[1]马德宾,沈雁兵,王保林,等.Tmub1沉默对大鼠肝部分切除术后肝细胞增殖的影响[J].第三军医大学学报,2013,35(09):850-853.
 Ma Debin,Shen Yanbing,Wang Baolin,et al.Influence of Tmub1 silencing on proliferation of rat hepatocytes after partial hepatectomy[J].J Third Mil Med Univ,2013,35(09):850-853.
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
35卷
期数:
2013年第09期
页码:
850-853
栏目:
论著
出版日期:
2013-05-15

文章信息/Info

Title:
Influence of Tmub1 silencing on proliferation of rat hepatocytes after partial hepatectomy
作者:
马德宾沈雁兵王保林陈平
第三军医大学大坪医院野战外科研究所肝胆外科;中国人民解放军第546医院外科
Author(s):
Ma Debin Shen Yanbing Wang Baolin Chen Ping
Department of Hepatobiliary Surgery, Institute of Surgery Research, Daping Hospital, Third Military Medical University, Chongqing, 400042; Department of Surgery, No. 546 Hospital of PLA, Malan, Xinjiang Uygur Autonomous Region, 841700, China
关键词:
Tmub1肝再生securin慢病毒体内转染
Keywords:
Tmub1 liver regeneration securin lentivirus transfection in vivo
分类号:
R329.28; R394.2; R657.3
文献标志码:
A
摘要:
目的   探讨Tmub1蛋白在肝再生过程中的作用及其在肝再生过程中对Securin蛋白的影响。   方法   54只大鼠随机分为3组即Tmub1 RNAi慢病毒组、空载慢病毒组及正常对照组,其中Tmub1 RNAi慢病毒组、空载慢病毒组注射相应病毒颗粒(3×107 TU),对照组未行注射,2 d后行肝部分切除术(partial hepatectomy, PH),每组分为6个亚组:PH术后0、2、6、12、24、48 h,每亚组3只大鼠,术后提取原代肝细胞、留取肝脏组织标本,利用Real-time PCR和Western blot检测慢病毒干扰效果及Tmub1沉默后对Securin蛋白的影响。MTT实验、流式细胞仪检测原代肝细胞的增殖情况。   结果   Real-time PCR和Western blot表明实验组Tmub1 mRNA和蛋白被有效抑制;Tmub1沉默后securin mRNA表达无明显变化,而G2/M期securin蛋白量明显减少。MTT实验表明Tmub1沉默可明显上调PH术后6~24 h肝细胞的增殖速率;流式细胞分析结果显示实验组处于G2/M期的肝细胞比例明显增高。   结论   Tmub1蛋白在肝再生过程中对肝细胞增殖进程发挥负向调控作用,而该作用可能与其在蛋白质水平影响了Securin 的表达密切相关。
Abstract:
Objective    To investigate the role of transmembrane and ubiquitin-like domain-containing protein 1 (Tmub1) protein in the liver regeneration and its impact on the expression of securin in the process.    Methods    A total of 54 adult male SD rats were randomly divided into Tmub1 RNAi lentivirus group, blank lenitiviral vector group, and control, and they received an injection of lentivirus (3×107TU) or PBS through ileocaecal vein. In 2 d later, partial hepatectomy (PH) were carried out in all rats. Then the liver tissue was collected in 0, 2, 6, 12, 24 and 48 h after model establishment to extract primary hepatocytes and preserve liver tissue. Effect of Tmub1 RNAi lentivirus interference on rat primary hepatocytes was detected by real-time PCR and Western blotting, respectively. Effect of Tmub1 RNAi lentivirus on the growth and cell cycle in rat primary hepatocytes was detected by MTT assay and flow cytometry, respectively.    Results    Real-time PCR and Western blotting showed that the expression of Tmub1 at mRNA and protein levels was significantly inhibited, and the expression of securin at mRNA level had no change, but the expression of securin at protein level was significantly down-regulated. MTT assay showed that Tmub1 gene silencing significantly improved the proliferation in liver cells in 6 to 24 h after PH. Flow cytometry displayed that the liver cells were arrested in G2/M phase.    Conclusion   Tmub1 protein plays a negative role in the process of liver cells’ proliferation, which may be closely related to its effect on the expression of securin at protein level.

参考文献/References:

马德宾, 沈雁兵, 王保林, 等. Tmub1沉默对大鼠肝部分切除术后肝细胞增殖的影响[J].第三军医大学学报,2013,35(9):850-853.

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更新日期/Last Update: 2013-05-03