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驱动蛋白家族成员3B(KIF3b)在精原细胞瘤细胞周期中的新作用。

A novel role of KIF3b in the seminoma cell cycle.

作者信息

Shen Hao-Qing, Xiao Yu-Xi, She Zhen-Yu, Tan Fu-Qing, Yang Wan-Xi

机构信息

The Sperm Laboratory, College of Life Sciences, Zhejiang University, 866 Yu Hang Tang Road, Hangzhou 310058, China.

The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou 310003, China.

出版信息

Exp Cell Res. 2017 Mar 1;352(1):95-103. doi: 10.1016/j.yexcr.2017.01.023. Epub 2017 Feb 1.

Abstract

KIF3b is a protein of the kinesin-2 family which plays an important role in intraflagellar transport. Testis cancer is a common cancer among young men. Its diagnostic rate is increasing and over half of the cases are seminomas. Many aspects of the mechanism and gene expression background of this cancer remain unclear. Using western-blotting and semi-quantitative PCR we found high protein levels of KIF3b enrichment in seminoma tissue despite the mRNA levels remaining equivalent to that of normal testicular tissues. The distribution of KIF3b was mainly in cells with division potential. Wound-healing assays and cell counting kit assays showed that the knockdown of KIF3b significantly suppressed cell migration ability, viability and number in HeLa cells. Immunofluorescence images during the cell cycle revealed that KIF3b tended to gather at the spindles and was enriched at the central spindle. This indicated that KIF3b may also have direct impacts upon spindle formation and cytokinesis. By counting the numbers of nuclei, spindles and cells, we found that the rates of multipolar division and multi-nucleation were raised in KIF3b-knockdown cells. In this way we demonstrate that KIF3b functions importantly in mitosis and may be essential to seminoma cell division and proliferation as well as being necessary for normal cell division.

摘要

驱动蛋白相互作用因子3B(KIF3b)是驱动蛋白-2家族的一种蛋白质,在鞭毛内运输中起重要作用。睾丸癌是年轻男性中常见的癌症。其诊断率在上升,超过一半的病例为精原细胞瘤。这种癌症的机制和基因表达背景的许多方面仍不清楚。通过蛋白质免疫印迹法和半定量聚合酶链反应,我们发现尽管精原细胞瘤组织中KIF3b的mRNA水平与正常睾丸组织相当,但KIF3b的蛋白质水平却高度富集。KIF3b的分布主要在具有分裂潜能的细胞中。伤口愈合试验和细胞计数试剂盒试验表明,敲低KIF3b可显著抑制HeLa细胞的迁移能力、活力和数量。细胞周期中的免疫荧光图像显示,KIF3b倾向于聚集在纺锤体上,并在中央纺锤体处富集。这表明KIF3b可能也对纺锤体形成和胞质分裂有直接影响。通过计算细胞核、纺锤体和细胞的数量,我们发现敲低KIF3b的细胞中多极分裂和多核形成的比率增加。通过这种方式,我们证明KIF3b在有丝分裂中起重要作用,可能对精原细胞瘤细胞分裂和增殖至关重要,对正常细胞分裂也必不可少。

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