Family Planning Research Institute, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
The First People's Hospital of Tianmen City, Tianmen, China.
Cell Prolif. 2019 Mar;52(2):e12549. doi: 10.1111/cpr.12549. Epub 2018 Nov 28.
CSE1L has been reported to be highly expressed in various tumours. Testicular germ cell tumours are common among young males, and seminoma is the major type. However, whether CSE1L has functions in the seminoma is unclear.
The expression of CSE1L was detected by immunohistochemistry in seminoma tissues and non-tumour normal testis tissues from patients. CSE1L distribution during cell mitosis was determined by immunofluorescent staining with CSE1L, α-tubulin and γ-tubulin antibodies. The effects of Cse1L knockdown on cell proliferation and cell cycle progression were determined by Cell Counting Kit-8 assay, flow cytometry, PH3 staining and bromodeoxyuridine incorporation assay.
CSE1L was significantly enriched in the seminoma tissue compared with the non-tumour normal testis tissue. CSE1L also co-localized with α-tubulin in the cells with a potential to divide. In the seminoma cell line TCam-2, CSE1L was associated with the spindles and the centrosomes during cell division. The knockdown of CSE1L in TCam-2 cells attenuated the cells' proliferative capacity. Cell cycle assay revealed that the CSE1L-deficient cells were mainly arrested in the G0/G1 phase and moderately delayed in the G2/M phase. The proportion of cells with multipolar spindle and abnormal spindle geometry was obviously increased by CSE1L expression silencing in the TCam-2 cells.
Overall, these findings showed that CSE1L plays a pivotal role in maintaining cell proliferation and cell division in seminomas.
CSE1L 在多种肿瘤中表达水平较高。睾丸生殖细胞肿瘤在年轻男性中较为常见,其中精原细胞瘤是主要类型。然而,CSE1L 在精原细胞瘤中是否具有功能尚不清楚。
采用免疫组织化学法检测 CSE1L 在精原细胞瘤组织和患者非肿瘤正常睾丸组织中的表达。通过用 CSE1L、α-微管蛋白和γ-微管蛋白抗体进行免疫荧光染色,确定 CSE1L 在细胞有丝分裂过程中的分布。通过细胞计数试剂盒-8 检测、流式细胞术、PH3 染色和溴脱氧尿苷掺入实验,确定 Cse1L 敲低对细胞增殖和细胞周期进程的影响。
CSE1L 在精原细胞瘤组织中明显富集,而非肿瘤正常睾丸组织中表达水平较低。CSE1L 还与有潜在分裂能力的细胞中的α-微管蛋白共定位。在精原细胞瘤细胞系 TCam-2 中,CSE1L 在细胞分裂过程中与纺锤体和中心体相关。在 TCam-2 细胞中敲低 CSE1L 会减弱细胞的增殖能力。细胞周期检测显示,CSE1L 缺陷细胞主要停滞在 G0/G1 期,在 G2/M 期中度延迟。沉默 TCam-2 细胞中的 CSE1L 表达会明显增加多极纺锤体和异常纺锤体形态的细胞比例。
综上所述,这些发现表明 CSE1L 在维持精原细胞瘤细胞增殖和细胞分裂中发挥着关键作用。