Lee Ming-Shyue, Igawa Tsukasa, Yuan Ta-Chun, Zhang Xiu-Qing, Lin Fen-Fen, Lin Ming-Fong
Department of Biochemistry and Molecular Biology, University of Nebraska Medical Center, Omaha 68198, USA.
Oncogene. 2003 Feb 6;22(5):781-96. doi: 10.1038/sj.onc.1206066.
The expression and secretion of prostate-specific antigen (PSA) are regulated by androgens in normal prostate secretory epithelial cells. In prostate cancer patients, the serum PSA level is usually elevated and cancer cells are initially responsive to androgens. However, those cancer cells become androgen-independent after androgen ablation therapy. In hormone-refractory cancer patients, even in an androgen-deprived environment, the circulation level of PSA rebounds and is constitutively elevated through a yet unknown mechanism. Tyrosine phosphorylation of ErbB-2 is involved in regulating the androgen-responsive phenotype of prostate cancer cells, and it is at least partly regulated by the cellular form of prostatic acid phosphatase (PAcP), a prostate-unique protein tyrosine phosphatase. We investigated the ErbB-2 signal pathway in androgen-independent PSA secretion. LNCaP C-81 cells, which are androgen-independent LNCaP cells lacking endogenous PAcP expression with a hypertyrosine phosphorylated ErbB-2, secreted a higher level of PSA in conditioned media than did androgen-sensitive LNCaP C-33 parental cells. A restored expression of cellular PAcP in C-81 cells was concurrent with a decrease in tyrophosphorylation of ErbB-2 and reduction of PSA secretion. Moreover, transient transfection of C-33 cells with the wild-type ErbB-2 or a constitutively active mutant of MEK1 cDNA resulted in an increased level of secreted PSA. The elevation of secreted PSA level by the forced expression of ErbB-2 was inhibited by an MEK inhibitor, PD98059. In C-81 cells, the expression of a dominant negative mutant of ErbB-2 reduced the secreted level of PSA. The inhibition of ErbB-2 or mitogen-activated protein (MAP) kinases by specific inhibitors AG879, AG825, or PD98059 led to a decrease in PSA secretion. Taken together, our data clearly indicate that the ErbB-2 signal pathway via MAP kinases (ERK1/2) is involved in regulating the secretion of PSA by androgen-independent human prostate cancer LNCaP C-81 cells in an androgen-depleted environment.
在正常前列腺分泌上皮细胞中,前列腺特异性抗原(PSA)的表达和分泌受雄激素调节。在前列腺癌患者中,血清PSA水平通常会升高,且癌细胞最初对雄激素有反应。然而,在雄激素去除治疗后,这些癌细胞会变得对雄激素不依赖。在激素难治性癌症患者中,即使在雄激素缺乏的环境中,PSA的循环水平也会反弹,并通过一种未知机制持续升高。ErbB-2的酪氨酸磷酸化参与调节前列腺癌细胞的雄激素反应表型,并且它至少部分受前列腺特异性蛋白酪氨酸磷酸酶——前列腺酸性磷酸酶(PAcP)的细胞形式调节。我们研究了雄激素非依赖性PSA分泌中的ErbB-2信号通路。LNCaP C-81细胞是缺乏内源性PAcP表达且具有高酪氨酸磷酸化ErbB-2的雄激素非依赖性LNCaP细胞,与雄激素敏感的LNCaP C-33亲本细胞相比,其在条件培养基中分泌的PSA水平更高。C-81细胞中细胞PAcP的恢复表达与ErbB-2的酪氨酸磷酸化减少及PSA分泌降低同时发生。此外,用野生型ErbB-2或MEK1 cDNA的组成型活性突变体瞬时转染C-33细胞会导致分泌的PSA水平升高。MEK抑制剂PD98059可抑制因ErbB-2的强制表达而导致的分泌PSA水平升高。在C-81细胞中,ErbB-2显性负性突变体的表达降低了PSA分泌水平。特异性抑制剂AG879、AG825或PD98059对ErbB-2或丝裂原活化蛋白(MAP)激酶的抑制导致PSA分泌减少。综上所述,我们的数据清楚地表明,在雄激素缺乏的环境中,通过MAP激酶(ERK1/2)的ErbB-2信号通路参与调节雄激素非依赖性人前列腺癌LNCaP C-81细胞中PSA的分泌。