[1]宋春丽,任吉华,张祯祯,等.SIRT1基因沉默诱导肝癌细胞老化及其机制[J].第三军医大学学报,2012,34(19):1929-1932.
 Song Chunli,Ren Jihua,Zhang Zhenzhen,et al.Silent information regulator 1 gene induces aging of hepatocellular carcinoma cells via p53/p21 pathway[J].J Third Mil Med Univ,2012,34(19):1929-1932.
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SIRT1基因沉默诱导肝癌细胞老化及其机制(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
34卷
期数:
2012年第19期
页码:
1929-1932
栏目:
论著
出版日期:
2012-10-15

文章信息/Info

Title:
Silent information regulator 1 gene induces aging of hepatocellular carcinoma cells via p53/p21 pathway
作者:
宋春丽任吉华张祯祯陶颖陈娟
重庆医科大学感染性疾病分子生物学教育部重点实验室;重庆医科大学附属儿童医院感染科
Author(s):
Song ChunliRen JihuaZhang ZhenzhenTao YingChen Juan
Key Laboratory of Molecular Biology on Infectious Diseases of Ministry of Education, Chongqing Medical University, Chongqing, 400016; Department of Infectious Diseases, Children’s Hospital, Chongqing Medical University, Chongqing, 400014, China
关键词:
沉默信息调节因子1肝细胞癌肿瘤细胞系细胞衰老
Keywords:
silent information regulator 1 hepatocellular carcinoma tumor cell linecell aging
分类号:
R394.2; R73-354; R735.7
文献标志码:
A
摘要:
目的      探讨慢病毒介导的沉默信息调节因子1(silent information regulator1,SIRT1)基因沉默对肝癌细胞老化的影响及机制。      方法       通过慢病毒介导的shRNA干扰技术靶向沉默SIRT1的表达,并通过Real-time PCR和Western blot验证SIRT1基因的沉默效果。BrdU标记实验检测细胞增殖情况,流式细胞仪检测细胞周期变化;β-半乳糖苷酶染色观察肝癌细胞有无老化;Western blot检测衰老相关基因p53和p21蛋白的变化。      结果       慢病毒介导的shRNA能显著抑制细胞SIRT1的mRNA及蛋白水平。SIRT1沉默能抑制肝癌细胞的增殖,抑制率达70%~80%;同时,SIRT1沉默能显著诱导G1期细胞周期阻滞:感染shCont的细胞处于G1期比例为(44.29±0.36)%,而感染shSIRT1-1的细胞处于G1期细胞为(69.20±1.24)%,感染shSIRT1-2的细胞处于G1期比例为(65.86±1.75)%。SIRT1沉默后肝癌细胞中衰老相关的β-半乳糖苷酶染色显著增高,阳性细胞占40%~50%(P<0.05),并伴随衰老相关基因p53和p21蛋白表达增加。      结论       SIRT1基因沉默可能通过p53/p21途径促进细胞衰老。
Abstract:
Objective      To study the effect of lentivirus-mediated silent information regulator 1 gene (SIRT1) on aging of hepatocellular carcinoma cells (HCCs) and its mechanism.       Methods      SIRT1 expression was targeted by lentivirus-mediated shRNA interference technology. Effect of SIRT1 on aging of HCCs was detected by RT-PCR and Western blot analysis, respectively. Proliferation of HCCs was assayed by BrdU-labeling test.  Cell cycle of HCCs was displayed by flow cytometry. Aging of HCCs was observed with SA-β-gal staining. Aging-related p53 and p21 proteins were demonstrated by Western blot analysis.       Results      Lentivirus-mediated shRNA significantly down-regulated the SIRT1 mRNA and protein expression levels in HCCs. SIRT1 suppressed the proliferation of HCCs with a suppression rate of 70%-80%, and significantly induced cell cycle arrest at stage G1. The proportion of shCont-, SIRT1- and SIRT2-infected HCCs at stage G1 was about (42.29±0.36)%, (69.20±1.24)%, and (65.86±1.75)%, respectively. The aging-related β-galactosidase-stained HCCs were significantly greater with a positive rate of 40%-50% (P<0.05), and the aging-related p53 and p21 protein expression levels were significantly higher.       Conclusion      SIRT1 promotes the aging of HCCs possibly through the p53/p21 pathway.

参考文献/References:

宋春丽, 任吉华, 张祯祯, 等. SIRT1基因沉默诱导肝癌细胞老化及其机制[J]. 第三军医大学学报,2012,34(19):1929-1932.

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更新日期/Last Update: 2012-09-27