44)(P<0.05).The count of migrated cells was significantly less in transfection group(52.11±4.09)than that in negative control group(142.11±14.02)and blank control group(131.32±18.39)after 24 hof culturing(P<0.05).The cell absorbance value was significantly lower in transfection group(24.27±3.11)than that in negative control group(51.12±2.47)and blank control group(60.34±1.24)after 72 hof culturing(P<0.05).The apoptotic rate was significantly higher in transfection group((35.21±13.31)%)than that in negative control

关 键 词：绒毛膜癌 长链非编码RNA 牛磺酸上调基因1 增殖 凋亡 迁移 

分 类 号：R737.33[医药卫生—肿瘤]