人端粒酶逆转录酶基因转染U2OS细胞对其基质金属蛋白酶2活性的影响

J4 ›› 2009, Vol. 35 ›› Issue (5): 812-816.

人端粒酶逆转录酶基因转染U2OS细胞对其基质金属蛋白酶2活性的影响

姜成威¹, 杨媚², 石英爱³, 于晓霞³, 吴珊³

1. 吉林大学中日联谊医院病理科|吉林长春 130033；2. 吉林大学第二医院病理科|吉林长春 130041；3.吉林大学基础医学院病理生物学教育部重点实验室|吉林长春 130021

收稿日期:2009-01-08 出版日期:2009-09-28 发布日期:2009-09-28
通讯作者: 吴珊 E-mail:wushan@jlu.edu.cn
作者简介:姜成威（1981-）|男|吉林省辽源市人|医师|医学硕士|主要从事肿瘤病理学的研究。
基金资助:
教育部高校优秀青年教师奖励计划资助课题（201182）；吉林省科技厅白求恩专项基金资助课题(200705369)

Effect of |U2OS cells transfected with hTERT on activity of matrix metalloproteinase-2

JIANG Cheng-Wei¹, YANG Mei², DAN Yang-Ai³, YU Xiao-Xia³, WU Shan³

1.Department of Pathology|China-Japan Union Hospital|Jilin University| Changchun 130033|China；2.Department of Pathology,Second Hospital|Jilin UniversityChangchun 130041|China；3. Key Laboratory of Pathobiology|Ministry of Educations|School of Basic Medical Science|Jilin University|Changchun 130021|China

Received:2009-01-08 Online:2009-09-28 Published:2009-09-28

摘要/Abstract

摘要：

目的：观察人端粒酶逆转录酶(hTERT)基因转染U2OS细胞对其基质金属蛋白酶2(MMP-2)的影响,探讨hTERT基因与端粒酶的关系及端粒酶活性在肿瘤浸润和转移中的作用。方法：利用重组质粒PCI-Neo-hTERT转染人骨肉瘤细胞系U2OS，选取转染的2个阳性克隆株进行实验，同时以未转染的U2OS细胞为对照。通过RT-PCR和TRAP-ELISA法检测阳性克隆株hTERT基因的转染效果，明胶酶谱法和RT-PCR检测转染hTERT基因对细胞MMP-2的表达和活性的影响。结果：与对照组比较，2个阳性克隆株可见特异性hTERT表达，并表现出明显的端粒酶活性状态（ΔA>0.2　U）。阳性克隆株MMP-2 mRNA的表达无明显改变（P>0.05）,而酶原型MMP-2减少（P<0.01）。结论：hTERT稳定转染克隆U2OS细胞株中异位表达hTERT可以产生端粒酶活性，而且端粒酶活性的出现，可能通过调节MMP的分泌影响细胞外基质成分的降解和构建而对肿瘤的浸润和转移产生一定影响。

关键词: hTERT基因；U2OS细胞；端粒酶活性；基质金属蛋白酶

Abstract:

Abstract:Objective To observe the expression of matrix metalloproteinase-2 (MMP-2) in U2OS cells transfected with human telomerase reverse transcriptase (hTERT),and explore the dependability between and elomerase,and study the effect of telomerase activation in tumor infiltration and metastasis and their possible mechanisms of action. Methods Recombinant plasmid PCI-Neo-hTERT was transfected into human osteosarcoma cell line U2OS.Two transfected positive clonings were chosen for experiment,while normal U2OS cells were used as control group.The effect of hTERT transfected into U2OS cells was detected by RT-PCR and TRAP-ELISA assays,zymography and RT-PCR were used to observe the expression and activity of MMP-2.Results Compared with control group,two positive clonings ectopic generated hTERT mRNA and expressed telomerase activity successfully(ΔA>0.2 U),while the MMP-2 mRNA without obvious difference(P>0.05),and the expression of proMMP-2 was decreased obviously(P<0.01).Conclusion Human osteosarcoma cell line U2OS transfected with hTERT could generate telomerase activity,moreover，the appearance of telomerase activity could regulate the secretion of MMP to affect the degradation and construction of ECM which can affect tumor infiltration and metastasis.

Key words: human telomerase reverse transcriptase gene；U2OS cells；telomerase activity;matrix metalloproteinases

中图分类号:

Q785

姜成威, 杨媚, 石英爱, 于晓霞, 吴珊. 人端粒酶逆转录酶基因转染U2OS细胞对其基质金属蛋白酶2活性的影响[J]. J4, 2009, 35(5): 812-816.

JIANG Cheng-Wei, YANG Mei, DAN Yang-Ai, YU Xiao-Xia, WU Shan. Effect of |U2OS cells transfected with hTERT on activity of matrix metalloproteinase-2[J]. J4, 2009, 35(5): 812-816.