Overexpression of hTERT extends replicative capacity of human nucleus pulposus cells, and protects against serum starvation-induced apoptosis and cell cycle arrest
Overexpression of hTERT extends replicative capacity of human nucleus pulposus cells, and protects against serum starvation-induced apoptosis and cell cycle arrest
复制标题
DOI：
10.1002/jcb.24082
复制
发表时间：
2012-06-01
影响因子：
4
通讯作者：
Xu, Jiake
中科院分区：
文献类型：
--
作者：
Liang, Weiguo;Ye, Dongping;Xu, Jiake
髓核（NP）细胞是软骨细胞样细胞，其通过合成胶原纤维和蛋白聚糖聚集蛋白聚糖以及产生流体静力学膨胀压力来抵抗压缩负荷，因此在椎间盘中发挥重要作用。在这里，我们报告的生产和永生化的人NP细胞系的正常NP细胞使用稳定转染的重组人端粒酶逆转录酶（hTERT）基因的特性。hTERT转染的NP细胞表现出典型的天然细胞的形态特征。与第一代正常NP细胞相比，hTERT转染的NP细胞生长更快，IGF-1和TGF-β基因表达水平增加。它们成功地传代超过20代，II型胶原蛋白和蛋白聚糖聚集蛋白聚糖表达水平没有显著变化。此外，他们表现出抵抗血清饥饿诱导的细胞凋亡，G1期细胞周期停滞，和基因表达的p53，CCNE 1，Fas和Caspase 3。此外，组织学显示，过表达hTERT的NP细胞植入裸鼠体内后未观察到致瘤性。总之，建立了永生化的人NP细胞系，其具有延长的寿命，保留了与原代亲本NP细胞相似的表型特征，并且应该为研究NP细胞的生物学提供合适的模型。J.细胞。113：21122121，2012。(C)2012 Wiley Periodicals，Inc.
The nucleus pulposus (NP) cells are chondrocyte-like cells that are required for the resistance of compressive loads through the synthesis of collagen fibrils and proteoglycan aggrecans, and the generation of a hydrostatic swelling pressure, and thus play an important role in the intervertebral disc. Here, we report the production and characterization of an immortalized human NP cell line from normal NP cells using stable transfection of recombinant human telomerase reverse transcriptase (hTERT) gene. The hTERT-transfected NP cells exhibited morphological characteristics typical of native cells. When compared with the first generation of normal NP cells, the hTERT-transfected NP cells grew faster and had an increased level of IGF-1 and TGF-beta gene expression. They were successfully passaged over 20 generations without significant change in the levels of type II collagen and proteoglycan aggrecan expression. In addition, they showed resistance to serum starvation-induced apoptosis, G1 cell cycle arrest, and gene expression of p53, CCNE1, Fas, and Caspase 3. Moreover, histology revealed that no tumorigenicity of NP cells over expressing hTERT was observed after they were implanted in nude mice. Taken together, an immortalized human NP cell line was established, which has an extended lifespan, retains phenotypic features similar to primary parent NP cells, and should provide a suitable model for studying the biology of NP cells. J. Cell. Biochem. 113: 21122121, 2012. (C) 2012 Wiley Periodicals, Inc.