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目的探讨丝裂原活化蛋白激酶在17β雌二醇(E2)抑制前列腺癌PC3细胞生长中的作用。方法检测不同浓度E2作用不同时间后PC3细胞生长抑制率。流式细胞仪分析细胞周期分布,TUNEL染色检测凋亡。Western blot检测ERK1/2,JNK和p38活性。RT-PCR法检测雌激素受体(ER)α、ERβ、P21WAF1和cyclinD1的表达。结果E2抑制PC3细胞增殖,并且可以激活ERK1/2、JNK和p38。处理因素作用48h后,对照组、E2、E2+PD98059、E2+SP600125、E2+SB203580组细胞的凋亡率分别为(0.9±0.1)%;(23.0±1.4)%;(30.0±1.2)%;(10.6±0.8)%和(14.6±0.7)%,(P<0.05)。E2使细胞阻滞在G1期,PD98059、SP600125、SB203580分别预处理1h后细胞分别进一步阻滞在G1期;轻度阻滞在G1期和进入S期。RT-PCR发现PC3细胞表达ERα和ERβ,E2、E2+PD98059、E2+SP600125、E2+SB203580组中cyclinD1、P21WAF1基因表达分别为对照组的(0.42±0.03)、(3.13±0.02)倍;(0.21±0.03)、(3.08±0.05)倍;(0.43±0.01)、(1.31±0.04)倍;(2.81±0.02)、(3.14±0.02)倍(P<0.05)。结论E2激活JNK增加P21WAF1基因表达,并激活p38抑制cyclinD1表达,使细胞阻滞在G1期,JNK和p38通路还介导E2引起PC3细胞凋亡。同时E2又可激活ERK1/2,轻度拮抗上述作用。
Objective To investigate the role of mitogen-activated protein kinase in inhibiting the growth of prostate cancer PC3 cells by 17β estradiol (E2). Methods The growth inhibition rates of PC3 cells after different concentrations of E2 were detected. Cell cycle distribution was analyzed by flow cytometry, and apoptosis was detected by TUNEL staining. Western blot detection of ERK1 / 2, JNK and p38 activity. The expression of estrogen receptor (ER) α, ERβ, P21WAF1 and cyclinD1 were detected by RT-PCR. Results E2 inhibited PC3 cell proliferation and could activate ERK1 / 2, JNK and p38. The apoptosis rates of control group, E2, E2 + PD98059, E2 + SP600125 and E2 + SB203580 group were (0.9 ± 0.1)%, (23.0 ± 1.4)% and (30.0 ± 1.2)% ; (10.6 ± 0.8)% and (14.6 ± 0.7)%, respectively (P <0.05). E2 retarded cells in G1 phase. Cells pretreated with PD98059, SP600125 and SB203580 for 1 h, respectively, further arrested in G1 phase and slightly blocked in G1 phase and entered S phase. The expression of cyclinD1 and P21WAF1 in E2, E2, PD98059, E2 + SP600125 and E2 + SB203580 groups was (0.42 ± 0.03), (3.13 ± 0.02) times of the control group, respectively, in PC3 cells by RT- 0.21 ± 0.03), (3.08 ± 0.05) times, (0.43 ± 0.01), (1.31 ± 0.04) times, (2.81 ± 0.02) and (3.14 ± 0.02) times (P <0.05). Conclusion E2 activates JNK to increase the expression of P21WAF1 gene and activate p38 to inhibit the expression of cyclinD1, which blocks cell cycle arrest in G1 phase. JNK and p38 pathway also mediate E2-induced PC3 cell apoptosis. At the same time E2 can activate ERK1 / 2, mild antagonism of the above effects.