Ma Liang, Shen Ya-ying, Zhou Peng, Zhou Jun, Guo Feng
Central Laboratory, the First Affiliated Hospital of Soochow University, Suzhou 215006, China.
Zhonghua Zhong Liu Za Zhi. 2012 Mar;34(3):165-8. doi: 10.3760/cma.j.issn.0253-3766.2012.03.002.
To explore how NF-κB family members regulate maspin expression in prostate cancer cells.
The expression of NF-κB subunits and maspin was detected by Western blot analysis in prostate cancer DU145, PC-3, and LNCaP cell lines. RNA interference was performed to analyze whether RelB- or RelA-deletion affectes cell death as well as the expression of NF-κB subunits and maspin. The impact of RelB-silencing in DU145 cells was investigated by flow cytometry. The regulation of RelB on maspin expression in the prostate cancer PC-3 cells was also examined via stable transfection of RelB expression plasmid.
RelA, p50, RelB, and p52 were constitutively expressed in androgen-independent prostate cancer DU145 and PC-3 cells, while RelB had the highest expression in DU145 cells. Low expression of maspin was detected in LNCaP and DU145 cells, but elevated expression in PC-3 cells. RelB-silencing in DU145 cells by siRNA interference upregulated the endogenous expression of maspin and induced cell apoptosis (13.3±4.2)%. Overexpression of RelB in PC-3 cells inhibited the endogenous expression of maspin. RelA-silecing had no significant influence on the endogenous expression of maspin.
The classical and alternative NF-κB activitions are sustained in androgen-independent prostate cancer cell lines. The expressions of RelB and maspin are inversely correlated in these cancer cells. The expression of RelB negatively regulates the endogenous expression of maspin, then interferes the cell survival. RelA is not involved in the regulation of maspin expression.
探讨核因子κB（NF-κB）家族成员如何调节前列腺癌细胞中组织蛋白酶抑制因子maspin的表达。
采用蛋白质免疫印迹分析检测前列腺癌DU145、PC-3和LNCaP细胞系中NF-κB亚基和maspin的表达。进行RNA干扰以分析RelB或RelA缺失是否影响细胞死亡以及NF-κB亚基和maspin的表达。通过流式细胞术研究RelB沉默对DU145细胞的影响。还通过稳定转染RelB表达质粒检测RelB对前列腺癌PC-3细胞中maspin表达的调节作用。
RelA、p50、RelB和p52在雄激素非依赖性前列腺癌DU145和PC-3细胞中组成性表达，而RelB在DU145细胞中表达最高。在LNCaP和DU145细胞中检测到maspin低表达，而在PC-3细胞中表达升高。通过小干扰RNA（siRNA）干扰使DU145细胞中的RelB沉默上调了maspin的内源性表达并诱导细胞凋亡（13.3±4.2）%。PC-3细胞中RelB过表达抑制了maspin的内源性表达。RelA沉默对maspin的内源性表达无显著影响。
经典和替代NF-κB激活在雄激素非依赖性前列腺癌细胞系中持续存在。在这些癌细胞中，RelB和maspin的表达呈负相关。RelB的表达负向调节maspin的内源性表达，进而干扰细胞存活。RelA不参与maspin表达的调节。
