蛋白激酶D3参与前列腺癌细胞中基质金属蛋白酶-7的负调控

[Protein kinase D3 is involved in negative regulation of MMP-7 in prostate cancer cells].

作者信息

Zou Zhi-peng, Feng Li, Xu Wan-fu, Ke Zhi-yong, Wang Q Jane, Deng Fan

机构信息

Department of Cell Biology, Southern Medical University, Guangzhou 510515, China.

出版信息

Nan Fang Yi Ke Da Xue Xue Bao. 2010 Aug;30(8):1767-70.

PMID:20813660

Abstract

OBJECTIVE

To explore the role of protein kinase D3 (PKD3) in the regulation of matrix metalloproteinases 7 (MMP-7) expression in prostate cancer cells.

METHODS

PC-3 cells were either stimulated with 100 nmol/L PMA to activate PKD3 kinase activity, or transiently transfected with PKD3 siRNA, and the relative expression level of MMP-7 mRNA were analyzed by real-time PCR using 2(-delta delta Ct) method. MMP-7 mRNA levels were also analyzed and quantified in HEK293 cells with over-expression of wild-type PKD3, PKD3 knockdown (using PKD3 siRNA), or over-expression of wild-type PKD3 followed by PKD3 knockdown.

RESULTS

MMP-7 mRNA expression in PC3 cells was significantly decreased after PMA-induced PKD3 kinase activation. In contrast, PKD3 knockdown by siRNA transfection markedly increased MMP-7 mRNA level (P<0.01). MMP-7 mRNA level in HEK293 cells was significantly decreased by PKD3 over-expression, whereas obviously increased by PKD3 knockdown. Down-regulation of MMP-7 mRNA level in HEK293 induced by PKD3 over-expression was rescued by PKD3 knockdown.

CONCLUSION

PKD3 may contribute to the malignant progression of prostate cancer cells through negative regulation of MMP-7 expression.

摘要

目的

探讨蛋白激酶D3（PKD3）在前列腺癌细胞中对基质金属蛋白酶7（MMP - 7）表达调控的作用。

方法

用100 nmol/L佛波酯（PMA）刺激PC - 3细胞以激活PKD3激酶活性，或用PKD3小干扰RNA（siRNA）进行瞬时转染，采用2^（-ΔΔCt）法通过实时定量聚合酶链反应（PCR）分析MMP - 7信使核糖核酸（mRNA）的相对表达水平。在过表达野生型PKD3、敲低PKD3（使用PKD3 siRNA）或过表达野生型PKD3后再敲低PKD3的人胚肾293（HEK293）细胞中，也对MMP - 7 mRNA水平进行了分析和定量。

结果

PMA诱导PKD3激酶激活后，PC3细胞中MMP - 7 mRNA表达显著降低。相反，通过siRNA转染敲低PKD3可显著提高MMP - 7 mRNA水平（P<0.01）。PKD3过表达使HEK293细胞中MMP - 7 mRNA水平显著降低，而敲低PKD3则使其明显升高。PKD3敲低可挽救PKD3过表达诱导的HEK293细胞中MMP - 7 mRNA水平的下调。