论文部分内容阅读
目的评估LNCaP细胞中雄激素依赖性和雄激素非依赖性蛋白质组学特征,探讨前列腺癌的雄激素非依赖性生长的分子机制。方法雄激素敏感性LNCaP细胞在常规细胞培养基中持续培养。在雄激素递减环境中培养雄激素敏感的LNCaP细胞,且培养出了稳定传代的细胞,命名为LNCaP-s细胞。检测两种不同的LNCaP细胞生长特性和对雄激素的反应,观察饥饿细胞的形态学变化。通过双相凝胶电泳(2-DE)和基质辅助激光解析电离/飞行时间质谱测量法(MALDI-TOF MS)分析前列腺癌LNCaP细胞的蛋白表达。结果在2-DE LNCaP蛋白谱中,成功鉴定出了222种蛋白质。与LNCaP细胞相比,在LNCaP-s细胞中发现150种蛋白表达差异有统计学意义,成功鉴定出75种蛋白质。LNCaP蛋白谱中发现在LNCaP-s细胞中有26种蛋白表达上调或下调,且其蛋白受生长激素抑素(sms)以及其类似物(smsdx)的影响。鉴定出的蛋白参与多种生理过程,包括应激反应、细胞内信号转导和凋亡。在LNCaP-s细胞中,150种蛋白中7种蛋白表达上调,其余表达下调。蛋白质表达的定性分析显示,LNCaP细胞中有6种蛋白在LNCaP-s细胞中无表达。与LNCaP细胞相比,LNCaP-s细胞系中未发现新蛋白。结论LNCaP细胞中雄激素依赖性和雄激素非依赖性蛋弊质表达不同,且受sms及smsdx影响,可提供潜在的诊断和预后判断的标记物及药物治疗靶点并充实前列腺癌蛋白库,对基础和临床研究有重要意义。
Objective To evaluate the androgen-independent androgen-independent proteomic characteristics of LNCaP cells and to explore the molecular mechanism of androgen-independent growth of prostate cancer. Methods Androgen-sensitive LNCaP cells were cultured continuously in conventional cell culture medium. Androgen-sensitive LNCaP cells were cultured in an androgen-descending environment, and stable passaged cells were cultured and named as LNCaP-s cells. The growth characteristics of different LNCaP cells and the response to androgen were examined to observe the morphological changes of starved cells. Prostate cancer LNCaP cells were analyzed for protein expression by two-phase gel electrophoresis (2-DE) and matrix-assisted laser desorption ionization / time of flight mass spectrometry (MALDI-TOF MS). Results In the 2-DE LNCaP protein profile, 222 proteins were successfully identified. Compared with LNCaP cells, 150 proteins were found to be statistically significant in LNCaP-s cells, and 75 proteins were successfully identified. 26 of the LNCaP-s cells were found to have up-regulated or down-regulated LNCaP protein profiles and their proteins were affected by sms and its analogues (smsdx). The identified proteins are involved in a variety of physiological processes including stress response, intracellular signal transduction and apoptosis. In LNCaP-s cells, seven of the 150 proteins were up-regulated and the rest were down-regulated. Qualitative analysis of protein expression showed that LNCaP cells in six kinds of protein expression in LNCaP-s cells. Compared to LNCaP cells, no new protein was found in the LNCaP-s cell line. Conclusion The expressions of androgen-dependent and androgen-independent proteins in LNCaP cells are different, and are affected by sms and smsdx, which may provide potential diagnostic and prognostic markers and drug therapy targets and enrich the prostate cancer protein library. Basic and clinical research is of great importance.