Posttranscriptional induction of p21Waf1 mediated by ectopic p16INK4 in human diploid fibroblast
Posttranscriptional induction of p21Waf1 mediated by ectopic p16INK4 in human diploid fibroblast
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DOI：
10.1097/00029330-200703010-00011
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发表时间：
2007-03
影响因子：
6.1
通讯作者：
Xiao‐lin Han;Fuyong Wu;Zong-yu Zhang;T. Tong
中科院分区：
文献类型：
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作者：
Xiao‐lin Han;Fuyong Wu;Zong-yu Zhang;T. Tong
背景p16INK4和p21Waf1都是肿瘤抑制因子，在调节细胞衰老方面具有相似的生物学功能。先前的报道表明，在HeLa细胞中，p21Waf1可以通过转录因子Sp1激活p16INK4。本研究旨在确定p16INK4对p21Waf1表达和功能的影响。方法用正义（2BS/p16INK4）、反义p16INK4 （2BS/asp16INK4）或空载体（2BS/neo）稳定转染人二倍体成纤维细胞2BS细胞，采用逆转录聚合酶链反应（RT - PCR）、荧光活化细胞分选（FACS）和Western blot检测。结果2BS/p16INK4细胞在G1期和G2/M期均表现出细胞周期阻滞。内源性p21Waf1蛋白水平在2BS/p16INK4细胞中增加了两倍，但在2BS/asp16INK4细胞中没有下降。p21Waf1 mRNA水平在2BS/p16INK4和2BS/asp16INK4细胞中均未受影响。结论p16INK4可能通过转录后机制调控p21Waf1蛋白水平，在细胞衰老过程中发挥重要作用。
Background Both p16INK4 and p21Waf1 are tumor suppressors with similar biological functions in the regulation of cellular senescence. Previous reports showed that p16INK4 could be activated by p21Waf1 through transcriptional factor Sp1 in HeLa cells. This study was undertaken to determine the effects of p16INK4 on the expression and functions of p21Waf1. Methods Human diploid fibroblast 2BS cells were stably transfected with sense (2BS/p16INK4), antisense p16INK4 (2BS/asp16INK4) or empty vector (2BS/neo) Then they were assayed by reverse‐transcription polymerase chain reaction (RT‐PCR), fluorescence activated cell sorting (FACS) and Western blot. Results 2BS/p16INK4 cells exhibited cell cycle arrest in both G1 and G2/M phases. Endogenous p21Waf1 protein levels increased twofold in the 2BS/p16INK4 cells, but not decreased in the 2BS/asp16INK4 cells. p21Waf1 mRNA levels were not affected in neither 2BS/p16INK4 nor 2BS/asp16INK4 cells. Conclusion p16INK4 may play an important role in the regulation of cellular senescence by modulating the p21Waf1 protein level via the posttranscriptional mechanism.