RNA干扰SALL4基因对儿童睾丸卵黄囊瘤细胞增殖和凋亡的影响  

The effect of RNA interference gene SALL4 on proliferation and apoptosis of testicular yolk sac tumor cells in children

作　　者：吕敏[1] 蔡祖仁 陈肖鸣[1] LYU Min;CAI Zuren;CHEN Xiaoming(Department of Pediatric Surgery,the First Affiliated Hospital of Wenzhou Medical University,Wenzhou 325015,China)

机构地区：[1]温州医科大学附属第一医院小儿外科

出　　处：《温州医科大学学报》2019年第12期885-888,893,共5页Journal of Wenzhou Medical University

基　　金：温州市科技计划项目（Y20140491）

摘　　要：目的:研究干扰SALL4基因对儿童睾丸卵黄囊瘤细胞的体外增殖、凋亡的影响。方法:通过构建SALL4干扰质粒载体pGPU6/GFP/Neo-SALL4 shRNA,转染儿童睾丸卵黄囊瘤细胞,并在mRNA和蛋白水平验证干扰效果,通过CCK-8、流式细胞术来研究其对儿童睾丸卵黄囊瘤细胞体外增殖、凋亡的影响。结果:成功构建载体并干扰SALL4表达后,荧光定量PCR结果示干扰组mRNA水平(24.0%±4.0%)较阴性对照组(93.0%±43.0%)显著下降(P<0.05),蛋白相对表达量(0.39±0.10)也较阴性对照组(1.02±0.09)显著降低(P<0.05),儿童睾丸卵黄囊瘤细胞的体外增殖活力显著降低,细胞凋亡率显著上升(P<0.05)。结论:RNA靶向干扰SALL4表达可抑制儿童睾丸卵黄囊瘤细胞增殖能力并诱导其凋亡,提示SALL4基因可能是儿童睾丸卵黄囊瘤一个潜在的治疗靶点。Objective:To study the effect of RNA interference gene SALL4 on the proliferation and apaptosis of testicular yolk sac tumor cells in children.Methods:The pGPU6/GFP/neo-sall4 shRNA was constructed as a vector to be transfected into pediatric testicular yolk sac tumor cells and interfere in SALL4.The interfering effect was verified by detecting mRNA and protein levels.CCK-8 and flow cytometry were employed to study the inhibition effect of interfering SALL4 in pediatric testicular yolk sac tumor cells.Results:After successfully transfecting the vector into pediatric testicular yolk sac tumor cells,the mRNA expression magnitudes of SALL4 interfered group was 24.0%±4.0%,as compared with Negative group(93.0%±43.0%),with lower levels of mRNA expression(P<0.05).And the protein expressions of SALL4 interfered group(0.39±0.10)had a lower protein expression than Negative group(1.02±0.09).Besides,the proliferation was significantly reduced,while the apoptosis rate was significantly increased in vitro(P<0.05).Conclusion:Targeting SALL4 RNA could inhibit the proliferation and induce apoptosis of pediatric testicular yolk sac tumor cells,suggesting that SALL4 may be a potential therapeutic target for pediatric testicular yolk sac tumor.

关 键 词：RNA干扰 SALL4 儿童 睾丸卵黄囊瘤 

分 类 号：R726[医药卫生—儿科]