Tmub1对再生肝细胞有丝分裂的影响及其机制

发布时间：2019-02-09 19:25

【摘要】：目的探讨Tmub1对再生肝细胞有丝分裂的影响及与securin的相互作用机制。方法使用40只雌性SD大鼠建立肝大部分切除术后肝再生模型,于术后0、6、12、24、48、72、96、168 h分别取再生肝组织,RT-qPCR和Western blot检测Tmub1和securin在肝再生过程中的mRNA与蛋白表达变化趋势。在大鼠正常肝细胞系BRL-3A中分别建立慢病毒稳定过表达Tmub1、干扰Tmub1和阴性对照组细胞株,用诺考达唑处理获取同步于有丝分裂期的细胞,免疫荧光观察Tmub1过表达组和阴性对照组细胞有丝分裂情况,并用免疫沉淀和Western blot检测Tmub1过表达、Tmub1干扰和阴性对照组BRL-3A细胞中securin的泛素化水平。结果 Tmub1的mRNA和蛋白水平在肝切除术后24、48 h最高,securin的mRNA水平也在48 h最高,而蛋白水平在48 h最低。Tmub1过表达可影响大鼠肝细胞有丝分裂,且securin泛素化水平显著低于阴性对照组(P0.01);Tmub1干扰组securin泛素化水平显著高于阴性对照组(P0.01)。结论 Tmub1可通过抑制securin的泛素化影响大鼠再生肝细胞的有丝分裂。
[Abstract]:Objective to investigate the effect of Tmub1 on mitosis of regenerated hepatocytes and its interaction with securin. Methods 40 female SD rats were used to establish the model of liver regeneration after partial hepatectomy. The regenerated liver tissues were taken out at 6: 12, 24, 48, 72, 96, 168 hours after operation. The changes of mRNA and protein expression of Tmub1 and securin during liver regeneration were detected by RT-qPCR and Western blot. The stable expression of lentivirus Tmub1, interfering with Tmub1 and negative control cell lines were established in normal rat liver cell line BRL-3A, and the cells synchronized with mitotic phase were obtained by nocodazole treatment. The mitosis of Tmub1 overexpression group and negative control group were observed by immunofluorescence. The overexpression of Tmub1 was detected by immunoprecipitation and Western blot. The Ubiquitin level of securin in BRL-3A cells was detected by Tmub1 interference and negative control. Results the mRNA and protein levels of Tmub1 were highest at 24 h after hepatectomy, the mRNA level of securin was the highest at 48 h, and the protein level was lowest at 48 h. The overexpression of Tmub1 could affect the mitosis of rat hepatocytes. The level of securin ubiquitization was significantly lower than that of negative control group (P0.01). The level of securin ubiquitin in Tmub1 interference group was significantly higher than that in negative control group (P0.01). Conclusion Tmub1 can affect the mitosis of rat regenerated hepatocytes by inhibiting the ubiquification of securin.
【作者单位】： 第三军医大学大坪医院野战外科研究所肝胆外科;
【基金】：国家自然科学基金面上项目(81270523)~~
【分类号】：R657.3

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