吉林大学学报(医学版) ›› 2013, Vol. 39 ›› Issue (3): 534-538.doi: 10.7694/jldxyxb20130323

• 基础研究 • 上一篇    下一篇

hTERTp/TK/pGL3靶向抑制端粒酶活性及其对鼻咽癌干细胞的杀伤作用

杨柯柯,申聪香,文 忠,关小芳,赖肖芬,钱宇虹   

  1. 南方医科大学珠江医院耳鼻咽喉-头颈外科,广东 广州 510282
  • 收稿日期:2012-01-18 出版日期:2013-05-28 发布日期:2013-07-01
  • 通讯作者: 文 忠(Tel:020-62783136,E-mail:wenzhong60@163.com) E-mail:wenzhong60@163.com
  • 作者简介:杨柯柯(1986-),女,河南省禹州市人,医师,医学硕士,主要从事鼻咽癌基础与临床研究。
  • 基金资助:

    广东省科技计划项目资助课题(2008B030301344)

Targeted inhibition  of  hTERTp/TK/pGL3  on telomerase activity  and its killing effect on nasopharyngeal  cancer stem cells

YANG Ke-ke,SHEN Cong-xiang,WEN Zhong,GUAN Xiao-fang,LAI Xiao-fen,QIAN Yu-hong   

  1. Department of Otorhinolaryngology and Head-Neck Surgery,Zhujiang Hospital,Nanfang Medical University,Guangzhou 510282,China
  • Received:2012-01-18 Online:2013-05-28 Published:2013-07-01

摘要: [摘 要] 目的:探讨hTERTp/TK/pGL3载体靶向抑制端粒酶活性及其杀灭鼻咽癌CD133+干细胞的作用机制。方法:将已构建的hTERTp/TK/pGL3载体及其对照处理因素(CMV/TK/pGL3和TK/pGL3载体)转染至鼻咽癌CD133+干细胞、CD133-肿瘤细胞、人脐静脉内皮细胞(ECV)(对照组)和鼻咽癌SUNE未分选细胞中,采用Stretch PCR 法检测4种细胞端粒酶活性改变。MTT法测定CD133+干细胞和ECV细胞存活率。结果:CD133+鼻咽癌干细胞体外培养7 d后细胞逐渐增多,CD133+鼻咽癌干细胞体内成瘤实验阳性。CD133+鼻咽癌干细胞转染hTERTp/TK/pGL3或CMV/TK/pGL3后端粒酶活性降低;而ECV细胞端粒酶为阴性表达;CD133+鼻咽癌干细胞分别转染TK/pGL3、CMV/TK/pGL3和hTERTp/TK/pGL3后细胞存活率平均为(87.4±0.4)%、(20.5±0.4)%和(27.9±0.2)%,ECV对照细胞组转染TK/pGL3、CMV/TK/pGL3和hTERTp/TK/pGL3后细胞存活率平均为(90.7±0.1)%、(18.1±0.2)%和(86.2±0.1)%,hTERTp/TK/pGL3杀灭鼻咽癌CD133+干细胞效率明显高于ECV细胞组(P<0.01)。结论:hTERTp/TK/pGL3载体可以通过下调端粒酶活性靶向抑制端粒酶阳性的鼻咽癌CD133+干细胞。

关键词: 鼻咽肿瘤, CD133+肿瘤干细胞, 肿瘤靶向治疗, 端粒酶

Abstract: Abstract:Objective To explore the mechanism of  targeted inhibition of hTERTp/TK/pGL3 vector on  the telomerase activity and its killing effect on nasopharyngeal cancer stem cells.Methods The nasopharyngeal carcinaoma CD133+ stem cells,CD133-tumor cells, human umbilical vein endothelial cells (ECV cells) (control group) and nasopharyngeal carcinaoma SUNE unsorted cells were transfected by hTERTp/TK/pGL3 vector and its control vectors (CMV/TK/pGL3 and TK/pGL3).The telomerase avtivities of nasopharyngeal carcinaoma CD133+ stem cells and ECV cells were detected by Stretch PCR.The survival rates of CD133+stem cells and ECV cells were detected by  MTT method.Results After culture for 7 d in vitro,the number of nasopharyngeal carcinaoma CD133+ stem cells was increased, the tumorigenicity test of CD133+ stem cells  in vivo was positive.The telomerase activity of nasopharyngeal carcinaoma CD133+ stem cells was decreased after transfected by hTERTp/TK/pGL3 or CMV/TK/pGL3; the telomerase activity of EVC cells was negative.After transfected by TK/pGL3,CMV/TK/pGL3,and hTERTp/TK/pGL3,the  survival rates of CD133+ stem cells were 87.4%±0.4%,20.5%±0.4%,and 27.9%±0.2%,respectively. After transfected by TK/pGL3,CMV/TK/pGL3,and hTERTp/TK/pGL3,the  survival rates of ECV cells were 90.7%±0.1%,18.1%±0.2%,and 86.2%±0.1%,respectively.The efficiency of  hTERTp/TK/pGL3 vector  in  killing CD133+ stem cells was higher than that of  ECV cells(P<0.01).Conclusion hTERTp/TK/pGL3 vector can targetedly inhibit the telomerase-positive nasopharyngeal carcinaoma CD133+ stem cells by down-regulation of telomerase activity.

Key words: nasopharyngeal neoplasms, CD133+tumor stem cells, targeted tumor therapy, telomerase

中图分类号: 

  • R73-3