南方医科大学学报

南方医科大学学报 ›› 2021, Vol. 41 ›› Issue (12): 1787-1794.doi: 10.12122/j.issn.1673-4254.2021.12.05

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敲低EEFSEC可体外抑制前列腺癌细胞的增殖、迁移和侵袭

徐彬兵，郝敬兰，谢倩文，萨娜尔，王诗虞，杜晓文，卢海成，高 平，石光耀，董小明   

  1. 陕西师范大学生命科学学院，陕西 西安 710119；山东第一医科大学附属成武医院泌尿外科，山东 菏泽 274200
  • 出版日期:2021-12-20 发布日期:2022-01-05

EEFSEC knockdown inhibits proliferation, migration and invasion of prostate cancer cells in vitro

XU Binbing, HAO Jinglan, XIE Qianwen, SA Naer, WANG Shiyu, DU Xiaowen, LU Haicheng, GAO Ping, SHI Guangyao, DONG Xiaoming   

  1. College of Life Sciences, Shaanxi Normal University, Xian 710119, China; Department of Urology, Chengwu Hospital Affiliated to Shandong First Medical University, Heze 274200, China
  • Online:2021-12-20 Published:2022-01-05

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摘要： 目的 探讨硒代半胱氨酸tRNA特异性真核延伸因子（EEFSEC）对人前列腺癌细胞增殖、迁移和侵袭的影响。方法 采用qRT-PCR法检测人正常前列腺细胞系RWPE1和人前列腺癌细胞系22Rv1、LNcap、Vcap、PC-3细胞中EEFSEC mRNA的表达；收集前列腺癌患者的癌组织和癌旁组织，采用Western blot法检测前列腺癌组织中EEFSEC的蛋白表达情况。慢病毒感染22Rv1 细胞，Western blot检测敲低效率；XTT实验检测各组细胞的增殖能力；划痕实验和Transwell实验用来检测细胞迁移能力；Transwell试验检测细胞侵袭能力；流式细胞术检测细胞周期；qRT-PCR检测敲低EEFSEC后周期相关基因的mRNA水平的变化。结果 与对照组相比，EEFSEC在前列腺癌中高表达（P<0.05）；且EEFSEC高表达导致前列腺癌患者不良预后；感染敲低EEFSEC的慢病毒后，可明显降低22Rv1细胞中EEFSEC的蛋白表达水平；与对照组相比，敲低EEFSEC可显著抑制22Rv1细胞的增殖（P<0.001），迁移（P<0.001）和侵袭能力（P<0.001）；敲低EEFSEC细胞周期主要阻滞在G0/G1期，qRT-PCR实验显示敲低EEFSEC可明显下调C-myc和CCNB1的表达，上调p15的表达。结论 敲低EEFSEC可能通过降低C-myc表达来抑制前列腺癌细胞22Rv1的增殖、迁移和侵袭能力。

关键词: 前列腺癌；EEFSEC；增殖；迁移；侵袭

Abstract: Objective To investigate the role of selenocysteine-tRNA specific eukaryotic elongation factor (EEFSEC) in regulating the proliferation, migration, and invasion of human prostate cancer 22Rv1 cells. Methods We detected EEFSEC mRNA expression levels in human normal prostate cell line RWPE1 and human prostate cancer cell lines 22Rv1, LNCaP, Vcap and PC-3 using qRT-PCR and EEFSEC protein expression in surgical specimens of prostate cancer and adjacent tissues using Western blotting. 22Rv1 cells were infected with a lentiviral vector carrying EEFSEC shRNA or a control lentivirus and the interference efficiency was determined using Western blotting. XTT assay was used to assess the changes in the viability of the infected cells, and Transwell chamber assay was used to examine the changes in cell migration and invasion. The effect of EEFSEC knockdown on cell cycle progression was determined with flow cytometry and by detecting the expressions of cell cycle proteins using qRT-PCR. Results EEFSEC was significantly upregulated in prostate cancer cells (P<0.05), and a high expression of EEFSEC was associated with a poor prognosis of the patients with prostate cancer. In 22Rv1 cells, EEFSEC knockdown significantly suppressed the proliferation (P<0.001), migration (P<0.001) and invasion (P<0.001) of the cells, resulted in cell cycle arrest in G0/G1 phase, obviously inhibited the expression of C-myc and CCNB1, and significantly increased the expression of p15. Conclusion EEFSEC knockdown can inhibit the proliferation, migration, and invasion of prostate cancer cells in vitro possibly by down-regulating the expression of C-myc.

Key words: prostate cancer; EEFSEC; proliferation; migration; invasion