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目的 研究P2嘌呤受体激动剂ATP对不同转移性的人前列腺癌细胞亚系 1E8和 2B4的p38和c Jun氨基末端激酶 (JNK)信号传导途径的影响。方法 以常规的蛋白免疫印迹法 ,用特异识别双磷酸化p38和JNK的特异性抗体 ,检测活化的 (磷酸化的 )p38和JNK。结果 外源性ATP以时间和剂量依赖性的方式激活细胞内的p38,并且在高转移性的 1E8细胞中ATP诱导的p38活化水平明显高于不转移的 2B4细胞。但是在ATP的作用下JNK未见明显激活。P2嘌呤受体拮抗剂苏拉明(suramin)和p38抑制剂SB 2 0 35 80均可有效抑制ATP对p38的激活 ,抑制率分别为 ,1E883%和 79% ,2B481%和 6 9%。两细胞亚系中 ,G蛋白调节剂百日咳毒素均未明显影响ATP对p38的激活。结论 细胞外ATP在恶性肿瘤细胞中能够诱导激活p38信号通路 ,且p38激活水平在转移性不同的前列腺癌细胞亚系间存在差异。我们的研究为恶性肿瘤细胞生长及转移机制研究提供了有效线索。
Objective To investigate the effects of P2 purinoceptor agonist ATP on p38 and c-Jun N-terminal kinase (JNK) signaling pathways in different metastatic human prostate cancer cell lines 1E8 and 2B4. Methods Activated (phosphorylated) p38 and JNK were detected by conventional Western blotting using specific antibodies that specifically recognize both phospho-p38 and JNK. Results Exogenous ATP activates intracellular p38 in a time- and dose-dependent manner, and the level of ATP-induced p38 activation in highly metastatic 1E8 cells is significantly higher than that of non-metastatic 2B4 cells. However, no obvious activation of JNK was observed under the action of ATP. P2 purinergic receptor antagonist suramin and p38 inhibitor SB 2 0 35 80 could effectively inhibit the activation of p38 by ATP, the inhibitory rates were 1E883% and 79%, 2B481% and 69%, respectively. In both cell lines, pertussis toxin, a G protein modulator, did not significantly affect ATP activation of p38. Conclusion Extracellular ATP can induce the activation of p38 signal pathway in malignant tumor cells, and the level of p38 activation is different among subtypes of metastatic prostate cancer cells. Our study provides clues for the study of the mechanism of malignant tumor cell growth and metastasis.