泛素样含PHD和环指域1对胃癌细胞株SGC-7901生物学行为的影响  

Effect of ubiquitin like with PHD and ring finger domains 1 on biological behaviors of SGC -7901 cells

作　　者：屈芫[1] 王天明 严枫[2] Qu Yuan;Wang Tianming;Yan Feng(Department of Scientific Research, Nanfing Medical University Affiliated Cancer Hospital, Nanfing 210009, China;Jiangsu Key Laboratory of Molecular and Translational Cancer Research, Nanjing Medical University Affiliated Cancer Hospital, Nanjing 210009, China;School of Basic Medicine Sciences, Nanjing Medical University, Nanjing 211166, Chin)

机构地区：[1]南京医科大学附属肿瘤医院科研科,210009 [2]南京医科大学江苏省恶性肿瘤分子生物学与转化医学重点实验室,210009 [3]南京医科大学基础医学院,211166

出　　处：《中华实验外科杂志》2018年第4期634-637,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金（21475063）;江苏省医学创新团队（CXTDA2017017）

摘　　要：目的观察RNA干扰下调泛素样含PHD和环指域1（UHRF1）基因对人胃癌SGC-7901细胞生长、凋亡、侵袭转移和周期分布变化的影响。 方法合成针对UHRF1基因的小干扰RNA（siRNA）片段，转染至胃癌细胞株SGC-7901；实验分为3组：转染组（转染特异性siRNA）、阴性对照组（转染非特异性siRNA）、空白对照组；细胞转染24 h后用实时定量聚合酶链反应（Real-time PCR）和Western blot检测转染前后UHRF1的mRNA和蛋白的表达水平变化；细胞计数试剂盒（CCK-8）检测细胞增殖能力；Matrigel和Transwell小室实验检测细胞迁移侵袭能力，流式细胞术检测细胞凋亡及周期分布。 结果转染后，胃癌细胞株SGC-7901 UHRF1 mRNA的相对表达量为0.22±0.02，低于阴性对照组（P＝0.003）和空白对照组（P＝0.001），UHRF1蛋白的相对表达量为0.17±0.01，低于阴性对照组（P＝0.001）和空白对照组（P＝0.000）；细胞增殖能力显著降低（P＝0.000）;转染组细胞和非特异性阴性对照组侵袭细胞数分别为（117.3±4.2）个和（433.3±5.5）个（P＝0.000），迁移细胞数分别为（123.0±3.6）个和（377.3±4.7）个（P＝0.000），细胞的侵袭迁移能力显著减弱；转染组凋亡率为（16.36±0.84）%较阴性对照组（9.67±0.36）%、空白对照组（8.63±0.10）%明显增加（P＝0.002）；细胞周期分布改变显著，G0/G1期的细胞明显增多（P＝0.010）。 结论UHRF1能促进胃癌细胞株SGC-7901增殖，增强其迁移侵袭能力，抑制细胞凋亡。ObjectiveTo investigate the effects of knocking down ubiquitin like with PHD and ring finger domains 1 （UHRF1） gene using RNA interference technology on the growth, apoptosis, invasion, metastasis and cycle distribution of human gastric cancer SGC-7901 cells. MethodsThe small interfering RNA （siRNA） interference fragment targeting UHRF1 gene obtained by chemical synthesis was transfected into gastric cancer SGC-7901 cell line. The experiment was divided into three groups： knock-down group （transfected with specific siRNA）, negative control group （transfected with non-specific control siRNA）, and blank control group. At 24 h after transfection, real-time quantitative polymerase chain reaction （Real-time PCR） and Western blotting were used to detect the changes in mRNA and protein expression level of UHRF1; cell proliferation was detected by cell counting kit-8 （CCK-8） assay; the migration and invasion ability of SGC-7901 cells was analyzed using Matrigel and Transwell chamber tests; flow cytometry was used to detect the cell apoptosis and cell cycle distribution of SGC-7901 cells. ResultsAfter transfection, the relative expression of UHRF1 mRNA （0.22±0.02） was lower than negative control（P=0.003） and blank control（P=0.001） cell lines; the relative expression of UHRF1 protein （0.17±0.01） was lower than negative control（P=0.001） and blank control（P=0.000） cell lines; cell proliferation ability was decreased significantly （P=0.000）; the invaded cell number of knock-down group was 117.3±4.2 compared with 433.3±5.5 for negative control group（P=0.000）; in contrast, the migrated cell number of knock-down group was 123.0±3.6 compared with 377.3±4.7 for negative control group （P=0.000）; cell invasion and migration ability was decreased significantly; the cell apoptosis rate of knock-down group （16.36±0.84）% compared with negative control group （9.67±0.36）% and blank control group （8.63±0.10）% was decreased significantly （P=0.002）;

关 键 词：泛素样含PHD和环指域1 胃癌 增殖 侵袭 细胞周期 

分 类 号：R735.2[医药卫生—肿瘤]