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日本七鳃鳗的高迁移率族盒转录因子1(HBP1)影响细胞周期调控。

High mobility group box transcription factor 1 (HBP1) from Lampetra japonica affects cell cycle regulation.

作者信息

Song Xiaoping, Gao Xingxing, Lu Jiali, Liang Hongfang, Su Peng, Li Qingwei, Pang Yue

机构信息

College of Life Science, Liaoning Normal University, Dalian, China.

Respiratory Medicine, Affiliated Zhong shan Hospital of Dalian University, Dalian, China.

出版信息

Dev Growth Differ. 2018 Apr;60(3):146-157. doi: 10.1111/dgd.12426. Epub 2018 Mar 8.

DOI:10.1111/dgd.12426
PMID:29520767
Abstract

High mobility group (HMG) box-containing protein 1 (HBP1) is a member of the HMG family of chromosomal proteins. Previous studies have shown that human HBP1 exhibits tumor-suppressor activity. Here, we identified a homologue of HBP1, L-hbp1, in Lampetra japonica. The L-hbp1 gene shared high sequence similarity with its homologues in jawed vertebrates, as shown by bioinformatics analyses. L-hbp1 contains a 1,584-bp open reading frame that encodes 527 amino acids. A pAdenox-L-HBP1 plasmid was constructed and transfected successfully in Raji cells, as revealed by real-time PCR. The overexpression of L-HBP1 reduced cell growth rates, inhibited G1 phase progression, decreased cyclin D1 and c-Myc protein expression, and increased p53 protein expression. Western blot and immunohistochemical assays showed that L-HBP1 was primarily distributed in the heart, kidney, gill and liver of lamprey. Cell cycle analysis revealed that decreased L-HBP1 expression in HBP1 morpholino oligonucleotide-transfected lamprey cells resulted in a decreased fraction of cells in the G1 phase and corresponding increases in the S and G2/M phases. Additionally, treatment of lamprey cardiac cells with pharmacological inhibitors of p38 MAP kinase released the cells from G1 arrest. Together, these results indicated that HBP1 expression in lamprey was correlated with the onset of mitotic arrest in these cells, which have implications for cell cycle regulation.

摘要

高迁移率族(HMG)盒蛋白1(HBP1)是染色体蛋白HMG家族的成员。先前的研究表明,人类HBP1具有肿瘤抑制活性。在此,我们在日本七鳃鳗中鉴定出HBP1的一个同源物,即L-hbp1。生物信息学分析表明,L-hbp1基因与其在有颌脊椎动物中的同源物具有高度的序列相似性。L-hbp1包含一个1584 bp的开放阅读框,编码527个氨基酸。实时PCR显示,构建的pAdenox-L-HBP1质粒成功转染到Raji细胞中。L-HBP1的过表达降低了细胞生长速率,抑制了G1期进程,降低了细胞周期蛋白D1和c-Myc蛋白的表达,并增加了p53蛋白的表达。蛋白质免疫印迹和免疫组织化学分析表明,L-HBP1主要分布在七鳃鳗的心脏、肾脏、鳃和肝脏中。细胞周期分析显示,在转染了HBP1吗啉代寡核苷酸的七鳃鳗细胞中,L-HBP1表达降低导致G1期细胞比例下降,S期和G2/M期细胞比例相应增加。此外,用p38丝裂原活化蛋白激酶的药理学抑制剂处理七鳃鳗心脏细胞可使细胞从G1期阻滞中释放出来。这些结果共同表明,七鳃鳗中HBP1的表达与这些细胞有丝分裂阻滞的发生相关,这对细胞周期调控具有重要意义。

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