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目的 在高转移性的人前列腺癌细胞 1E8中 ,探讨丝裂原活化蛋白激酶磷酸酶 5(MKP5 )在ATP激活丝裂原活化蛋白激酶 (MAPK)信号通路及相关生物学行为中的作用。方法 构建野生型 pcDL SRα MKP5表达载体 ,稳定转染 1E8细胞并筛选阳性克隆。应用免疫印迹法检测细胞ERK1/ 2和 p38的活化情况。应用生长曲线、体外侵袭实验及软琼脂集落形成实验检测ATP刺激下 ,分别应用MEK抑制剂PD980 5 9及p38抑制剂SB2 0 35 80后各组转染细胞的生物学行为。结果野生型MKP5可明显抑制ATP对 p38通路的激活 ,明显逆转ATP长期刺激下对 1E8细胞的生长抑制作用 ,并明显抑制ATP短期刺激下对 1E8细胞体外侵袭能力的促进作用 ,其作用与SB2 0 35 80相似。PD980 5 9可以部分逆转ATP对转染细胞的体外生长及软琼脂集落形成能力的抑制作用 ,但对ATP短期刺激下对体外侵袭能力的促进作用不明显。结论 在ATP刺激的不同时相里 ,p38、ERK1/ 2通路以不同的方式参与调节肿瘤细胞的生长、侵袭等过程 ,MKP5可通过抑制p38通路的方式参与抑制ATP的这些作用。
Objective To investigate the role of mitogen-activated protein kinase phosphatase 5 (MKP5) in ATP-activated mitogen-activated protein kinase (MAPK) signaling pathway and its related biological behavior in highly metastatic human prostate cancer cell line 1E8. Methods The wild-type pcDL SRα MKP5 expression vector was constructed and stably transfected into 1E8 cells and the positive clones were screened. The activation of ERK1 / 2 and p38 was detected by Western blotting. The growth curves, in vitro invasion assay and soft agar colony formation assay were used to detect the biological behavior of transfected cells after being stimulated by ATP, MEK inhibitor PD980 5 9 and p38 inhibitor SB2 0 35 80 respectively. Results Wild-type MKP5 could significantly inhibit the activation of p38 pathway by ATP, reversing the inhibitory effect of ATP on the growth of 1E8 cells under long-term stimulation of ATP, and significantly inhibited the invasive ability of 1E8 cells in vitro under short-term ATP stimulation. 35 80 Similar. PD980 5 9 could partially reverse the inhibitory effect of ATP on the growth of transfected cells in vitro and the ability of colony formation of soft agar. However, PD980 5 9 did not obviously promote the invasive ability in vitro. Conclusions In different phases of ATP stimulation, p38 and ERK1 / 2 pathways are involved in the regulation of tumor cell growth and invasion in different ways. MKP5 may be involved in the inhibition of ATP by inhibiting the p38 pathway.