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目的癌症晚期阶段,随着肿瘤体积增大,肿瘤灶内的细胞面临着低氧、营养匮乏的生存环境。近年来研究表明,癌细胞在这种环境条件下仍具有一定的增殖能力。本研究旨在探讨血清饥饿对前列腺癌(prostate cancer,PCa)DU145细胞增殖的影响及其相关信号分子的调控机制。方法无血清培养基干预DU145细胞,分别用MTT、细胞计数实验观察血清饥饿对细胞活力和细胞增殖的影响;蛋白质印迹法测定干预前后细胞外调节蛋白激酶(extracellular signal-regulated kinase,Erk)、丝氨酸/苏氨酸蛋白激酶(serine/threonine protein kinase,Akt)的蛋白表达及磷酸化水平变化。结果血清饥饿72h,细胞活力(t=8.233,P=0.001)和细胞增殖能力(t=12.291,P=0.000 3)显著降低。血清饥饿48h的细胞数目为0.792±0.063,较血清饥饿24h的0.591±0.070增多,t=3.686,P=0.021。随着血清饥饿时间的延长,细胞增殖受到影响。蛋白质印迹法结果表明,血清饥饿能明显促进Erk(P=0.007)和Akt(P=0.001)磷酸化,却对Erk和Akt总蛋白的表达没有显著影响。Erk通路抑制剂U0126显著抑制了血清饥饿环境下Erk的磷酸化(P<0.001),同时抑制了细胞的增殖能力,t=8.544,P=0.001;Akt通路抑制剂LY294002显著抑制血清饥饿环境下Akt磷酸化(P=0.003)的同时抑制了细胞的增殖能力,t=10.472,P<0.001。结论在血清饥饿条件下,DU145细胞可以通过激活Erk和Akt信号通路来保持一定的增殖能力,这可能是其在血清饥饿环境下做出的适应性反应。
Objective Cancer late stage, with the tumor volume increases, the tumor cells within the tumor face hypoxic, nutrient-deficient living environment. In recent years, studies have shown that cancer cells in this environment still has a certain degree of proliferation. The purpose of this study was to investigate the effects of serum starvation on the proliferation of prostate cancer (PCa) DU145 cells and the related regulatory mechanisms of signal molecules. Methods The serum-free medium was used to intervene in DU145 cells. The effects of serum starvation on cell viability and cell proliferation were observed by MTT and cell counting. The levels of extracellular signal-regulated kinase (Erk), serine / Threonine protein kinase (serine / threonine protein kinase, Akt) protein expression and phosphorylation level changes. Results Serum starvation 72h, cell viability (t = 8.233, P = 0.001) and cell proliferation ability (t = 12.291, P = 0.0003) decreased significantly. The number of cells at 48h after serum starvation was 0.792 ± 0.063, which was significantly higher than 0.591 ± 0.070 at 24h after serum starvation, t = 3.686, P = 0.021. With the prolongation of serum starvation, cell proliferation is affected. Results of Western blotting showed that serum starvation significantly promoted the phosphorylation of Erk (P = 0.007) and Akt (P = 0.001), but had no significant effect on the expression of Erk and Akt proteins. Erk pathway inhibitor U0126 significantly inhibited Erk phosphorylation in serum-starved environment (P <0.001) and inhibited proliferation of cells, t = 8.544, P = 0.001; Akt pathway inhibitor LY294002 significantly inhibited Akt Phosphorylation (P = 0.003) inhibited proliferation of cells at the same time, t = 10.472, P <0.001. Conclusions Under serum starvation conditions, DU145 cells can maintain certain proliferative capacity by activating Erk and Akt signaling pathways, which may be the adaptive response in serum starvation.