Xu B, Hao J, Xie Q, Sa N, Wang S, DU X, Lu H, Gao P, Shi G, Dong X
College of Life Sciences, Shaanxi Normal University, Xian 710119, China.
Department of Urology, Chengwu Hospital Affiliated to Shandong First Medical University, Heze 274200, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2021 Dec 20;41(12):1787-1794. doi: 10.12122/j.issn.1673-4254.2021.12.05.
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.
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.
EEFSEC was significantly upregulated in prostate cancer cells ( < 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 ( < 0.001), migration ( < 0.001) and invasion ( < 0.001) of the cells, resulted in cell cycle arrest in G/G phase, obviously inhibited the expression of C-myc and CCNB1, and significantly increased the expression of p15.
EEFSEC knockdown can inhibit the proliferation, migration, and invasion of prostate cancer cells in vitro possibly by down-regulating the expression of C-myc.
探讨硒代半胱氨酸 - tRNA特异性真核延伸因子(EEFSEC)在调节人前列腺癌22Rv1细胞增殖、迁移和侵袭中的作用。
我们使用qRT-PCR检测人正常前列腺细胞系RWPE1和人前列腺癌细胞系22Rv1、LNCaP、Vcap及PC-3中EEFSEC mRNA表达水平,使用蛋白质免疫印迹法检测前列腺癌手术标本及癌旁组织中EEFSEC蛋白表达。用携带EEFSEC shRNA的慢病毒载体或对照慢病毒感染22Rv1细胞,通过蛋白质免疫印迹法测定干扰效率。采用XTT法评估感染细胞活力的变化,采用Transwell小室法检测细胞迁移和侵袭的变化。通过流式细胞术并使用qRT-PCR检测细胞周期蛋白的表达来确定EEFSEC敲低对细胞周期进程的影响。
EEFSEC在前列腺癌细胞中显著上调(<0.05),EEFSEC高表达与前列腺癌患者的不良预后相关。在22Rv1细胞中,EEFSEC敲低显著抑制细胞的增殖(<0.001)、迁移(<0.001)和侵袭(<0.001),导致细胞周期停滞在G/G期,明显抑制C-myc和CCNB1的表达,并显著增加p15的表达。
EEFSEC敲低可能通过下调C-myc的表达在体外抑制前列腺癌细胞的增殖、迁移和侵袭。