【摘要】：目的探討Tmub1對再生肝細胞有絲分裂的影響及與securin的相互作用機制。方法使用40只雌性SD大鼠建立肝大部分切除術(shù)后肝再生模型,于術(shù)后0、6、12、24、48、72、96、168 h分別取再生肝組織,RT-qPCR和Western blot檢測Tmub1和securin在肝再生過程中的mRNA與蛋白表達變化趨勢。在大鼠正常肝細胞系BRL-3A中分別建立慢病毒穩(wěn)定過表達Tmub1、干擾Tmub1和陰性對照組細胞株,用諾考達唑處理獲取同步于有絲分裂期的細胞,免疫熒光觀察Tmub1過表達組和陰性對照組細胞有絲分裂情況,并用免疫沉淀和Western blot檢測Tmub1過表達、Tmub1干擾和陰性對照組BRL-3A細胞中securin的泛素化水平。結(jié)果 Tmub1的mRNA和蛋白水平在肝切除術(shù)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過表達可影響大鼠肝細胞有絲分裂,且securin泛素化水平顯著低于陰性對照組(P0.01);Tmub1干擾組securin泛素化水平顯著高于陰性對照組(P0.01)。結(jié)論 Tmub1可通過抑制securin的泛素化影響大鼠再生肝細胞的有絲分裂。
[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.
【作者單位】： 第三軍醫(yī)大學(xué)大坪醫(yī)院野戰(zhàn)外科研究所肝膽外科;
【基金】：國家自然科學(xué)基金面上項目(81270523)~~
【分類號】：R657.3

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